Engineering of hydroxymandelate synthases and the aromatic amino acid pathway enables de novo biosynthesis of mandelic and 4-hydroxymandelic acid with Saccharomyces cerevisiae
Mandelic acid (MA) and 4-hydroxymandelic acid (HMA) are valuable specialty chemicals used as precursors for flavors as well as for cosmetic and pharmaceutical purposes. Today they are mainly synthesized chemically. Their synthesis through microbial fermentation would allow for environmentally sustai...
Ausführliche Beschreibung
Gespeichert in:
| Autor*in: |
Reifenrath, Mara [verfasserIn] |
|---|
| Format: |
E-Artikel |
|---|---|
| Sprache: |
Englisch |
| Erschienen: |
2018transfer abstract |
|---|
| Schlagwörter: |
|---|
| Umfang: |
9 |
|---|
| Übergeordnetes Werk: |
Enthalten in: Inhibition on polysulfides dissolve during the discharge-charge by using fish-scale-based porous carbon for lithium-sulfur battery - Gao, Mengyao ELSEVIER, 2014transfer abstract, Orlando, Fla |
|---|---|
| Übergeordnetes Werk: |
volume:45 ; year:2018 ; pages:246-254 ; extent:9 |
| Links: |
|---|
| DOI / URN: |
10.1016/j.ymben.2018.01.001 |
|---|
| Katalog-ID: |
ELV041797566 |
|---|
| LEADER | 01000caa a22002652 4500 | ||
|---|---|---|---|
| 001 | ELV041797566 | ||
| 003 | DE-627 | ||
| 005 | 20230625235646.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 180726s2018 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1016/j.ymben.2018.01.001 |2 doi | |
| 028 | 5 | 2 | |a /cbs_pica/cbs_olc/import_discovery/elsevier/einzuspielen/GBV00000000001032.pica |
| 035 | |a (DE-627)ELV041797566 | ||
| 035 | |a (ELSEVIER)S1096-7176(17)30407-X | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 082 | 0 | 4 | |a 540 |q VZ |
| 082 | 0 | 4 | |a 610 |q VZ |
| 084 | |a 44.00 |2 bkl | ||
| 100 | 1 | |a Reifenrath, Mara |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Engineering of hydroxymandelate synthases and the aromatic amino acid pathway enables de novo biosynthesis of mandelic and 4-hydroxymandelic acid with Saccharomyces cerevisiae |
| 264 | 1 | |c 2018transfer abstract | |
| 300 | |a 9 | ||
| 336 | |a nicht spezifiziert |b zzz |2 rdacontent | ||
| 337 | |a nicht spezifiziert |b z |2 rdamedia | ||
| 338 | |a nicht spezifiziert |b zu |2 rdacarrier | ||
| 520 | |a Mandelic acid (MA) and 4-hydroxymandelic acid (HMA) are valuable specialty chemicals used as precursors for flavors as well as for cosmetic and pharmaceutical purposes. Today they are mainly synthesized chemically. Their synthesis through microbial fermentation would allow for environmentally sustainable production. In this work, we engineered the yeast Saccharomyces cerevisiae for high-level production of MA and HMA. Expressing the hydroxymandelate synthase from Amycolatopsis orientalis in a yeast wild type strain resulted in the production of 119mg/L HMA from glucose. As the enzyme also accepts phenylpyruvate as a substrate aside from its native substrate 4-hydroxyphenylpyruvate, 0.7mg/L MA was also produced. Preventing binding of 4-hydroxyphenylpyruvate to the hydroxymandelate synthase by introducing a S201V replacement in its substrate binding site nearly completely prevented HMA production but increased MA production only 3.5-fold. To further increase HMA and MA production, the aromatic amino acid pathway was engineered. We increased the precursor supply by introducing modifications in the shikimic acid pathway (ARO1↑, ARO3 K222L ↑, ARO4 K220L ↑) and reducing flux into the Ehrlich pathway (aro10Δ), and thereby enhanced the HMA titer to 465mg/L and the MA titer to 2.9mg/L. A further increase in HMA and MA titers was achieved by replacing the hydroxymandelate synthase from A. orientalis with the corresponding enzyme from Nocardia uniformis. Subsequently, we introduced additional deletions to block the competing tryptophan branch (trp2Δ), to further decrease flux into the Ehrlich pathway (pdc5Δ) and to avoid transamination of phenylpyruvate and 4-hydroxyphenylpyruvate (aro8Δ, aro9Δ). We achieved more than 1g/L 4-hydroxymandelate when additionally preventing formation of phenylpyruvate by deleting PHA2. When deleting TYR1 to prevent formation of 4-hydroxyphenylpyruvate instead, an MA titer of 236mg/L was achieved. This is a more than 200-fold increase in MA production compared to the wild type strain expressing the hydroxymandelate synthase from A. orientalis. Finally, we showed that S. cerevisiae tolerates HMA and MA to concentrations as high as 3g/L and 7.5g/L, respectively. Our results demonstrate that S. cerevisiae is a promising host for sustainable MA and HMA production. | ||
| 520 | |a Mandelic acid (MA) and 4-hydroxymandelic acid (HMA) are valuable specialty chemicals used as precursors for flavors as well as for cosmetic and pharmaceutical purposes. Today they are mainly synthesized chemically. Their synthesis through microbial fermentation would allow for environmentally sustainable production. In this work, we engineered the yeast Saccharomyces cerevisiae for high-level production of MA and HMA. Expressing the hydroxymandelate synthase from Amycolatopsis orientalis in a yeast wild type strain resulted in the production of 119mg/L HMA from glucose. As the enzyme also accepts phenylpyruvate as a substrate aside from its native substrate 4-hydroxyphenylpyruvate, 0.7mg/L MA was also produced. Preventing binding of 4-hydroxyphenylpyruvate to the hydroxymandelate synthase by introducing a S201V replacement in its substrate binding site nearly completely prevented HMA production but increased MA production only 3.5-fold. To further increase HMA and MA production, the aromatic amino acid pathway was engineered. We increased the precursor supply by introducing modifications in the shikimic acid pathway (ARO1↑, ARO3 K222L ↑, ARO4 K220L ↑) and reducing flux into the Ehrlich pathway (aro10Δ), and thereby enhanced the HMA titer to 465mg/L and the MA titer to 2.9mg/L. A further increase in HMA and MA titers was achieved by replacing the hydroxymandelate synthase from A. orientalis with the corresponding enzyme from Nocardia uniformis. Subsequently, we introduced additional deletions to block the competing tryptophan branch (trp2Δ), to further decrease flux into the Ehrlich pathway (pdc5Δ) and to avoid transamination of phenylpyruvate and 4-hydroxyphenylpyruvate (aro8Δ, aro9Δ). We achieved more than 1g/L 4-hydroxymandelate when additionally preventing formation of phenylpyruvate by deleting PHA2. When deleting TYR1 to prevent formation of 4-hydroxyphenylpyruvate instead, an MA titer of 236mg/L was achieved. This is a more than 200-fold increase in MA production compared to the wild type strain expressing the hydroxymandelate synthase from A. orientalis. Finally, we showed that S. cerevisiae tolerates HMA and MA to concentrations as high as 3g/L and 7.5g/L, respectively. Our results demonstrate that S. cerevisiae is a promising host for sustainable MA and HMA production. | ||
| 650 | 7 | |a PAA |2 Elsevier | |
| 650 | 7 | |a MA |2 Elsevier | |
| 650 | 7 | |a HmaS |2 Elsevier | |
| 650 | 7 | |a PPY |2 Elsevier | |
| 650 | 7 | |a HPP |2 Elsevier | |
| 650 | 7 | |a pPET |2 Elsevier | |
| 650 | 7 | |a HMA |2 Elsevier | |
| 650 | 7 | |a PET |2 Elsevier | |
| 700 | 1 | |a Boles, Eckhard |4 oth | |
| 773 | 0 | 8 | |i Enthalten in |n Academic Press |a Gao, Mengyao ELSEVIER |t Inhibition on polysulfides dissolve during the discharge-charge by using fish-scale-based porous carbon for lithium-sulfur battery |d 2014transfer abstract |g Orlando, Fla |w (DE-627)ELV012555568 |
| 773 | 1 | 8 | |g volume:45 |g year:2018 |g pages:246-254 |g extent:9 |
| 856 | 4 | 0 | |u https://doi.org/10.1016/j.ymben.2018.01.001 |3 Volltext |
| 912 | |a GBV_USEFLAG_U | ||
| 912 | |a GBV_ELV | ||
| 912 | |a SYSFLAG_U | ||
| 912 | |a SSG-OLC-PHA | ||
| 912 | |a GBV_ILN_20 | ||
| 912 | |a GBV_ILN_22 | ||
| 912 | |a GBV_ILN_40 | ||
| 912 | |a GBV_ILN_70 | ||
| 936 | b | k | |a 44.00 |j Medizin: Allgemeines |q VZ |
| 951 | |a AR | ||
| 952 | |d 45 |j 2018 |h 246-254 |g 9 | ||
| author_variant |
m r mr |
|---|---|
| matchkey_str |
reifenrathmaraboleseckhard:2018----:niernohdoyadltsnhssntermtcmnaiptwynbednvboyteiomneiadhdoy |
| hierarchy_sort_str |
2018transfer abstract |
| bklnumber |
44.00 |
| publishDate |
2018 |
| allfields |
10.1016/j.ymben.2018.01.001 doi /cbs_pica/cbs_olc/import_discovery/elsevier/einzuspielen/GBV00000000001032.pica (DE-627)ELV041797566 (ELSEVIER)S1096-7176(17)30407-X DE-627 ger DE-627 rakwb eng 540 VZ 610 VZ 44.00 bkl Reifenrath, Mara verfasserin aut Engineering of hydroxymandelate synthases and the aromatic amino acid pathway enables de novo biosynthesis of mandelic and 4-hydroxymandelic acid with Saccharomyces cerevisiae 2018transfer abstract 9 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier Mandelic acid (MA) and 4-hydroxymandelic acid (HMA) are valuable specialty chemicals used as precursors for flavors as well as for cosmetic and pharmaceutical purposes. Today they are mainly synthesized chemically. Their synthesis through microbial fermentation would allow for environmentally sustainable production. In this work, we engineered the yeast Saccharomyces cerevisiae for high-level production of MA and HMA. Expressing the hydroxymandelate synthase from Amycolatopsis orientalis in a yeast wild type strain resulted in the production of 119mg/L HMA from glucose. As the enzyme also accepts phenylpyruvate as a substrate aside from its native substrate 4-hydroxyphenylpyruvate, 0.7mg/L MA was also produced. Preventing binding of 4-hydroxyphenylpyruvate to the hydroxymandelate synthase by introducing a S201V replacement in its substrate binding site nearly completely prevented HMA production but increased MA production only 3.5-fold. To further increase HMA and MA production, the aromatic amino acid pathway was engineered. We increased the precursor supply by introducing modifications in the shikimic acid pathway (ARO1↑, ARO3 K222L ↑, ARO4 K220L ↑) and reducing flux into the Ehrlich pathway (aro10Δ), and thereby enhanced the HMA titer to 465mg/L and the MA titer to 2.9mg/L. A further increase in HMA and MA titers was achieved by replacing the hydroxymandelate synthase from A. orientalis with the corresponding enzyme from Nocardia uniformis. Subsequently, we introduced additional deletions to block the competing tryptophan branch (trp2Δ), to further decrease flux into the Ehrlich pathway (pdc5Δ) and to avoid transamination of phenylpyruvate and 4-hydroxyphenylpyruvate (aro8Δ, aro9Δ). We achieved more than 1g/L 4-hydroxymandelate when additionally preventing formation of phenylpyruvate by deleting PHA2. When deleting TYR1 to prevent formation of 4-hydroxyphenylpyruvate instead, an MA titer of 236mg/L was achieved. This is a more than 200-fold increase in MA production compared to the wild type strain expressing the hydroxymandelate synthase from A. orientalis. Finally, we showed that S. cerevisiae tolerates HMA and MA to concentrations as high as 3g/L and 7.5g/L, respectively. Our results demonstrate that S. cerevisiae is a promising host for sustainable MA and HMA production. Mandelic acid (MA) and 4-hydroxymandelic acid (HMA) are valuable specialty chemicals used as precursors for flavors as well as for cosmetic and pharmaceutical purposes. Today they are mainly synthesized chemically. Their synthesis through microbial fermentation would allow for environmentally sustainable production. In this work, we engineered the yeast Saccharomyces cerevisiae for high-level production of MA and HMA. Expressing the hydroxymandelate synthase from Amycolatopsis orientalis in a yeast wild type strain resulted in the production of 119mg/L HMA from glucose. As the enzyme also accepts phenylpyruvate as a substrate aside from its native substrate 4-hydroxyphenylpyruvate, 0.7mg/L MA was also produced. Preventing binding of 4-hydroxyphenylpyruvate to the hydroxymandelate synthase by introducing a S201V replacement in its substrate binding site nearly completely prevented HMA production but increased MA production only 3.5-fold. To further increase HMA and MA production, the aromatic amino acid pathway was engineered. We increased the precursor supply by introducing modifications in the shikimic acid pathway (ARO1↑, ARO3 K222L ↑, ARO4 K220L ↑) and reducing flux into the Ehrlich pathway (aro10Δ), and thereby enhanced the HMA titer to 465mg/L and the MA titer to 2.9mg/L. A further increase in HMA and MA titers was achieved by replacing the hydroxymandelate synthase from A. orientalis with the corresponding enzyme from Nocardia uniformis. Subsequently, we introduced additional deletions to block the competing tryptophan branch (trp2Δ), to further decrease flux into the Ehrlich pathway (pdc5Δ) and to avoid transamination of phenylpyruvate and 4-hydroxyphenylpyruvate (aro8Δ, aro9Δ). We achieved more than 1g/L 4-hydroxymandelate when additionally preventing formation of phenylpyruvate by deleting PHA2. When deleting TYR1 to prevent formation of 4-hydroxyphenylpyruvate instead, an MA titer of 236mg/L was achieved. This is a more than 200-fold increase in MA production compared to the wild type strain expressing the hydroxymandelate synthase from A. orientalis. Finally, we showed that S. cerevisiae tolerates HMA and MA to concentrations as high as 3g/L and 7.5g/L, respectively. Our results demonstrate that S. cerevisiae is a promising host for sustainable MA and HMA production. PAA Elsevier MA Elsevier HmaS Elsevier PPY Elsevier HPP Elsevier pPET Elsevier HMA Elsevier PET Elsevier Boles, Eckhard oth Enthalten in Academic Press Gao, Mengyao ELSEVIER Inhibition on polysulfides dissolve during the discharge-charge by using fish-scale-based porous carbon for lithium-sulfur battery 2014transfer abstract Orlando, Fla (DE-627)ELV012555568 volume:45 year:2018 pages:246-254 extent:9 https://doi.org/10.1016/j.ymben.2018.01.001 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_40 GBV_ILN_70 44.00 Medizin: Allgemeines VZ AR 45 2018 246-254 9 |
| spelling |
10.1016/j.ymben.2018.01.001 doi /cbs_pica/cbs_olc/import_discovery/elsevier/einzuspielen/GBV00000000001032.pica (DE-627)ELV041797566 (ELSEVIER)S1096-7176(17)30407-X DE-627 ger DE-627 rakwb eng 540 VZ 610 VZ 44.00 bkl Reifenrath, Mara verfasserin aut Engineering of hydroxymandelate synthases and the aromatic amino acid pathway enables de novo biosynthesis of mandelic and 4-hydroxymandelic acid with Saccharomyces cerevisiae 2018transfer abstract 9 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier Mandelic acid (MA) and 4-hydroxymandelic acid (HMA) are valuable specialty chemicals used as precursors for flavors as well as for cosmetic and pharmaceutical purposes. Today they are mainly synthesized chemically. Their synthesis through microbial fermentation would allow for environmentally sustainable production. In this work, we engineered the yeast Saccharomyces cerevisiae for high-level production of MA and HMA. Expressing the hydroxymandelate synthase from Amycolatopsis orientalis in a yeast wild type strain resulted in the production of 119mg/L HMA from glucose. As the enzyme also accepts phenylpyruvate as a substrate aside from its native substrate 4-hydroxyphenylpyruvate, 0.7mg/L MA was also produced. Preventing binding of 4-hydroxyphenylpyruvate to the hydroxymandelate synthase by introducing a S201V replacement in its substrate binding site nearly completely prevented HMA production but increased MA production only 3.5-fold. To further increase HMA and MA production, the aromatic amino acid pathway was engineered. We increased the precursor supply by introducing modifications in the shikimic acid pathway (ARO1↑, ARO3 K222L ↑, ARO4 K220L ↑) and reducing flux into the Ehrlich pathway (aro10Δ), and thereby enhanced the HMA titer to 465mg/L and the MA titer to 2.9mg/L. A further increase in HMA and MA titers was achieved by replacing the hydroxymandelate synthase from A. orientalis with the corresponding enzyme from Nocardia uniformis. Subsequently, we introduced additional deletions to block the competing tryptophan branch (trp2Δ), to further decrease flux into the Ehrlich pathway (pdc5Δ) and to avoid transamination of phenylpyruvate and 4-hydroxyphenylpyruvate (aro8Δ, aro9Δ). We achieved more than 1g/L 4-hydroxymandelate when additionally preventing formation of phenylpyruvate by deleting PHA2. When deleting TYR1 to prevent formation of 4-hydroxyphenylpyruvate instead, an MA titer of 236mg/L was achieved. This is a more than 200-fold increase in MA production compared to the wild type strain expressing the hydroxymandelate synthase from A. orientalis. Finally, we showed that S. cerevisiae tolerates HMA and MA to concentrations as high as 3g/L and 7.5g/L, respectively. Our results demonstrate that S. cerevisiae is a promising host for sustainable MA and HMA production. Mandelic acid (MA) and 4-hydroxymandelic acid (HMA) are valuable specialty chemicals used as precursors for flavors as well as for cosmetic and pharmaceutical purposes. Today they are mainly synthesized chemically. Their synthesis through microbial fermentation would allow for environmentally sustainable production. In this work, we engineered the yeast Saccharomyces cerevisiae for high-level production of MA and HMA. Expressing the hydroxymandelate synthase from Amycolatopsis orientalis in a yeast wild type strain resulted in the production of 119mg/L HMA from glucose. As the enzyme also accepts phenylpyruvate as a substrate aside from its native substrate 4-hydroxyphenylpyruvate, 0.7mg/L MA was also produced. Preventing binding of 4-hydroxyphenylpyruvate to the hydroxymandelate synthase by introducing a S201V replacement in its substrate binding site nearly completely prevented HMA production but increased MA production only 3.5-fold. To further increase HMA and MA production, the aromatic amino acid pathway was engineered. We increased the precursor supply by introducing modifications in the shikimic acid pathway (ARO1↑, ARO3 K222L ↑, ARO4 K220L ↑) and reducing flux into the Ehrlich pathway (aro10Δ), and thereby enhanced the HMA titer to 465mg/L and the MA titer to 2.9mg/L. A further increase in HMA and MA titers was achieved by replacing the hydroxymandelate synthase from A. orientalis with the corresponding enzyme from Nocardia uniformis. Subsequently, we introduced additional deletions to block the competing tryptophan branch (trp2Δ), to further decrease flux into the Ehrlich pathway (pdc5Δ) and to avoid transamination of phenylpyruvate and 4-hydroxyphenylpyruvate (aro8Δ, aro9Δ). We achieved more than 1g/L 4-hydroxymandelate when additionally preventing formation of phenylpyruvate by deleting PHA2. When deleting TYR1 to prevent formation of 4-hydroxyphenylpyruvate instead, an MA titer of 236mg/L was achieved. This is a more than 200-fold increase in MA production compared to the wild type strain expressing the hydroxymandelate synthase from A. orientalis. Finally, we showed that S. cerevisiae tolerates HMA and MA to concentrations as high as 3g/L and 7.5g/L, respectively. Our results demonstrate that S. cerevisiae is a promising host for sustainable MA and HMA production. PAA Elsevier MA Elsevier HmaS Elsevier PPY Elsevier HPP Elsevier pPET Elsevier HMA Elsevier PET Elsevier Boles, Eckhard oth Enthalten in Academic Press Gao, Mengyao ELSEVIER Inhibition on polysulfides dissolve during the discharge-charge by using fish-scale-based porous carbon for lithium-sulfur battery 2014transfer abstract Orlando, Fla (DE-627)ELV012555568 volume:45 year:2018 pages:246-254 extent:9 https://doi.org/10.1016/j.ymben.2018.01.001 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_40 GBV_ILN_70 44.00 Medizin: Allgemeines VZ AR 45 2018 246-254 9 |
| allfields_unstemmed |
10.1016/j.ymben.2018.01.001 doi /cbs_pica/cbs_olc/import_discovery/elsevier/einzuspielen/GBV00000000001032.pica (DE-627)ELV041797566 (ELSEVIER)S1096-7176(17)30407-X DE-627 ger DE-627 rakwb eng 540 VZ 610 VZ 44.00 bkl Reifenrath, Mara verfasserin aut Engineering of hydroxymandelate synthases and the aromatic amino acid pathway enables de novo biosynthesis of mandelic and 4-hydroxymandelic acid with Saccharomyces cerevisiae 2018transfer abstract 9 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier Mandelic acid (MA) and 4-hydroxymandelic acid (HMA) are valuable specialty chemicals used as precursors for flavors as well as for cosmetic and pharmaceutical purposes. Today they are mainly synthesized chemically. Their synthesis through microbial fermentation would allow for environmentally sustainable production. In this work, we engineered the yeast Saccharomyces cerevisiae for high-level production of MA and HMA. Expressing the hydroxymandelate synthase from Amycolatopsis orientalis in a yeast wild type strain resulted in the production of 119mg/L HMA from glucose. As the enzyme also accepts phenylpyruvate as a substrate aside from its native substrate 4-hydroxyphenylpyruvate, 0.7mg/L MA was also produced. Preventing binding of 4-hydroxyphenylpyruvate to the hydroxymandelate synthase by introducing a S201V replacement in its substrate binding site nearly completely prevented HMA production but increased MA production only 3.5-fold. To further increase HMA and MA production, the aromatic amino acid pathway was engineered. We increased the precursor supply by introducing modifications in the shikimic acid pathway (ARO1↑, ARO3 K222L ↑, ARO4 K220L ↑) and reducing flux into the Ehrlich pathway (aro10Δ), and thereby enhanced the HMA titer to 465mg/L and the MA titer to 2.9mg/L. A further increase in HMA and MA titers was achieved by replacing the hydroxymandelate synthase from A. orientalis with the corresponding enzyme from Nocardia uniformis. Subsequently, we introduced additional deletions to block the competing tryptophan branch (trp2Δ), to further decrease flux into the Ehrlich pathway (pdc5Δ) and to avoid transamination of phenylpyruvate and 4-hydroxyphenylpyruvate (aro8Δ, aro9Δ). We achieved more than 1g/L 4-hydroxymandelate when additionally preventing formation of phenylpyruvate by deleting PHA2. When deleting TYR1 to prevent formation of 4-hydroxyphenylpyruvate instead, an MA titer of 236mg/L was achieved. This is a more than 200-fold increase in MA production compared to the wild type strain expressing the hydroxymandelate synthase from A. orientalis. Finally, we showed that S. cerevisiae tolerates HMA and MA to concentrations as high as 3g/L and 7.5g/L, respectively. Our results demonstrate that S. cerevisiae is a promising host for sustainable MA and HMA production. Mandelic acid (MA) and 4-hydroxymandelic acid (HMA) are valuable specialty chemicals used as precursors for flavors as well as for cosmetic and pharmaceutical purposes. Today they are mainly synthesized chemically. Their synthesis through microbial fermentation would allow for environmentally sustainable production. In this work, we engineered the yeast Saccharomyces cerevisiae for high-level production of MA and HMA. Expressing the hydroxymandelate synthase from Amycolatopsis orientalis in a yeast wild type strain resulted in the production of 119mg/L HMA from glucose. As the enzyme also accepts phenylpyruvate as a substrate aside from its native substrate 4-hydroxyphenylpyruvate, 0.7mg/L MA was also produced. Preventing binding of 4-hydroxyphenylpyruvate to the hydroxymandelate synthase by introducing a S201V replacement in its substrate binding site nearly completely prevented HMA production but increased MA production only 3.5-fold. To further increase HMA and MA production, the aromatic amino acid pathway was engineered. We increased the precursor supply by introducing modifications in the shikimic acid pathway (ARO1↑, ARO3 K222L ↑, ARO4 K220L ↑) and reducing flux into the Ehrlich pathway (aro10Δ), and thereby enhanced the HMA titer to 465mg/L and the MA titer to 2.9mg/L. A further increase in HMA and MA titers was achieved by replacing the hydroxymandelate synthase from A. orientalis with the corresponding enzyme from Nocardia uniformis. Subsequently, we introduced additional deletions to block the competing tryptophan branch (trp2Δ), to further decrease flux into the Ehrlich pathway (pdc5Δ) and to avoid transamination of phenylpyruvate and 4-hydroxyphenylpyruvate (aro8Δ, aro9Δ). We achieved more than 1g/L 4-hydroxymandelate when additionally preventing formation of phenylpyruvate by deleting PHA2. When deleting TYR1 to prevent formation of 4-hydroxyphenylpyruvate instead, an MA titer of 236mg/L was achieved. This is a more than 200-fold increase in MA production compared to the wild type strain expressing the hydroxymandelate synthase from A. orientalis. Finally, we showed that S. cerevisiae tolerates HMA and MA to concentrations as high as 3g/L and 7.5g/L, respectively. Our results demonstrate that S. cerevisiae is a promising host for sustainable MA and HMA production. PAA Elsevier MA Elsevier HmaS Elsevier PPY Elsevier HPP Elsevier pPET Elsevier HMA Elsevier PET Elsevier Boles, Eckhard oth Enthalten in Academic Press Gao, Mengyao ELSEVIER Inhibition on polysulfides dissolve during the discharge-charge by using fish-scale-based porous carbon for lithium-sulfur battery 2014transfer abstract Orlando, Fla (DE-627)ELV012555568 volume:45 year:2018 pages:246-254 extent:9 https://doi.org/10.1016/j.ymben.2018.01.001 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_40 GBV_ILN_70 44.00 Medizin: Allgemeines VZ AR 45 2018 246-254 9 |
| allfieldsGer |
10.1016/j.ymben.2018.01.001 doi /cbs_pica/cbs_olc/import_discovery/elsevier/einzuspielen/GBV00000000001032.pica (DE-627)ELV041797566 (ELSEVIER)S1096-7176(17)30407-X DE-627 ger DE-627 rakwb eng 540 VZ 610 VZ 44.00 bkl Reifenrath, Mara verfasserin aut Engineering of hydroxymandelate synthases and the aromatic amino acid pathway enables de novo biosynthesis of mandelic and 4-hydroxymandelic acid with Saccharomyces cerevisiae 2018transfer abstract 9 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier Mandelic acid (MA) and 4-hydroxymandelic acid (HMA) are valuable specialty chemicals used as precursors for flavors as well as for cosmetic and pharmaceutical purposes. Today they are mainly synthesized chemically. Their synthesis through microbial fermentation would allow for environmentally sustainable production. In this work, we engineered the yeast Saccharomyces cerevisiae for high-level production of MA and HMA. Expressing the hydroxymandelate synthase from Amycolatopsis orientalis in a yeast wild type strain resulted in the production of 119mg/L HMA from glucose. As the enzyme also accepts phenylpyruvate as a substrate aside from its native substrate 4-hydroxyphenylpyruvate, 0.7mg/L MA was also produced. Preventing binding of 4-hydroxyphenylpyruvate to the hydroxymandelate synthase by introducing a S201V replacement in its substrate binding site nearly completely prevented HMA production but increased MA production only 3.5-fold. To further increase HMA and MA production, the aromatic amino acid pathway was engineered. We increased the precursor supply by introducing modifications in the shikimic acid pathway (ARO1↑, ARO3 K222L ↑, ARO4 K220L ↑) and reducing flux into the Ehrlich pathway (aro10Δ), and thereby enhanced the HMA titer to 465mg/L and the MA titer to 2.9mg/L. A further increase in HMA and MA titers was achieved by replacing the hydroxymandelate synthase from A. orientalis with the corresponding enzyme from Nocardia uniformis. Subsequently, we introduced additional deletions to block the competing tryptophan branch (trp2Δ), to further decrease flux into the Ehrlich pathway (pdc5Δ) and to avoid transamination of phenylpyruvate and 4-hydroxyphenylpyruvate (aro8Δ, aro9Δ). We achieved more than 1g/L 4-hydroxymandelate when additionally preventing formation of phenylpyruvate by deleting PHA2. When deleting TYR1 to prevent formation of 4-hydroxyphenylpyruvate instead, an MA titer of 236mg/L was achieved. This is a more than 200-fold increase in MA production compared to the wild type strain expressing the hydroxymandelate synthase from A. orientalis. Finally, we showed that S. cerevisiae tolerates HMA and MA to concentrations as high as 3g/L and 7.5g/L, respectively. Our results demonstrate that S. cerevisiae is a promising host for sustainable MA and HMA production. Mandelic acid (MA) and 4-hydroxymandelic acid (HMA) are valuable specialty chemicals used as precursors for flavors as well as for cosmetic and pharmaceutical purposes. Today they are mainly synthesized chemically. Their synthesis through microbial fermentation would allow for environmentally sustainable production. In this work, we engineered the yeast Saccharomyces cerevisiae for high-level production of MA and HMA. Expressing the hydroxymandelate synthase from Amycolatopsis orientalis in a yeast wild type strain resulted in the production of 119mg/L HMA from glucose. As the enzyme also accepts phenylpyruvate as a substrate aside from its native substrate 4-hydroxyphenylpyruvate, 0.7mg/L MA was also produced. Preventing binding of 4-hydroxyphenylpyruvate to the hydroxymandelate synthase by introducing a S201V replacement in its substrate binding site nearly completely prevented HMA production but increased MA production only 3.5-fold. To further increase HMA and MA production, the aromatic amino acid pathway was engineered. We increased the precursor supply by introducing modifications in the shikimic acid pathway (ARO1↑, ARO3 K222L ↑, ARO4 K220L ↑) and reducing flux into the Ehrlich pathway (aro10Δ), and thereby enhanced the HMA titer to 465mg/L and the MA titer to 2.9mg/L. A further increase in HMA and MA titers was achieved by replacing the hydroxymandelate synthase from A. orientalis with the corresponding enzyme from Nocardia uniformis. Subsequently, we introduced additional deletions to block the competing tryptophan branch (trp2Δ), to further decrease flux into the Ehrlich pathway (pdc5Δ) and to avoid transamination of phenylpyruvate and 4-hydroxyphenylpyruvate (aro8Δ, aro9Δ). We achieved more than 1g/L 4-hydroxymandelate when additionally preventing formation of phenylpyruvate by deleting PHA2. When deleting TYR1 to prevent formation of 4-hydroxyphenylpyruvate instead, an MA titer of 236mg/L was achieved. This is a more than 200-fold increase in MA production compared to the wild type strain expressing the hydroxymandelate synthase from A. orientalis. Finally, we showed that S. cerevisiae tolerates HMA and MA to concentrations as high as 3g/L and 7.5g/L, respectively. Our results demonstrate that S. cerevisiae is a promising host for sustainable MA and HMA production. PAA Elsevier MA Elsevier HmaS Elsevier PPY Elsevier HPP Elsevier pPET Elsevier HMA Elsevier PET Elsevier Boles, Eckhard oth Enthalten in Academic Press Gao, Mengyao ELSEVIER Inhibition on polysulfides dissolve during the discharge-charge by using fish-scale-based porous carbon for lithium-sulfur battery 2014transfer abstract Orlando, Fla (DE-627)ELV012555568 volume:45 year:2018 pages:246-254 extent:9 https://doi.org/10.1016/j.ymben.2018.01.001 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_40 GBV_ILN_70 44.00 Medizin: Allgemeines VZ AR 45 2018 246-254 9 |
| allfieldsSound |
10.1016/j.ymben.2018.01.001 doi /cbs_pica/cbs_olc/import_discovery/elsevier/einzuspielen/GBV00000000001032.pica (DE-627)ELV041797566 (ELSEVIER)S1096-7176(17)30407-X DE-627 ger DE-627 rakwb eng 540 VZ 610 VZ 44.00 bkl Reifenrath, Mara verfasserin aut Engineering of hydroxymandelate synthases and the aromatic amino acid pathway enables de novo biosynthesis of mandelic and 4-hydroxymandelic acid with Saccharomyces cerevisiae 2018transfer abstract 9 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier Mandelic acid (MA) and 4-hydroxymandelic acid (HMA) are valuable specialty chemicals used as precursors for flavors as well as for cosmetic and pharmaceutical purposes. Today they are mainly synthesized chemically. Their synthesis through microbial fermentation would allow for environmentally sustainable production. In this work, we engineered the yeast Saccharomyces cerevisiae for high-level production of MA and HMA. Expressing the hydroxymandelate synthase from Amycolatopsis orientalis in a yeast wild type strain resulted in the production of 119mg/L HMA from glucose. As the enzyme also accepts phenylpyruvate as a substrate aside from its native substrate 4-hydroxyphenylpyruvate, 0.7mg/L MA was also produced. Preventing binding of 4-hydroxyphenylpyruvate to the hydroxymandelate synthase by introducing a S201V replacement in its substrate binding site nearly completely prevented HMA production but increased MA production only 3.5-fold. To further increase HMA and MA production, the aromatic amino acid pathway was engineered. We increased the precursor supply by introducing modifications in the shikimic acid pathway (ARO1↑, ARO3 K222L ↑, ARO4 K220L ↑) and reducing flux into the Ehrlich pathway (aro10Δ), and thereby enhanced the HMA titer to 465mg/L and the MA titer to 2.9mg/L. A further increase in HMA and MA titers was achieved by replacing the hydroxymandelate synthase from A. orientalis with the corresponding enzyme from Nocardia uniformis. Subsequently, we introduced additional deletions to block the competing tryptophan branch (trp2Δ), to further decrease flux into the Ehrlich pathway (pdc5Δ) and to avoid transamination of phenylpyruvate and 4-hydroxyphenylpyruvate (aro8Δ, aro9Δ). We achieved more than 1g/L 4-hydroxymandelate when additionally preventing formation of phenylpyruvate by deleting PHA2. When deleting TYR1 to prevent formation of 4-hydroxyphenylpyruvate instead, an MA titer of 236mg/L was achieved. This is a more than 200-fold increase in MA production compared to the wild type strain expressing the hydroxymandelate synthase from A. orientalis. Finally, we showed that S. cerevisiae tolerates HMA and MA to concentrations as high as 3g/L and 7.5g/L, respectively. Our results demonstrate that S. cerevisiae is a promising host for sustainable MA and HMA production. Mandelic acid (MA) and 4-hydroxymandelic acid (HMA) are valuable specialty chemicals used as precursors for flavors as well as for cosmetic and pharmaceutical purposes. Today they are mainly synthesized chemically. Their synthesis through microbial fermentation would allow for environmentally sustainable production. In this work, we engineered the yeast Saccharomyces cerevisiae for high-level production of MA and HMA. Expressing the hydroxymandelate synthase from Amycolatopsis orientalis in a yeast wild type strain resulted in the production of 119mg/L HMA from glucose. As the enzyme also accepts phenylpyruvate as a substrate aside from its native substrate 4-hydroxyphenylpyruvate, 0.7mg/L MA was also produced. Preventing binding of 4-hydroxyphenylpyruvate to the hydroxymandelate synthase by introducing a S201V replacement in its substrate binding site nearly completely prevented HMA production but increased MA production only 3.5-fold. To further increase HMA and MA production, the aromatic amino acid pathway was engineered. We increased the precursor supply by introducing modifications in the shikimic acid pathway (ARO1↑, ARO3 K222L ↑, ARO4 K220L ↑) and reducing flux into the Ehrlich pathway (aro10Δ), and thereby enhanced the HMA titer to 465mg/L and the MA titer to 2.9mg/L. A further increase in HMA and MA titers was achieved by replacing the hydroxymandelate synthase from A. orientalis with the corresponding enzyme from Nocardia uniformis. Subsequently, we introduced additional deletions to block the competing tryptophan branch (trp2Δ), to further decrease flux into the Ehrlich pathway (pdc5Δ) and to avoid transamination of phenylpyruvate and 4-hydroxyphenylpyruvate (aro8Δ, aro9Δ). We achieved more than 1g/L 4-hydroxymandelate when additionally preventing formation of phenylpyruvate by deleting PHA2. When deleting TYR1 to prevent formation of 4-hydroxyphenylpyruvate instead, an MA titer of 236mg/L was achieved. This is a more than 200-fold increase in MA production compared to the wild type strain expressing the hydroxymandelate synthase from A. orientalis. Finally, we showed that S. cerevisiae tolerates HMA and MA to concentrations as high as 3g/L and 7.5g/L, respectively. Our results demonstrate that S. cerevisiae is a promising host for sustainable MA and HMA production. PAA Elsevier MA Elsevier HmaS Elsevier PPY Elsevier HPP Elsevier pPET Elsevier HMA Elsevier PET Elsevier Boles, Eckhard oth Enthalten in Academic Press Gao, Mengyao ELSEVIER Inhibition on polysulfides dissolve during the discharge-charge by using fish-scale-based porous carbon for lithium-sulfur battery 2014transfer abstract Orlando, Fla (DE-627)ELV012555568 volume:45 year:2018 pages:246-254 extent:9 https://doi.org/10.1016/j.ymben.2018.01.001 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_40 GBV_ILN_70 44.00 Medizin: Allgemeines VZ AR 45 2018 246-254 9 |
| language |
English |
| source |
Enthalten in Inhibition on polysulfides dissolve during the discharge-charge by using fish-scale-based porous carbon for lithium-sulfur battery Orlando, Fla volume:45 year:2018 pages:246-254 extent:9 |
| sourceStr |
Enthalten in Inhibition on polysulfides dissolve during the discharge-charge by using fish-scale-based porous carbon for lithium-sulfur battery Orlando, Fla volume:45 year:2018 pages:246-254 extent:9 |
| format_phy_str_mv |
Article |
| bklname |
Medizin: Allgemeines |
| institution |
findex.gbv.de |
| topic_facet |
PAA MA HmaS PPY HPP pPET HMA PET |
| dewey-raw |
540 |
| isfreeaccess_bool |
false |
| container_title |
Inhibition on polysulfides dissolve during the discharge-charge by using fish-scale-based porous carbon for lithium-sulfur battery |
| authorswithroles_txt_mv |
Reifenrath, Mara @@aut@@ Boles, Eckhard @@oth@@ |
| publishDateDaySort_date |
2018-01-01T00:00:00Z |
| hierarchy_top_id |
ELV012555568 |
| dewey-sort |
3540 |
| id |
ELV041797566 |
| language_de |
englisch |
| fullrecord |
<?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01000caa a22002652 4500</leader><controlfield tag="001">ELV041797566</controlfield><controlfield tag="003">DE-627</controlfield><controlfield tag="005">20230625235646.0</controlfield><controlfield tag="007">cr uuu---uuuuu</controlfield><controlfield tag="008">180726s2018 xx |||||o 00| ||eng c</controlfield><datafield tag="024" ind1="7" ind2=" "><subfield code="a">10.1016/j.ymben.2018.01.001</subfield><subfield code="2">doi</subfield></datafield><datafield tag="028" ind1="5" ind2="2"><subfield code="a">/cbs_pica/cbs_olc/import_discovery/elsevier/einzuspielen/GBV00000000001032.pica</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-627)ELV041797566</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(ELSEVIER)S1096-7176(17)30407-X</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-627</subfield><subfield code="b">ger</subfield><subfield code="c">DE-627</subfield><subfield code="e">rakwb</subfield></datafield><datafield tag="041" ind1=" " ind2=" "><subfield code="a">eng</subfield></datafield><datafield tag="082" ind1="0" ind2="4"><subfield code="a">540</subfield><subfield code="q">VZ</subfield></datafield><datafield tag="082" ind1="0" ind2="4"><subfield code="a">610</subfield><subfield code="q">VZ</subfield></datafield><datafield tag="084" ind1=" " ind2=" "><subfield code="a">44.00</subfield><subfield code="2">bkl</subfield></datafield><datafield tag="100" ind1="1" ind2=" "><subfield code="a">Reifenrath, Mara</subfield><subfield code="e">verfasserin</subfield><subfield code="4">aut</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Engineering of hydroxymandelate synthases and the aromatic amino acid pathway enables de novo biosynthesis of mandelic and 4-hydroxymandelic acid with Saccharomyces cerevisiae</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="c">2018transfer abstract</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="a">9</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="a">nicht spezifiziert</subfield><subfield code="b">zzz</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="a">nicht spezifiziert</subfield><subfield code="b">z</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="a">nicht spezifiziert</subfield><subfield code="b">zu</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="520" ind1=" " ind2=" "><subfield code="a">Mandelic acid (MA) and 4-hydroxymandelic acid (HMA) are valuable specialty chemicals used as precursors for flavors as well as for cosmetic and pharmaceutical purposes. Today they are mainly synthesized chemically. Their synthesis through microbial fermentation would allow for environmentally sustainable production. In this work, we engineered the yeast Saccharomyces cerevisiae for high-level production of MA and HMA. Expressing the hydroxymandelate synthase from Amycolatopsis orientalis in a yeast wild type strain resulted in the production of 119mg/L HMA from glucose. As the enzyme also accepts phenylpyruvate as a substrate aside from its native substrate 4-hydroxyphenylpyruvate, 0.7mg/L MA was also produced. Preventing binding of 4-hydroxyphenylpyruvate to the hydroxymandelate synthase by introducing a S201V replacement in its substrate binding site nearly completely prevented HMA production but increased MA production only 3.5-fold. To further increase HMA and MA production, the aromatic amino acid pathway was engineered. We increased the precursor supply by introducing modifications in the shikimic acid pathway (ARO1↑, ARO3 K222L ↑, ARO4 K220L ↑) and reducing flux into the Ehrlich pathway (aro10Δ), and thereby enhanced the HMA titer to 465mg/L and the MA titer to 2.9mg/L. A further increase in HMA and MA titers was achieved by replacing the hydroxymandelate synthase from A. orientalis with the corresponding enzyme from Nocardia uniformis. Subsequently, we introduced additional deletions to block the competing tryptophan branch (trp2Δ), to further decrease flux into the Ehrlich pathway (pdc5Δ) and to avoid transamination of phenylpyruvate and 4-hydroxyphenylpyruvate (aro8Δ, aro9Δ). We achieved more than 1g/L 4-hydroxymandelate when additionally preventing formation of phenylpyruvate by deleting PHA2. When deleting TYR1 to prevent formation of 4-hydroxyphenylpyruvate instead, an MA titer of 236mg/L was achieved. This is a more than 200-fold increase in MA production compared to the wild type strain expressing the hydroxymandelate synthase from A. orientalis. Finally, we showed that S. cerevisiae tolerates HMA and MA to concentrations as high as 3g/L and 7.5g/L, respectively. Our results demonstrate that S. cerevisiae is a promising host for sustainable MA and HMA production.</subfield></datafield><datafield tag="520" ind1=" " ind2=" "><subfield code="a">Mandelic acid (MA) and 4-hydroxymandelic acid (HMA) are valuable specialty chemicals used as precursors for flavors as well as for cosmetic and pharmaceutical purposes. Today they are mainly synthesized chemically. Their synthesis through microbial fermentation would allow for environmentally sustainable production. In this work, we engineered the yeast Saccharomyces cerevisiae for high-level production of MA and HMA. Expressing the hydroxymandelate synthase from Amycolatopsis orientalis in a yeast wild type strain resulted in the production of 119mg/L HMA from glucose. As the enzyme also accepts phenylpyruvate as a substrate aside from its native substrate 4-hydroxyphenylpyruvate, 0.7mg/L MA was also produced. Preventing binding of 4-hydroxyphenylpyruvate to the hydroxymandelate synthase by introducing a S201V replacement in its substrate binding site nearly completely prevented HMA production but increased MA production only 3.5-fold. To further increase HMA and MA production, the aromatic amino acid pathway was engineered. We increased the precursor supply by introducing modifications in the shikimic acid pathway (ARO1↑, ARO3 K222L ↑, ARO4 K220L ↑) and reducing flux into the Ehrlich pathway (aro10Δ), and thereby enhanced the HMA titer to 465mg/L and the MA titer to 2.9mg/L. A further increase in HMA and MA titers was achieved by replacing the hydroxymandelate synthase from A. orientalis with the corresponding enzyme from Nocardia uniformis. Subsequently, we introduced additional deletions to block the competing tryptophan branch (trp2Δ), to further decrease flux into the Ehrlich pathway (pdc5Δ) and to avoid transamination of phenylpyruvate and 4-hydroxyphenylpyruvate (aro8Δ, aro9Δ). We achieved more than 1g/L 4-hydroxymandelate when additionally preventing formation of phenylpyruvate by deleting PHA2. When deleting TYR1 to prevent formation of 4-hydroxyphenylpyruvate instead, an MA titer of 236mg/L was achieved. This is a more than 200-fold increase in MA production compared to the wild type strain expressing the hydroxymandelate synthase from A. orientalis. Finally, we showed that S. cerevisiae tolerates HMA and MA to concentrations as high as 3g/L and 7.5g/L, respectively. Our results demonstrate that S. cerevisiae is a promising host for sustainable MA and HMA production.</subfield></datafield><datafield tag="650" ind1=" " ind2="7"><subfield code="a">PAA</subfield><subfield code="2">Elsevier</subfield></datafield><datafield tag="650" ind1=" " ind2="7"><subfield code="a">MA</subfield><subfield code="2">Elsevier</subfield></datafield><datafield tag="650" ind1=" " ind2="7"><subfield code="a">HmaS</subfield><subfield code="2">Elsevier</subfield></datafield><datafield tag="650" ind1=" " ind2="7"><subfield code="a">PPY</subfield><subfield code="2">Elsevier</subfield></datafield><datafield tag="650" ind1=" " ind2="7"><subfield code="a">HPP</subfield><subfield code="2">Elsevier</subfield></datafield><datafield tag="650" ind1=" " ind2="7"><subfield code="a">pPET</subfield><subfield code="2">Elsevier</subfield></datafield><datafield tag="650" ind1=" " ind2="7"><subfield code="a">HMA</subfield><subfield code="2">Elsevier</subfield></datafield><datafield tag="650" ind1=" " ind2="7"><subfield code="a">PET</subfield><subfield code="2">Elsevier</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Boles, Eckhard</subfield><subfield code="4">oth</subfield></datafield><datafield tag="773" ind1="0" ind2="8"><subfield code="i">Enthalten in</subfield><subfield code="n">Academic Press</subfield><subfield code="a">Gao, Mengyao ELSEVIER</subfield><subfield code="t">Inhibition on polysulfides dissolve during the discharge-charge by using fish-scale-based porous carbon for lithium-sulfur battery</subfield><subfield code="d">2014transfer abstract</subfield><subfield code="g">Orlando, Fla</subfield><subfield code="w">(DE-627)ELV012555568</subfield></datafield><datafield tag="773" ind1="1" ind2="8"><subfield code="g">volume:45</subfield><subfield code="g">year:2018</subfield><subfield code="g">pages:246-254</subfield><subfield code="g">extent:9</subfield></datafield><datafield tag="856" ind1="4" ind2="0"><subfield code="u">https://doi.org/10.1016/j.ymben.2018.01.001</subfield><subfield code="3">Volltext</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_USEFLAG_U</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_ELV</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">SYSFLAG_U</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">SSG-OLC-PHA</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_ILN_20</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_ILN_22</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_ILN_40</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_ILN_70</subfield></datafield><datafield tag="936" ind1="b" ind2="k"><subfield code="a">44.00</subfield><subfield code="j">Medizin: Allgemeines</subfield><subfield code="q">VZ</subfield></datafield><datafield tag="951" ind1=" " ind2=" "><subfield code="a">AR</subfield></datafield><datafield tag="952" ind1=" " ind2=" "><subfield code="d">45</subfield><subfield code="j">2018</subfield><subfield code="h">246-254</subfield><subfield code="g">9</subfield></datafield></record></collection>
|
| author |
Reifenrath, Mara |
| spellingShingle |
Reifenrath, Mara ddc 540 ddc 610 bkl 44.00 Elsevier PAA Elsevier MA Elsevier HmaS Elsevier PPY Elsevier HPP Elsevier pPET Elsevier HMA Elsevier PET Engineering of hydroxymandelate synthases and the aromatic amino acid pathway enables de novo biosynthesis of mandelic and 4-hydroxymandelic acid with Saccharomyces cerevisiae |
| authorStr |
Reifenrath, Mara |
| ppnlink_with_tag_str_mv |
@@773@@(DE-627)ELV012555568 |
| format |
electronic Article |
| dewey-ones |
540 - Chemistry & allied sciences 610 - Medicine & health |
| delete_txt_mv |
keep |
| author_role |
aut |
| collection |
elsevier |
| remote_str |
true |
| illustrated |
Not Illustrated |
| topic_title |
540 VZ 610 VZ 44.00 bkl Engineering of hydroxymandelate synthases and the aromatic amino acid pathway enables de novo biosynthesis of mandelic and 4-hydroxymandelic acid with Saccharomyces cerevisiae PAA Elsevier MA Elsevier HmaS Elsevier PPY Elsevier HPP Elsevier pPET Elsevier HMA Elsevier PET Elsevier |
| topic |
ddc 540 ddc 610 bkl 44.00 Elsevier PAA Elsevier MA Elsevier HmaS Elsevier PPY Elsevier HPP Elsevier pPET Elsevier HMA Elsevier PET |
| topic_unstemmed |
ddc 540 ddc 610 bkl 44.00 Elsevier PAA Elsevier MA Elsevier HmaS Elsevier PPY Elsevier HPP Elsevier pPET Elsevier HMA Elsevier PET |
| topic_browse |
ddc 540 ddc 610 bkl 44.00 Elsevier PAA Elsevier MA Elsevier HmaS Elsevier PPY Elsevier HPP Elsevier pPET Elsevier HMA Elsevier PET |
| format_facet |
Elektronische Aufsätze Aufsätze Elektronische Ressource |
| format_main_str_mv |
Text Zeitschrift/Artikel |
| carriertype_str_mv |
zu |
| author2_variant |
e b eb |
| hierarchy_parent_title |
Inhibition on polysulfides dissolve during the discharge-charge by using fish-scale-based porous carbon for lithium-sulfur battery |
| hierarchy_parent_id |
ELV012555568 |
| dewey-tens |
540 - Chemistry 610 - Medicine & health |
| hierarchy_top_title |
Inhibition on polysulfides dissolve during the discharge-charge by using fish-scale-based porous carbon for lithium-sulfur battery |
| isfreeaccess_txt |
false |
| familylinks_str_mv |
(DE-627)ELV012555568 |
| title |
Engineering of hydroxymandelate synthases and the aromatic amino acid pathway enables de novo biosynthesis of mandelic and 4-hydroxymandelic acid with Saccharomyces cerevisiae |
| ctrlnum |
(DE-627)ELV041797566 (ELSEVIER)S1096-7176(17)30407-X |
| title_full |
Engineering of hydroxymandelate synthases and the aromatic amino acid pathway enables de novo biosynthesis of mandelic and 4-hydroxymandelic acid with Saccharomyces cerevisiae |
| author_sort |
Reifenrath, Mara |
| journal |
Inhibition on polysulfides dissolve during the discharge-charge by using fish-scale-based porous carbon for lithium-sulfur battery |
| journalStr |
Inhibition on polysulfides dissolve during the discharge-charge by using fish-scale-based porous carbon for lithium-sulfur battery |
| lang_code |
eng |
| isOA_bool |
false |
| dewey-hundreds |
500 - Science 600 - Technology |
| recordtype |
marc |
| publishDateSort |
2018 |
| contenttype_str_mv |
zzz |
| container_start_page |
246 |
| author_browse |
Reifenrath, Mara |
| container_volume |
45 |
| physical |
9 |
| class |
540 VZ 610 VZ 44.00 bkl |
| format_se |
Elektronische Aufsätze |
| author-letter |
Reifenrath, Mara |
| doi_str_mv |
10.1016/j.ymben.2018.01.001 |
| dewey-full |
540 610 |
| title_sort |
engineering of hydroxymandelate synthases and the aromatic amino acid pathway enables de novo biosynthesis of mandelic and 4-hydroxymandelic acid with saccharomyces cerevisiae |
| title_auth |
Engineering of hydroxymandelate synthases and the aromatic amino acid pathway enables de novo biosynthesis of mandelic and 4-hydroxymandelic acid with Saccharomyces cerevisiae |
| abstract |
Mandelic acid (MA) and 4-hydroxymandelic acid (HMA) are valuable specialty chemicals used as precursors for flavors as well as for cosmetic and pharmaceutical purposes. Today they are mainly synthesized chemically. Their synthesis through microbial fermentation would allow for environmentally sustainable production. In this work, we engineered the yeast Saccharomyces cerevisiae for high-level production of MA and HMA. Expressing the hydroxymandelate synthase from Amycolatopsis orientalis in a yeast wild type strain resulted in the production of 119mg/L HMA from glucose. As the enzyme also accepts phenylpyruvate as a substrate aside from its native substrate 4-hydroxyphenylpyruvate, 0.7mg/L MA was also produced. Preventing binding of 4-hydroxyphenylpyruvate to the hydroxymandelate synthase by introducing a S201V replacement in its substrate binding site nearly completely prevented HMA production but increased MA production only 3.5-fold. To further increase HMA and MA production, the aromatic amino acid pathway was engineered. We increased the precursor supply by introducing modifications in the shikimic acid pathway (ARO1↑, ARO3 K222L ↑, ARO4 K220L ↑) and reducing flux into the Ehrlich pathway (aro10Δ), and thereby enhanced the HMA titer to 465mg/L and the MA titer to 2.9mg/L. A further increase in HMA and MA titers was achieved by replacing the hydroxymandelate synthase from A. orientalis with the corresponding enzyme from Nocardia uniformis. Subsequently, we introduced additional deletions to block the competing tryptophan branch (trp2Δ), to further decrease flux into the Ehrlich pathway (pdc5Δ) and to avoid transamination of phenylpyruvate and 4-hydroxyphenylpyruvate (aro8Δ, aro9Δ). We achieved more than 1g/L 4-hydroxymandelate when additionally preventing formation of phenylpyruvate by deleting PHA2. When deleting TYR1 to prevent formation of 4-hydroxyphenylpyruvate instead, an MA titer of 236mg/L was achieved. This is a more than 200-fold increase in MA production compared to the wild type strain expressing the hydroxymandelate synthase from A. orientalis. Finally, we showed that S. cerevisiae tolerates HMA and MA to concentrations as high as 3g/L and 7.5g/L, respectively. Our results demonstrate that S. cerevisiae is a promising host for sustainable MA and HMA production. |
| abstractGer |
Mandelic acid (MA) and 4-hydroxymandelic acid (HMA) are valuable specialty chemicals used as precursors for flavors as well as for cosmetic and pharmaceutical purposes. Today they are mainly synthesized chemically. Their synthesis through microbial fermentation would allow for environmentally sustainable production. In this work, we engineered the yeast Saccharomyces cerevisiae for high-level production of MA and HMA. Expressing the hydroxymandelate synthase from Amycolatopsis orientalis in a yeast wild type strain resulted in the production of 119mg/L HMA from glucose. As the enzyme also accepts phenylpyruvate as a substrate aside from its native substrate 4-hydroxyphenylpyruvate, 0.7mg/L MA was also produced. Preventing binding of 4-hydroxyphenylpyruvate to the hydroxymandelate synthase by introducing a S201V replacement in its substrate binding site nearly completely prevented HMA production but increased MA production only 3.5-fold. To further increase HMA and MA production, the aromatic amino acid pathway was engineered. We increased the precursor supply by introducing modifications in the shikimic acid pathway (ARO1↑, ARO3 K222L ↑, ARO4 K220L ↑) and reducing flux into the Ehrlich pathway (aro10Δ), and thereby enhanced the HMA titer to 465mg/L and the MA titer to 2.9mg/L. A further increase in HMA and MA titers was achieved by replacing the hydroxymandelate synthase from A. orientalis with the corresponding enzyme from Nocardia uniformis. Subsequently, we introduced additional deletions to block the competing tryptophan branch (trp2Δ), to further decrease flux into the Ehrlich pathway (pdc5Δ) and to avoid transamination of phenylpyruvate and 4-hydroxyphenylpyruvate (aro8Δ, aro9Δ). We achieved more than 1g/L 4-hydroxymandelate when additionally preventing formation of phenylpyruvate by deleting PHA2. When deleting TYR1 to prevent formation of 4-hydroxyphenylpyruvate instead, an MA titer of 236mg/L was achieved. This is a more than 200-fold increase in MA production compared to the wild type strain expressing the hydroxymandelate synthase from A. orientalis. Finally, we showed that S. cerevisiae tolerates HMA and MA to concentrations as high as 3g/L and 7.5g/L, respectively. Our results demonstrate that S. cerevisiae is a promising host for sustainable MA and HMA production. |
| abstract_unstemmed |
Mandelic acid (MA) and 4-hydroxymandelic acid (HMA) are valuable specialty chemicals used as precursors for flavors as well as for cosmetic and pharmaceutical purposes. Today they are mainly synthesized chemically. Their synthesis through microbial fermentation would allow for environmentally sustainable production. In this work, we engineered the yeast Saccharomyces cerevisiae for high-level production of MA and HMA. Expressing the hydroxymandelate synthase from Amycolatopsis orientalis in a yeast wild type strain resulted in the production of 119mg/L HMA from glucose. As the enzyme also accepts phenylpyruvate as a substrate aside from its native substrate 4-hydroxyphenylpyruvate, 0.7mg/L MA was also produced. Preventing binding of 4-hydroxyphenylpyruvate to the hydroxymandelate synthase by introducing a S201V replacement in its substrate binding site nearly completely prevented HMA production but increased MA production only 3.5-fold. To further increase HMA and MA production, the aromatic amino acid pathway was engineered. We increased the precursor supply by introducing modifications in the shikimic acid pathway (ARO1↑, ARO3 K222L ↑, ARO4 K220L ↑) and reducing flux into the Ehrlich pathway (aro10Δ), and thereby enhanced the HMA titer to 465mg/L and the MA titer to 2.9mg/L. A further increase in HMA and MA titers was achieved by replacing the hydroxymandelate synthase from A. orientalis with the corresponding enzyme from Nocardia uniformis. Subsequently, we introduced additional deletions to block the competing tryptophan branch (trp2Δ), to further decrease flux into the Ehrlich pathway (pdc5Δ) and to avoid transamination of phenylpyruvate and 4-hydroxyphenylpyruvate (aro8Δ, aro9Δ). We achieved more than 1g/L 4-hydroxymandelate when additionally preventing formation of phenylpyruvate by deleting PHA2. When deleting TYR1 to prevent formation of 4-hydroxyphenylpyruvate instead, an MA titer of 236mg/L was achieved. This is a more than 200-fold increase in MA production compared to the wild type strain expressing the hydroxymandelate synthase from A. orientalis. Finally, we showed that S. cerevisiae tolerates HMA and MA to concentrations as high as 3g/L and 7.5g/L, respectively. Our results demonstrate that S. cerevisiae is a promising host for sustainable MA and HMA production. |
| collection_details |
GBV_USEFLAG_U GBV_ELV SYSFLAG_U SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_40 GBV_ILN_70 |
| title_short |
Engineering of hydroxymandelate synthases and the aromatic amino acid pathway enables de novo biosynthesis of mandelic and 4-hydroxymandelic acid with Saccharomyces cerevisiae |
| url |
https://doi.org/10.1016/j.ymben.2018.01.001 |
| remote_bool |
true |
| author2 |
Boles, Eckhard |
| author2Str |
Boles, Eckhard |
| ppnlink |
ELV012555568 |
| mediatype_str_mv |
z |
| isOA_txt |
false |
| hochschulschrift_bool |
false |
| author2_role |
oth |
| doi_str |
10.1016/j.ymben.2018.01.001 |
| up_date |
2024-07-06T21:05:58.058Z |
| _version_ |
1803865248822722560 |
| fullrecord_marcxml |
<?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01000caa a22002652 4500</leader><controlfield tag="001">ELV041797566</controlfield><controlfield tag="003">DE-627</controlfield><controlfield tag="005">20230625235646.0</controlfield><controlfield tag="007">cr uuu---uuuuu</controlfield><controlfield tag="008">180726s2018 xx |||||o 00| ||eng c</controlfield><datafield tag="024" ind1="7" ind2=" "><subfield code="a">10.1016/j.ymben.2018.01.001</subfield><subfield code="2">doi</subfield></datafield><datafield tag="028" ind1="5" ind2="2"><subfield code="a">/cbs_pica/cbs_olc/import_discovery/elsevier/einzuspielen/GBV00000000001032.pica</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-627)ELV041797566</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(ELSEVIER)S1096-7176(17)30407-X</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-627</subfield><subfield code="b">ger</subfield><subfield code="c">DE-627</subfield><subfield code="e">rakwb</subfield></datafield><datafield tag="041" ind1=" " ind2=" "><subfield code="a">eng</subfield></datafield><datafield tag="082" ind1="0" ind2="4"><subfield code="a">540</subfield><subfield code="q">VZ</subfield></datafield><datafield tag="082" ind1="0" ind2="4"><subfield code="a">610</subfield><subfield code="q">VZ</subfield></datafield><datafield tag="084" ind1=" " ind2=" "><subfield code="a">44.00</subfield><subfield code="2">bkl</subfield></datafield><datafield tag="100" ind1="1" ind2=" "><subfield code="a">Reifenrath, Mara</subfield><subfield code="e">verfasserin</subfield><subfield code="4">aut</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Engineering of hydroxymandelate synthases and the aromatic amino acid pathway enables de novo biosynthesis of mandelic and 4-hydroxymandelic acid with Saccharomyces cerevisiae</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="c">2018transfer abstract</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="a">9</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="a">nicht spezifiziert</subfield><subfield code="b">zzz</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="a">nicht spezifiziert</subfield><subfield code="b">z</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="a">nicht spezifiziert</subfield><subfield code="b">zu</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="520" ind1=" " ind2=" "><subfield code="a">Mandelic acid (MA) and 4-hydroxymandelic acid (HMA) are valuable specialty chemicals used as precursors for flavors as well as for cosmetic and pharmaceutical purposes. Today they are mainly synthesized chemically. Their synthesis through microbial fermentation would allow for environmentally sustainable production. In this work, we engineered the yeast Saccharomyces cerevisiae for high-level production of MA and HMA. Expressing the hydroxymandelate synthase from Amycolatopsis orientalis in a yeast wild type strain resulted in the production of 119mg/L HMA from glucose. As the enzyme also accepts phenylpyruvate as a substrate aside from its native substrate 4-hydroxyphenylpyruvate, 0.7mg/L MA was also produced. Preventing binding of 4-hydroxyphenylpyruvate to the hydroxymandelate synthase by introducing a S201V replacement in its substrate binding site nearly completely prevented HMA production but increased MA production only 3.5-fold. To further increase HMA and MA production, the aromatic amino acid pathway was engineered. We increased the precursor supply by introducing modifications in the shikimic acid pathway (ARO1↑, ARO3 K222L ↑, ARO4 K220L ↑) and reducing flux into the Ehrlich pathway (aro10Δ), and thereby enhanced the HMA titer to 465mg/L and the MA titer to 2.9mg/L. A further increase in HMA and MA titers was achieved by replacing the hydroxymandelate synthase from A. orientalis with the corresponding enzyme from Nocardia uniformis. Subsequently, we introduced additional deletions to block the competing tryptophan branch (trp2Δ), to further decrease flux into the Ehrlich pathway (pdc5Δ) and to avoid transamination of phenylpyruvate and 4-hydroxyphenylpyruvate (aro8Δ, aro9Δ). We achieved more than 1g/L 4-hydroxymandelate when additionally preventing formation of phenylpyruvate by deleting PHA2. When deleting TYR1 to prevent formation of 4-hydroxyphenylpyruvate instead, an MA titer of 236mg/L was achieved. This is a more than 200-fold increase in MA production compared to the wild type strain expressing the hydroxymandelate synthase from A. orientalis. Finally, we showed that S. cerevisiae tolerates HMA and MA to concentrations as high as 3g/L and 7.5g/L, respectively. Our results demonstrate that S. cerevisiae is a promising host for sustainable MA and HMA production.</subfield></datafield><datafield tag="520" ind1=" " ind2=" "><subfield code="a">Mandelic acid (MA) and 4-hydroxymandelic acid (HMA) are valuable specialty chemicals used as precursors for flavors as well as for cosmetic and pharmaceutical purposes. Today they are mainly synthesized chemically. Their synthesis through microbial fermentation would allow for environmentally sustainable production. In this work, we engineered the yeast Saccharomyces cerevisiae for high-level production of MA and HMA. Expressing the hydroxymandelate synthase from Amycolatopsis orientalis in a yeast wild type strain resulted in the production of 119mg/L HMA from glucose. As the enzyme also accepts phenylpyruvate as a substrate aside from its native substrate 4-hydroxyphenylpyruvate, 0.7mg/L MA was also produced. Preventing binding of 4-hydroxyphenylpyruvate to the hydroxymandelate synthase by introducing a S201V replacement in its substrate binding site nearly completely prevented HMA production but increased MA production only 3.5-fold. To further increase HMA and MA production, the aromatic amino acid pathway was engineered. We increased the precursor supply by introducing modifications in the shikimic acid pathway (ARO1↑, ARO3 K222L ↑, ARO4 K220L ↑) and reducing flux into the Ehrlich pathway (aro10Δ), and thereby enhanced the HMA titer to 465mg/L and the MA titer to 2.9mg/L. A further increase in HMA and MA titers was achieved by replacing the hydroxymandelate synthase from A. orientalis with the corresponding enzyme from Nocardia uniformis. Subsequently, we introduced additional deletions to block the competing tryptophan branch (trp2Δ), to further decrease flux into the Ehrlich pathway (pdc5Δ) and to avoid transamination of phenylpyruvate and 4-hydroxyphenylpyruvate (aro8Δ, aro9Δ). We achieved more than 1g/L 4-hydroxymandelate when additionally preventing formation of phenylpyruvate by deleting PHA2. When deleting TYR1 to prevent formation of 4-hydroxyphenylpyruvate instead, an MA titer of 236mg/L was achieved. This is a more than 200-fold increase in MA production compared to the wild type strain expressing the hydroxymandelate synthase from A. orientalis. Finally, we showed that S. cerevisiae tolerates HMA and MA to concentrations as high as 3g/L and 7.5g/L, respectively. Our results demonstrate that S. cerevisiae is a promising host for sustainable MA and HMA production.</subfield></datafield><datafield tag="650" ind1=" " ind2="7"><subfield code="a">PAA</subfield><subfield code="2">Elsevier</subfield></datafield><datafield tag="650" ind1=" " ind2="7"><subfield code="a">MA</subfield><subfield code="2">Elsevier</subfield></datafield><datafield tag="650" ind1=" " ind2="7"><subfield code="a">HmaS</subfield><subfield code="2">Elsevier</subfield></datafield><datafield tag="650" ind1=" " ind2="7"><subfield code="a">PPY</subfield><subfield code="2">Elsevier</subfield></datafield><datafield tag="650" ind1=" " ind2="7"><subfield code="a">HPP</subfield><subfield code="2">Elsevier</subfield></datafield><datafield tag="650" ind1=" " ind2="7"><subfield code="a">pPET</subfield><subfield code="2">Elsevier</subfield></datafield><datafield tag="650" ind1=" " ind2="7"><subfield code="a">HMA</subfield><subfield code="2">Elsevier</subfield></datafield><datafield tag="650" ind1=" " ind2="7"><subfield code="a">PET</subfield><subfield code="2">Elsevier</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Boles, Eckhard</subfield><subfield code="4">oth</subfield></datafield><datafield tag="773" ind1="0" ind2="8"><subfield code="i">Enthalten in</subfield><subfield code="n">Academic Press</subfield><subfield code="a">Gao, Mengyao ELSEVIER</subfield><subfield code="t">Inhibition on polysulfides dissolve during the discharge-charge by using fish-scale-based porous carbon for lithium-sulfur battery</subfield><subfield code="d">2014transfer abstract</subfield><subfield code="g">Orlando, Fla</subfield><subfield code="w">(DE-627)ELV012555568</subfield></datafield><datafield tag="773" ind1="1" ind2="8"><subfield code="g">volume:45</subfield><subfield code="g">year:2018</subfield><subfield code="g">pages:246-254</subfield><subfield code="g">extent:9</subfield></datafield><datafield tag="856" ind1="4" ind2="0"><subfield code="u">https://doi.org/10.1016/j.ymben.2018.01.001</subfield><subfield code="3">Volltext</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_USEFLAG_U</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_ELV</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">SYSFLAG_U</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">SSG-OLC-PHA</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_ILN_20</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_ILN_22</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_ILN_40</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_ILN_70</subfield></datafield><datafield tag="936" ind1="b" ind2="k"><subfield code="a">44.00</subfield><subfield code="j">Medizin: Allgemeines</subfield><subfield code="q">VZ</subfield></datafield><datafield tag="951" ind1=" " ind2=" "><subfield code="a">AR</subfield></datafield><datafield tag="952" ind1=" " ind2=" "><subfield code="d">45</subfield><subfield code="j">2018</subfield><subfield code="h">246-254</subfield><subfield code="g">9</subfield></datafield></record></collection>
|
| score |
7.3993816 |