Enhancing thermostability and the structural characterization of Microbacterium saccharophilum K-1 β-fructofuranosidase
Abstract A β-fructofuranosidase from Microbacterium saccharophilum K-1 (formerly known as Arthrobacter sp. K-1) is useful for producing the sweetener lactosucrose ($ 4^{G} $-β-d-galactosylsucrose). Thermostability of the β-fructofuranosidase was enhanced by random mutagenesis and saturation mutagene...
Ausführliche Beschreibung
Gespeichert in:
| Autor*in: |
Ohta, Yukari [verfasserIn] |
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| Format: |
Artikel |
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| Sprache: |
Englisch |
| Erschienen: |
2014 |
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| Schlagwörter: |
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| Anmerkung: |
© Springer-Verlag Berlin Heidelberg 2014 |
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| Übergeordnetes Werk: |
Enthalten in: Applied microbiology and biotechnology - Springer Berlin Heidelberg, 1984, 98(2014), 15 vom: 15. März, Seite 6667-6677 |
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| Übergeordnetes Werk: |
volume:98 ; year:2014 ; number:15 ; day:15 ; month:03 ; pages:6667-6677 |
| Links: |
|---|
| DOI / URN: |
10.1007/s00253-014-5645-3 |
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OLC2050761058 |
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| 245 | 1 | 0 | |a Enhancing thermostability and the structural characterization of Microbacterium saccharophilum K-1 β-fructofuranosidase |
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| 520 | |a Abstract A β-fructofuranosidase from Microbacterium saccharophilum K-1 (formerly known as Arthrobacter sp. K-1) is useful for producing the sweetener lactosucrose ($ 4^{G} $-β-d-galactosylsucrose). Thermostability of the β-fructofuranosidase was enhanced by random mutagenesis and saturation mutagenesis. Clones with enhanced thermostability included mutations at residues Thr47, Ser200, Phe447, Phe470, and Pro500. In the highest stability mutant, T47S/S200T/F447P/F470Y/P500S, the half-life at 60 °C was 182 min, 16.5-fold longer than the wild-type enzyme. A comparison of the crystal structures of the full-length wild-type enzyme and three mutants showed that various mechanisms appear to be involved in thermostability enhancement. In particular, the replacement of Phe447 with Val or Pro induced a conformational change in an adjacent residue His477, which results in the formation of a new hydrogen bond in the enzyme. Although the thermostabilization mechanisms of the five residue mutations were explicable on the basis of the crystal structures, it appears to be difficult to predict which amino acid residues should be modified to obtain thermostabilized enzymes. | ||
| 650 | 4 | |a Thermostability | |
| 650 | 4 | |a Random mutagenesis | |
| 650 | 4 | |a β-Fructofuranosidase | |
| 650 | 4 | |a Lactosucrose | |
| 650 | 4 | |a GH68 | |
| 700 | 1 | |a Hatada, Yuji |4 aut | |
| 700 | 1 | |a Hidaka, Yuko |4 aut | |
| 700 | 1 | |a Shimane, Yasuhiro |4 aut | |
| 700 | 1 | |a Usui, Keiko |4 aut | |
| 700 | 1 | |a Ito, Tetsuya |4 aut | |
| 700 | 1 | |a Fujita, Koki |4 aut | |
| 700 | 1 | |a Yokoi, Gaku |4 aut | |
| 700 | 1 | |a Mori, Marina |4 aut | |
| 700 | 1 | |a Sato, Shona |4 aut | |
| 700 | 1 | |a Miyazaki, Takatsugu |4 aut | |
| 700 | 1 | |a Nishikawa, Atsushi |4 aut | |
| 700 | 1 | |a Tonozuka, Takashi |4 aut | |
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10.1007/s00253-014-5645-3 doi (DE-627)OLC2050761058 (DE-He213)s00253-014-5645-3-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Ohta, Yukari verfasserin aut Enhancing thermostability and the structural characterization of Microbacterium saccharophilum K-1 β-fructofuranosidase 2014 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag Berlin Heidelberg 2014 Abstract A β-fructofuranosidase from Microbacterium saccharophilum K-1 (formerly known as Arthrobacter sp. K-1) is useful for producing the sweetener lactosucrose ($ 4^{G} $-β-d-galactosylsucrose). Thermostability of the β-fructofuranosidase was enhanced by random mutagenesis and saturation mutagenesis. Clones with enhanced thermostability included mutations at residues Thr47, Ser200, Phe447, Phe470, and Pro500. In the highest stability mutant, T47S/S200T/F447P/F470Y/P500S, the half-life at 60 °C was 182 min, 16.5-fold longer than the wild-type enzyme. A comparison of the crystal structures of the full-length wild-type enzyme and three mutants showed that various mechanisms appear to be involved in thermostability enhancement. In particular, the replacement of Phe447 with Val or Pro induced a conformational change in an adjacent residue His477, which results in the formation of a new hydrogen bond in the enzyme. Although the thermostabilization mechanisms of the five residue mutations were explicable on the basis of the crystal structures, it appears to be difficult to predict which amino acid residues should be modified to obtain thermostabilized enzymes. Thermostability Random mutagenesis β-Fructofuranosidase Lactosucrose GH68 Hatada, Yuji aut Hidaka, Yuko aut Shimane, Yasuhiro aut Usui, Keiko aut Ito, Tetsuya aut Fujita, Koki aut Yokoi, Gaku aut Mori, Marina aut Sato, Shona aut Miyazaki, Takatsugu aut Nishikawa, Atsushi aut Tonozuka, Takashi aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 98(2014), 15 vom: 15. März, Seite 6667-6677 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:98 year:2014 number:15 day:15 month:03 pages:6667-6677 https://doi.org/10.1007/s00253-014-5645-3 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_70 GBV_ILN_130 GBV_ILN_267 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4082 GBV_ILN_4277 GBV_ILN_4305 AR 98 2014 15 15 03 6667-6677 |
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10.1007/s00253-014-5645-3 doi (DE-627)OLC2050761058 (DE-He213)s00253-014-5645-3-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Ohta, Yukari verfasserin aut Enhancing thermostability and the structural characterization of Microbacterium saccharophilum K-1 β-fructofuranosidase 2014 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag Berlin Heidelberg 2014 Abstract A β-fructofuranosidase from Microbacterium saccharophilum K-1 (formerly known as Arthrobacter sp. K-1) is useful for producing the sweetener lactosucrose ($ 4^{G} $-β-d-galactosylsucrose). Thermostability of the β-fructofuranosidase was enhanced by random mutagenesis and saturation mutagenesis. Clones with enhanced thermostability included mutations at residues Thr47, Ser200, Phe447, Phe470, and Pro500. In the highest stability mutant, T47S/S200T/F447P/F470Y/P500S, the half-life at 60 °C was 182 min, 16.5-fold longer than the wild-type enzyme. A comparison of the crystal structures of the full-length wild-type enzyme and three mutants showed that various mechanisms appear to be involved in thermostability enhancement. In particular, the replacement of Phe447 with Val or Pro induced a conformational change in an adjacent residue His477, which results in the formation of a new hydrogen bond in the enzyme. Although the thermostabilization mechanisms of the five residue mutations were explicable on the basis of the crystal structures, it appears to be difficult to predict which amino acid residues should be modified to obtain thermostabilized enzymes. Thermostability Random mutagenesis β-Fructofuranosidase Lactosucrose GH68 Hatada, Yuji aut Hidaka, Yuko aut Shimane, Yasuhiro aut Usui, Keiko aut Ito, Tetsuya aut Fujita, Koki aut Yokoi, Gaku aut Mori, Marina aut Sato, Shona aut Miyazaki, Takatsugu aut Nishikawa, Atsushi aut Tonozuka, Takashi aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 98(2014), 15 vom: 15. März, Seite 6667-6677 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:98 year:2014 number:15 day:15 month:03 pages:6667-6677 https://doi.org/10.1007/s00253-014-5645-3 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_70 GBV_ILN_130 GBV_ILN_267 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4082 GBV_ILN_4277 GBV_ILN_4305 AR 98 2014 15 15 03 6667-6677 |
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10.1007/s00253-014-5645-3 doi (DE-627)OLC2050761058 (DE-He213)s00253-014-5645-3-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Ohta, Yukari verfasserin aut Enhancing thermostability and the structural characterization of Microbacterium saccharophilum K-1 β-fructofuranosidase 2014 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag Berlin Heidelberg 2014 Abstract A β-fructofuranosidase from Microbacterium saccharophilum K-1 (formerly known as Arthrobacter sp. K-1) is useful for producing the sweetener lactosucrose ($ 4^{G} $-β-d-galactosylsucrose). Thermostability of the β-fructofuranosidase was enhanced by random mutagenesis and saturation mutagenesis. Clones with enhanced thermostability included mutations at residues Thr47, Ser200, Phe447, Phe470, and Pro500. In the highest stability mutant, T47S/S200T/F447P/F470Y/P500S, the half-life at 60 °C was 182 min, 16.5-fold longer than the wild-type enzyme. A comparison of the crystal structures of the full-length wild-type enzyme and three mutants showed that various mechanisms appear to be involved in thermostability enhancement. In particular, the replacement of Phe447 with Val or Pro induced a conformational change in an adjacent residue His477, which results in the formation of a new hydrogen bond in the enzyme. Although the thermostabilization mechanisms of the five residue mutations were explicable on the basis of the crystal structures, it appears to be difficult to predict which amino acid residues should be modified to obtain thermostabilized enzymes. Thermostability Random mutagenesis β-Fructofuranosidase Lactosucrose GH68 Hatada, Yuji aut Hidaka, Yuko aut Shimane, Yasuhiro aut Usui, Keiko aut Ito, Tetsuya aut Fujita, Koki aut Yokoi, Gaku aut Mori, Marina aut Sato, Shona aut Miyazaki, Takatsugu aut Nishikawa, Atsushi aut Tonozuka, Takashi aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 98(2014), 15 vom: 15. März, Seite 6667-6677 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:98 year:2014 number:15 day:15 month:03 pages:6667-6677 https://doi.org/10.1007/s00253-014-5645-3 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_70 GBV_ILN_130 GBV_ILN_267 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4082 GBV_ILN_4277 GBV_ILN_4305 AR 98 2014 15 15 03 6667-6677 |
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10.1007/s00253-014-5645-3 doi (DE-627)OLC2050761058 (DE-He213)s00253-014-5645-3-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Ohta, Yukari verfasserin aut Enhancing thermostability and the structural characterization of Microbacterium saccharophilum K-1 β-fructofuranosidase 2014 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag Berlin Heidelberg 2014 Abstract A β-fructofuranosidase from Microbacterium saccharophilum K-1 (formerly known as Arthrobacter sp. K-1) is useful for producing the sweetener lactosucrose ($ 4^{G} $-β-d-galactosylsucrose). Thermostability of the β-fructofuranosidase was enhanced by random mutagenesis and saturation mutagenesis. Clones with enhanced thermostability included mutations at residues Thr47, Ser200, Phe447, Phe470, and Pro500. In the highest stability mutant, T47S/S200T/F447P/F470Y/P500S, the half-life at 60 °C was 182 min, 16.5-fold longer than the wild-type enzyme. A comparison of the crystal structures of the full-length wild-type enzyme and three mutants showed that various mechanisms appear to be involved in thermostability enhancement. In particular, the replacement of Phe447 with Val or Pro induced a conformational change in an adjacent residue His477, which results in the formation of a new hydrogen bond in the enzyme. Although the thermostabilization mechanisms of the five residue mutations were explicable on the basis of the crystal structures, it appears to be difficult to predict which amino acid residues should be modified to obtain thermostabilized enzymes. Thermostability Random mutagenesis β-Fructofuranosidase Lactosucrose GH68 Hatada, Yuji aut Hidaka, Yuko aut Shimane, Yasuhiro aut Usui, Keiko aut Ito, Tetsuya aut Fujita, Koki aut Yokoi, Gaku aut Mori, Marina aut Sato, Shona aut Miyazaki, Takatsugu aut Nishikawa, Atsushi aut Tonozuka, Takashi aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 98(2014), 15 vom: 15. März, Seite 6667-6677 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:98 year:2014 number:15 day:15 month:03 pages:6667-6677 https://doi.org/10.1007/s00253-014-5645-3 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_70 GBV_ILN_130 GBV_ILN_267 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4082 GBV_ILN_4277 GBV_ILN_4305 AR 98 2014 15 15 03 6667-6677 |
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10.1007/s00253-014-5645-3 doi (DE-627)OLC2050761058 (DE-He213)s00253-014-5645-3-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Ohta, Yukari verfasserin aut Enhancing thermostability and the structural characterization of Microbacterium saccharophilum K-1 β-fructofuranosidase 2014 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag Berlin Heidelberg 2014 Abstract A β-fructofuranosidase from Microbacterium saccharophilum K-1 (formerly known as Arthrobacter sp. K-1) is useful for producing the sweetener lactosucrose ($ 4^{G} $-β-d-galactosylsucrose). Thermostability of the β-fructofuranosidase was enhanced by random mutagenesis and saturation mutagenesis. Clones with enhanced thermostability included mutations at residues Thr47, Ser200, Phe447, Phe470, and Pro500. In the highest stability mutant, T47S/S200T/F447P/F470Y/P500S, the half-life at 60 °C was 182 min, 16.5-fold longer than the wild-type enzyme. A comparison of the crystal structures of the full-length wild-type enzyme and three mutants showed that various mechanisms appear to be involved in thermostability enhancement. In particular, the replacement of Phe447 with Val or Pro induced a conformational change in an adjacent residue His477, which results in the formation of a new hydrogen bond in the enzyme. Although the thermostabilization mechanisms of the five residue mutations were explicable on the basis of the crystal structures, it appears to be difficult to predict which amino acid residues should be modified to obtain thermostabilized enzymes. Thermostability Random mutagenesis β-Fructofuranosidase Lactosucrose GH68 Hatada, Yuji aut Hidaka, Yuko aut Shimane, Yasuhiro aut Usui, Keiko aut Ito, Tetsuya aut Fujita, Koki aut Yokoi, Gaku aut Mori, Marina aut Sato, Shona aut Miyazaki, Takatsugu aut Nishikawa, Atsushi aut Tonozuka, Takashi aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 98(2014), 15 vom: 15. März, Seite 6667-6677 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:98 year:2014 number:15 day:15 month:03 pages:6667-6677 https://doi.org/10.1007/s00253-014-5645-3 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_70 GBV_ILN_130 GBV_ILN_267 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4082 GBV_ILN_4277 GBV_ILN_4305 AR 98 2014 15 15 03 6667-6677 |
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Enthalten in Applied microbiology and biotechnology 98(2014), 15 vom: 15. März, Seite 6667-6677 volume:98 year:2014 number:15 day:15 month:03 pages:6667-6677 |
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Enthalten in Applied microbiology and biotechnology 98(2014), 15 vom: 15. März, Seite 6667-6677 volume:98 year:2014 number:15 day:15 month:03 pages:6667-6677 |
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Ohta, Yukari @@aut@@ Hatada, Yuji @@aut@@ Hidaka, Yuko @@aut@@ Shimane, Yasuhiro @@aut@@ Usui, Keiko @@aut@@ Ito, Tetsuya @@aut@@ Fujita, Koki @@aut@@ Yokoi, Gaku @@aut@@ Mori, Marina @@aut@@ Sato, Shona @@aut@@ Miyazaki, Takatsugu @@aut@@ Nishikawa, Atsushi @@aut@@ Tonozuka, Takashi @@aut@@ |
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Ohta, Yukari Hatada, Yuji Hidaka, Yuko Shimane, Yasuhiro Usui, Keiko Ito, Tetsuya Fujita, Koki Yokoi, Gaku Mori, Marina Sato, Shona Miyazaki, Takatsugu Nishikawa, Atsushi Tonozuka, Takashi |
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enhancing thermostability and the structural characterization of microbacterium saccharophilum k-1 β-fructofuranosidase |
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Enhancing thermostability and the structural characterization of Microbacterium saccharophilum K-1 β-fructofuranosidase |
| abstract |
Abstract A β-fructofuranosidase from Microbacterium saccharophilum K-1 (formerly known as Arthrobacter sp. K-1) is useful for producing the sweetener lactosucrose ($ 4^{G} $-β-d-galactosylsucrose). Thermostability of the β-fructofuranosidase was enhanced by random mutagenesis and saturation mutagenesis. Clones with enhanced thermostability included mutations at residues Thr47, Ser200, Phe447, Phe470, and Pro500. In the highest stability mutant, T47S/S200T/F447P/F470Y/P500S, the half-life at 60 °C was 182 min, 16.5-fold longer than the wild-type enzyme. A comparison of the crystal structures of the full-length wild-type enzyme and three mutants showed that various mechanisms appear to be involved in thermostability enhancement. In particular, the replacement of Phe447 with Val or Pro induced a conformational change in an adjacent residue His477, which results in the formation of a new hydrogen bond in the enzyme. Although the thermostabilization mechanisms of the five residue mutations were explicable on the basis of the crystal structures, it appears to be difficult to predict which amino acid residues should be modified to obtain thermostabilized enzymes. © Springer-Verlag Berlin Heidelberg 2014 |
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Abstract A β-fructofuranosidase from Microbacterium saccharophilum K-1 (formerly known as Arthrobacter sp. K-1) is useful for producing the sweetener lactosucrose ($ 4^{G} $-β-d-galactosylsucrose). Thermostability of the β-fructofuranosidase was enhanced by random mutagenesis and saturation mutagenesis. Clones with enhanced thermostability included mutations at residues Thr47, Ser200, Phe447, Phe470, and Pro500. In the highest stability mutant, T47S/S200T/F447P/F470Y/P500S, the half-life at 60 °C was 182 min, 16.5-fold longer than the wild-type enzyme. A comparison of the crystal structures of the full-length wild-type enzyme and three mutants showed that various mechanisms appear to be involved in thermostability enhancement. In particular, the replacement of Phe447 with Val or Pro induced a conformational change in an adjacent residue His477, which results in the formation of a new hydrogen bond in the enzyme. Although the thermostabilization mechanisms of the five residue mutations were explicable on the basis of the crystal structures, it appears to be difficult to predict which amino acid residues should be modified to obtain thermostabilized enzymes. © Springer-Verlag Berlin Heidelberg 2014 |
| abstract_unstemmed |
Abstract A β-fructofuranosidase from Microbacterium saccharophilum K-1 (formerly known as Arthrobacter sp. K-1) is useful for producing the sweetener lactosucrose ($ 4^{G} $-β-d-galactosylsucrose). Thermostability of the β-fructofuranosidase was enhanced by random mutagenesis and saturation mutagenesis. Clones with enhanced thermostability included mutations at residues Thr47, Ser200, Phe447, Phe470, and Pro500. In the highest stability mutant, T47S/S200T/F447P/F470Y/P500S, the half-life at 60 °C was 182 min, 16.5-fold longer than the wild-type enzyme. A comparison of the crystal structures of the full-length wild-type enzyme and three mutants showed that various mechanisms appear to be involved in thermostability enhancement. In particular, the replacement of Phe447 with Val or Pro induced a conformational change in an adjacent residue His477, which results in the formation of a new hydrogen bond in the enzyme. Although the thermostabilization mechanisms of the five residue mutations were explicable on the basis of the crystal structures, it appears to be difficult to predict which amino acid residues should be modified to obtain thermostabilized enzymes. © Springer-Verlag Berlin Heidelberg 2014 |
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K-1) is useful for producing the sweetener lactosucrose ($ 4^{G} $-β-d-galactosylsucrose). Thermostability of the β-fructofuranosidase was enhanced by random mutagenesis and saturation mutagenesis. Clones with enhanced thermostability included mutations at residues Thr47, Ser200, Phe447, Phe470, and Pro500. In the highest stability mutant, T47S/S200T/F447P/F470Y/P500S, the half-life at 60 °C was 182 min, 16.5-fold longer than the wild-type enzyme. A comparison of the crystal structures of the full-length wild-type enzyme and three mutants showed that various mechanisms appear to be involved in thermostability enhancement. In particular, the replacement of Phe447 with Val or Pro induced a conformational change in an adjacent residue His477, which results in the formation of a new hydrogen bond in the enzyme. 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