Breeding of

6-(N-hydroxyethyl)-amino-6-deoxy-L-sorbofuranose (6NSL), a key intermediate in the synthesis of miglitol, was produced from N-2-hydroxyethyl glucamine (NHEG) by biotransformation with whole cells of Gluconobacter oxydans. The main troubles in 6NSL production were the low activity of PQQ-dependent D-...
Ausführliche Beschreibung

Gespeichert in:

Autor*in:	Liu, Dong [verfasserIn] Hu, Zhong-Ce [verfasserIn] Ke, Xia [verfasserIn] Zheng, Yu-Guo [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2020

Schlagwörter:	6-( D-Sorbitol dehydrogenase (mSLDH) High-throughput screening method Combined mutagenesis

Übergeordnetes Werk:	Enthalten in: Biochemical engineering journal - Amsterdam [u.a.] : Elsevier, 1998, 161
Übergeordnetes Werk:	volume:161

DOI / URN:	10.1016/j.bej.2020.107642

Katalog-ID:	ELV004412109

Internformat


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520			\|a 6-(N-hydroxyethyl)-amino-6-deoxy-L-sorbofuranose (6NSL), a key intermediate in the synthesis of miglitol, was produced from N-2-hydroxyethyl glucamine (NHEG) by biotransformation with whole cells of Gluconobacter oxydans. The main troubles in 6NSL production were the low activity of PQQ-dependent D-sorbitol dehydrogenase (mSLDH) in G. oxydans and the high cost of cell preparation. To solve these problems, a combined mutagenesis of 60Co-γ irradiation and microwave treatment with a high-throughput screening method by cultivation in a 96-well microtiter plate was conducted. After several cycles of mutagenesis, a stable mutant H-8 with high mSLDH activity was obtained, and the cell biomass increased by 11.6% when cultivated in a 5 L bioreactor. The transcription levels of the mSLDH subunit sldA and sldB in G. oxydans H-8 increased by 1.4- and 2.0-fold, respectively. Meanwhile, the intracellular PQQ concentration in G. oxydans H-8 was 16.1% higher than that of the parent strain, and qRT-PCR analysis showed that the genes pqqB and pqqC played an important role in PQQ biosynthesis (transcription levels increased by 2.4- and 1.8-fold, respectively). Relying on the advantages of the above, G. oxydans H-8 could produce 64.3 ± 2.2 g/L 6NSL after 36 h of bioconversion with resting cells, showing a 33.7% increase in the product yield.
650		4	\|a 6-(
650		4	\|a D-Sorbitol dehydrogenase (mSLDH)
650		4	\|a High-throughput screening method
650		4	\|a Combined mutagenesis
700	1		\|a Hu, Zhong-Ce \|e verfasserin \|0 (orcid)0000-0002-5849-2744 \|4 aut
700	1		\|a Ke, Xia \|e verfasserin \|4 aut
700	1		\|a Zheng, Yu-Guo \|e verfasserin \|4 aut
773	0	8	\|i Enthalten in \|t Biochemical engineering journal \|d Amsterdam [u.a.] : Elsevier, 1998 \|g 161 \|h Online-Ressource \|w (DE-627)320500349 \|w (DE-600)2012139-8 \|w (DE-576)098330160 \|7 nnns
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912			\|a GBV_USEFLAG_U
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912			\|a GBV_ILN_31
912			\|a GBV_ILN_32
912			\|a GBV_ILN_40
912			\|a GBV_ILN_60
912			\|a GBV_ILN_62
912			\|a GBV_ILN_63
912			\|a GBV_ILN_65
912			\|a GBV_ILN_69
912			\|a GBV_ILN_70
912			\|a GBV_ILN_73
912			\|a GBV_ILN_74
912			\|a GBV_ILN_90
912			\|a GBV_ILN_95
912			\|a GBV_ILN_100
912			\|a GBV_ILN_101
912			\|a GBV_ILN_105
912			\|a GBV_ILN_110
912			\|a GBV_ILN_150
912			\|a GBV_ILN_151
912			\|a GBV_ILN_224
912			\|a GBV_ILN_370
912			\|a GBV_ILN_602
912			\|a GBV_ILN_702
912			\|a GBV_ILN_2003
912			\|a GBV_ILN_2004
912			\|a GBV_ILN_2005
912			\|a GBV_ILN_2011
912			\|a GBV_ILN_2014
912			\|a GBV_ILN_2015
912			\|a GBV_ILN_2020
912			\|a GBV_ILN_2021
912			\|a GBV_ILN_2025
912			\|a GBV_ILN_2027
912			\|a GBV_ILN_2034
912			\|a GBV_ILN_2038
912			\|a GBV_ILN_2044
912			\|a GBV_ILN_2048
912			\|a GBV_ILN_2049
912			\|a GBV_ILN_2050
912			\|a GBV_ILN_2056
912			\|a GBV_ILN_2059
912			\|a GBV_ILN_2061
912			\|a GBV_ILN_2064
912			\|a GBV_ILN_2065
912			\|a GBV_ILN_2068
912			\|a GBV_ILN_2111
912			\|a GBV_ILN_2112
912			\|a GBV_ILN_2113
912			\|a GBV_ILN_2118
912			\|a GBV_ILN_2122
912			\|a GBV_ILN_2129
912			\|a GBV_ILN_2143
912			\|a GBV_ILN_2147
912			\|a GBV_ILN_2148
912			\|a GBV_ILN_2152
912			\|a GBV_ILN_2153
912			\|a GBV_ILN_2190
912			\|a GBV_ILN_2336
912			\|a GBV_ILN_2507
912			\|a GBV_ILN_2522
912			\|a GBV_ILN_4035
912			\|a GBV_ILN_4037
912			\|a GBV_ILN_4112
912			\|a GBV_ILN_4125
912			\|a GBV_ILN_4126
912			\|a GBV_ILN_4242
912			\|a GBV_ILN_4251
912			\|a GBV_ILN_4305
912			\|a GBV_ILN_4313
912			\|a GBV_ILN_4323
912			\|a GBV_ILN_4324
912			\|a GBV_ILN_4326
912			\|a GBV_ILN_4333
912			\|a GBV_ILN_4334
912			\|a GBV_ILN_4335
912			\|a GBV_ILN_4338
912			\|a GBV_ILN_4393
936	b	k	\|a 58.30 \|j Biotechnologie
936	b	k	\|a 58.00 \|j Chemische Technik: Allgemeines
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author_variant	d l dl z c h zch x k xk y g z ygz
matchkey_str	liudonghuzhongcekexiazhengyuguo:2020----:rei
hierarchy_sort_str	2020
bklnumber	58.30 58.00
publishDate	2020
allfields	10.1016/j.bej.2020.107642 doi (DE-627)ELV004412109 (ELSEVIER)S1369-703X(20)30196-0 DE-627 ger DE-627 rda eng 660 540 DE-600 58.30 bkl 58.00 bkl Liu, Dong verfasserin (orcid)0000-0002-6967-3348 aut Breeding of 2020 nicht spezifiziert zzz rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 6-(N-hydroxyethyl)-amino-6-deoxy-L-sorbofuranose (6NSL), a key intermediate in the synthesis of miglitol, was produced from N-2-hydroxyethyl glucamine (NHEG) by biotransformation with whole cells of Gluconobacter oxydans. The main troubles in 6NSL production were the low activity of PQQ-dependent D-sorbitol dehydrogenase (mSLDH) in G. oxydans and the high cost of cell preparation. To solve these problems, a combined mutagenesis of 60Co-γ irradiation and microwave treatment with a high-throughput screening method by cultivation in a 96-well microtiter plate was conducted. After several cycles of mutagenesis, a stable mutant H-8 with high mSLDH activity was obtained, and the cell biomass increased by 11.6% when cultivated in a 5 L bioreactor. The transcription levels of the mSLDH subunit sldA and sldB in G. oxydans H-8 increased by 1.4- and 2.0-fold, respectively. Meanwhile, the intracellular PQQ concentration in G. oxydans H-8 was 16.1% higher than that of the parent strain, and qRT-PCR analysis showed that the genes pqqB and pqqC played an important role in PQQ biosynthesis (transcription levels increased by 2.4- and 1.8-fold, respectively). Relying on the advantages of the above, G. oxydans H-8 could produce 64.3 ± 2.2 g/L 6NSL after 36 h of bioconversion with resting cells, showing a 33.7% increase in the product yield. 6-( D-Sorbitol dehydrogenase (mSLDH) High-throughput screening method Combined mutagenesis Hu, Zhong-Ce verfasserin (orcid)0000-0002-5849-2744 aut Ke, Xia verfasserin aut Zheng, Yu-Guo verfasserin aut Enthalten in Biochemical engineering journal Amsterdam [u.a.] : Elsevier, 1998 161 Online-Ressource (DE-627)320500349 (DE-600)2012139-8 (DE-576)098330160 nnns volume:161 GBV_USEFLAG_U SYSFLAG_U GBV_ELV SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_110 GBV_ILN_150 GBV_ILN_151 GBV_ILN_224 GBV_ILN_370 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2027 GBV_ILN_2034 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2056 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2190 GBV_ILN_2336 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4313 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4393 58.30 Biotechnologie 58.00 Chemische Technik: Allgemeines AR 161
spelling	10.1016/j.bej.2020.107642 doi (DE-627)ELV004412109 (ELSEVIER)S1369-703X(20)30196-0 DE-627 ger DE-627 rda eng 660 540 DE-600 58.30 bkl 58.00 bkl Liu, Dong verfasserin (orcid)0000-0002-6967-3348 aut Breeding of 2020 nicht spezifiziert zzz rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 6-(N-hydroxyethyl)-amino-6-deoxy-L-sorbofuranose (6NSL), a key intermediate in the synthesis of miglitol, was produced from N-2-hydroxyethyl glucamine (NHEG) by biotransformation with whole cells of Gluconobacter oxydans. The main troubles in 6NSL production were the low activity of PQQ-dependent D-sorbitol dehydrogenase (mSLDH) in G. oxydans and the high cost of cell preparation. To solve these problems, a combined mutagenesis of 60Co-γ irradiation and microwave treatment with a high-throughput screening method by cultivation in a 96-well microtiter plate was conducted. After several cycles of mutagenesis, a stable mutant H-8 with high mSLDH activity was obtained, and the cell biomass increased by 11.6% when cultivated in a 5 L bioreactor. The transcription levels of the mSLDH subunit sldA and sldB in G. oxydans H-8 increased by 1.4- and 2.0-fold, respectively. Meanwhile, the intracellular PQQ concentration in G. oxydans H-8 was 16.1% higher than that of the parent strain, and qRT-PCR analysis showed that the genes pqqB and pqqC played an important role in PQQ biosynthesis (transcription levels increased by 2.4- and 1.8-fold, respectively). Relying on the advantages of the above, G. oxydans H-8 could produce 64.3 ± 2.2 g/L 6NSL after 36 h of bioconversion with resting cells, showing a 33.7% increase in the product yield. 6-( D-Sorbitol dehydrogenase (mSLDH) High-throughput screening method Combined mutagenesis Hu, Zhong-Ce verfasserin (orcid)0000-0002-5849-2744 aut Ke, Xia verfasserin aut Zheng, Yu-Guo verfasserin aut Enthalten in Biochemical engineering journal Amsterdam [u.a.] : Elsevier, 1998 161 Online-Ressource (DE-627)320500349 (DE-600)2012139-8 (DE-576)098330160 nnns volume:161 GBV_USEFLAG_U SYSFLAG_U GBV_ELV SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_110 GBV_ILN_150 GBV_ILN_151 GBV_ILN_224 GBV_ILN_370 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2027 GBV_ILN_2034 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2056 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2190 GBV_ILN_2336 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4313 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4393 58.30 Biotechnologie 58.00 Chemische Technik: Allgemeines AR 161
allfields_unstemmed	10.1016/j.bej.2020.107642 doi (DE-627)ELV004412109 (ELSEVIER)S1369-703X(20)30196-0 DE-627 ger DE-627 rda eng 660 540 DE-600 58.30 bkl 58.00 bkl Liu, Dong verfasserin (orcid)0000-0002-6967-3348 aut Breeding of 2020 nicht spezifiziert zzz rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 6-(N-hydroxyethyl)-amino-6-deoxy-L-sorbofuranose (6NSL), a key intermediate in the synthesis of miglitol, was produced from N-2-hydroxyethyl glucamine (NHEG) by biotransformation with whole cells of Gluconobacter oxydans. The main troubles in 6NSL production were the low activity of PQQ-dependent D-sorbitol dehydrogenase (mSLDH) in G. oxydans and the high cost of cell preparation. To solve these problems, a combined mutagenesis of 60Co-γ irradiation and microwave treatment with a high-throughput screening method by cultivation in a 96-well microtiter plate was conducted. After several cycles of mutagenesis, a stable mutant H-8 with high mSLDH activity was obtained, and the cell biomass increased by 11.6% when cultivated in a 5 L bioreactor. The transcription levels of the mSLDH subunit sldA and sldB in G. oxydans H-8 increased by 1.4- and 2.0-fold, respectively. Meanwhile, the intracellular PQQ concentration in G. oxydans H-8 was 16.1% higher than that of the parent strain, and qRT-PCR analysis showed that the genes pqqB and pqqC played an important role in PQQ biosynthesis (transcription levels increased by 2.4- and 1.8-fold, respectively). Relying on the advantages of the above, G. oxydans H-8 could produce 64.3 ± 2.2 g/L 6NSL after 36 h of bioconversion with resting cells, showing a 33.7% increase in the product yield. 6-( D-Sorbitol dehydrogenase (mSLDH) High-throughput screening method Combined mutagenesis Hu, Zhong-Ce verfasserin (orcid)0000-0002-5849-2744 aut Ke, Xia verfasserin aut Zheng, Yu-Guo verfasserin aut Enthalten in Biochemical engineering journal Amsterdam [u.a.] : Elsevier, 1998 161 Online-Ressource (DE-627)320500349 (DE-600)2012139-8 (DE-576)098330160 nnns volume:161 GBV_USEFLAG_U SYSFLAG_U GBV_ELV SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_110 GBV_ILN_150 GBV_ILN_151 GBV_ILN_224 GBV_ILN_370 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2027 GBV_ILN_2034 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2056 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2190 GBV_ILN_2336 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4313 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4393 58.30 Biotechnologie 58.00 Chemische Technik: Allgemeines AR 161
allfieldsGer	10.1016/j.bej.2020.107642 doi (DE-627)ELV004412109 (ELSEVIER)S1369-703X(20)30196-0 DE-627 ger DE-627 rda eng 660 540 DE-600 58.30 bkl 58.00 bkl Liu, Dong verfasserin (orcid)0000-0002-6967-3348 aut Breeding of 2020 nicht spezifiziert zzz rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 6-(N-hydroxyethyl)-amino-6-deoxy-L-sorbofuranose (6NSL), a key intermediate in the synthesis of miglitol, was produced from N-2-hydroxyethyl glucamine (NHEG) by biotransformation with whole cells of Gluconobacter oxydans. The main troubles in 6NSL production were the low activity of PQQ-dependent D-sorbitol dehydrogenase (mSLDH) in G. oxydans and the high cost of cell preparation. To solve these problems, a combined mutagenesis of 60Co-γ irradiation and microwave treatment with a high-throughput screening method by cultivation in a 96-well microtiter plate was conducted. After several cycles of mutagenesis, a stable mutant H-8 with high mSLDH activity was obtained, and the cell biomass increased by 11.6% when cultivated in a 5 L bioreactor. The transcription levels of the mSLDH subunit sldA and sldB in G. oxydans H-8 increased by 1.4- and 2.0-fold, respectively. Meanwhile, the intracellular PQQ concentration in G. oxydans H-8 was 16.1% higher than that of the parent strain, and qRT-PCR analysis showed that the genes pqqB and pqqC played an important role in PQQ biosynthesis (transcription levels increased by 2.4- and 1.8-fold, respectively). Relying on the advantages of the above, G. oxydans H-8 could produce 64.3 ± 2.2 g/L 6NSL after 36 h of bioconversion with resting cells, showing a 33.7% increase in the product yield. 6-( D-Sorbitol dehydrogenase (mSLDH) High-throughput screening method Combined mutagenesis Hu, Zhong-Ce verfasserin (orcid)0000-0002-5849-2744 aut Ke, Xia verfasserin aut Zheng, Yu-Guo verfasserin aut Enthalten in Biochemical engineering journal Amsterdam [u.a.] : Elsevier, 1998 161 Online-Ressource (DE-627)320500349 (DE-600)2012139-8 (DE-576)098330160 nnns volume:161 GBV_USEFLAG_U SYSFLAG_U GBV_ELV SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_110 GBV_ILN_150 GBV_ILN_151 GBV_ILN_224 GBV_ILN_370 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2027 GBV_ILN_2034 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2056 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2190 GBV_ILN_2336 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4313 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4393 58.30 Biotechnologie 58.00 Chemische Technik: Allgemeines AR 161
allfieldsSound	10.1016/j.bej.2020.107642 doi (DE-627)ELV004412109 (ELSEVIER)S1369-703X(20)30196-0 DE-627 ger DE-627 rda eng 660 540 DE-600 58.30 bkl 58.00 bkl Liu, Dong verfasserin (orcid)0000-0002-6967-3348 aut Breeding of 2020 nicht spezifiziert zzz rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 6-(N-hydroxyethyl)-amino-6-deoxy-L-sorbofuranose (6NSL), a key intermediate in the synthesis of miglitol, was produced from N-2-hydroxyethyl glucamine (NHEG) by biotransformation with whole cells of Gluconobacter oxydans. The main troubles in 6NSL production were the low activity of PQQ-dependent D-sorbitol dehydrogenase (mSLDH) in G. oxydans and the high cost of cell preparation. To solve these problems, a combined mutagenesis of 60Co-γ irradiation and microwave treatment with a high-throughput screening method by cultivation in a 96-well microtiter plate was conducted. After several cycles of mutagenesis, a stable mutant H-8 with high mSLDH activity was obtained, and the cell biomass increased by 11.6% when cultivated in a 5 L bioreactor. The transcription levels of the mSLDH subunit sldA and sldB in G. oxydans H-8 increased by 1.4- and 2.0-fold, respectively. Meanwhile, the intracellular PQQ concentration in G. oxydans H-8 was 16.1% higher than that of the parent strain, and qRT-PCR analysis showed that the genes pqqB and pqqC played an important role in PQQ biosynthesis (transcription levels increased by 2.4- and 1.8-fold, respectively). Relying on the advantages of the above, G. oxydans H-8 could produce 64.3 ± 2.2 g/L 6NSL after 36 h of bioconversion with resting cells, showing a 33.7% increase in the product yield. 6-( D-Sorbitol dehydrogenase (mSLDH) High-throughput screening method Combined mutagenesis Hu, Zhong-Ce verfasserin (orcid)0000-0002-5849-2744 aut Ke, Xia verfasserin aut Zheng, Yu-Guo verfasserin aut Enthalten in Biochemical engineering journal Amsterdam [u.a.] : Elsevier, 1998 161 Online-Ressource (DE-627)320500349 (DE-600)2012139-8 (DE-576)098330160 nnns volume:161 GBV_USEFLAG_U SYSFLAG_U GBV_ELV SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_110 GBV_ILN_150 GBV_ILN_151 GBV_ILN_224 GBV_ILN_370 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2027 GBV_ILN_2034 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2056 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2190 GBV_ILN_2336 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4313 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4393 58.30 Biotechnologie 58.00 Chemische Technik: Allgemeines AR 161
language	English
source	Enthalten in Biochemical engineering journal 161 volume:161
sourceStr	Enthalten in Biochemical engineering journal 161 volume:161
format_phy_str_mv	Article
bklname	Biotechnologie Chemische Technik: Allgemeines
institution	findex.gbv.de
topic_facet	6-( D-Sorbitol dehydrogenase (mSLDH) High-throughput screening method Combined mutagenesis
dewey-raw	660
isfreeaccess_bool	false
container_title	Biochemical engineering journal
authorswithroles_txt_mv	Liu, Dong @@aut@@ Hu, Zhong-Ce @@aut@@ Ke, Xia @@aut@@ Zheng, Yu-Guo @@aut@@
publishDateDaySort_date	2020-01-01T00:00:00Z
hierarchy_top_id	320500349
dewey-sort	3660
id	ELV004412109
language_de	englisch
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author	Liu, Dong
spellingShingle	Liu, Dong ddc 660 bkl 58.30 bkl 58.00 misc 6-( misc D-Sorbitol dehydrogenase (mSLDH) misc High-throughput screening method misc Combined mutagenesis Breeding of
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topic_title	660 540 DE-600 58.30 bkl 58.00 bkl Breeding of 6-( D-Sorbitol dehydrogenase (mSLDH) High-throughput screening method Combined mutagenesis
topic	ddc 660 bkl 58.30 bkl 58.00 misc 6-( misc D-Sorbitol dehydrogenase (mSLDH) misc High-throughput screening method misc Combined mutagenesis
topic_unstemmed	ddc 660 bkl 58.30 bkl 58.00 misc 6-( misc D-Sorbitol dehydrogenase (mSLDH) misc High-throughput screening method misc Combined mutagenesis
topic_browse	ddc 660 bkl 58.30 bkl 58.00 misc 6-( misc D-Sorbitol dehydrogenase (mSLDH) misc High-throughput screening method misc Combined mutagenesis
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title	Breeding of
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title_full	Breeding of
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title_sort	breeding of
title_auth	Breeding of
abstract	6-(N-hydroxyethyl)-amino-6-deoxy-L-sorbofuranose (6NSL), a key intermediate in the synthesis of miglitol, was produced from N-2-hydroxyethyl glucamine (NHEG) by biotransformation with whole cells of Gluconobacter oxydans. The main troubles in 6NSL production were the low activity of PQQ-dependent D-sorbitol dehydrogenase (mSLDH) in G. oxydans and the high cost of cell preparation. To solve these problems, a combined mutagenesis of 60Co-γ irradiation and microwave treatment with a high-throughput screening method by cultivation in a 96-well microtiter plate was conducted. After several cycles of mutagenesis, a stable mutant H-8 with high mSLDH activity was obtained, and the cell biomass increased by 11.6% when cultivated in a 5 L bioreactor. The transcription levels of the mSLDH subunit sldA and sldB in G. oxydans H-8 increased by 1.4- and 2.0-fold, respectively. Meanwhile, the intracellular PQQ concentration in G. oxydans H-8 was 16.1% higher than that of the parent strain, and qRT-PCR analysis showed that the genes pqqB and pqqC played an important role in PQQ biosynthesis (transcription levels increased by 2.4- and 1.8-fold, respectively). Relying on the advantages of the above, G. oxydans H-8 could produce 64.3 ± 2.2 g/L 6NSL after 36 h of bioconversion with resting cells, showing a 33.7% increase in the product yield.
abstractGer	6-(N-hydroxyethyl)-amino-6-deoxy-L-sorbofuranose (6NSL), a key intermediate in the synthesis of miglitol, was produced from N-2-hydroxyethyl glucamine (NHEG) by biotransformation with whole cells of Gluconobacter oxydans. The main troubles in 6NSL production were the low activity of PQQ-dependent D-sorbitol dehydrogenase (mSLDH) in G. oxydans and the high cost of cell preparation. To solve these problems, a combined mutagenesis of 60Co-γ irradiation and microwave treatment with a high-throughput screening method by cultivation in a 96-well microtiter plate was conducted. After several cycles of mutagenesis, a stable mutant H-8 with high mSLDH activity was obtained, and the cell biomass increased by 11.6% when cultivated in a 5 L bioreactor. The transcription levels of the mSLDH subunit sldA and sldB in G. oxydans H-8 increased by 1.4- and 2.0-fold, respectively. Meanwhile, the intracellular PQQ concentration in G. oxydans H-8 was 16.1% higher than that of the parent strain, and qRT-PCR analysis showed that the genes pqqB and pqqC played an important role in PQQ biosynthesis (transcription levels increased by 2.4- and 1.8-fold, respectively). Relying on the advantages of the above, G. oxydans H-8 could produce 64.3 ± 2.2 g/L 6NSL after 36 h of bioconversion with resting cells, showing a 33.7% increase in the product yield.
abstract_unstemmed	6-(N-hydroxyethyl)-amino-6-deoxy-L-sorbofuranose (6NSL), a key intermediate in the synthesis of miglitol, was produced from N-2-hydroxyethyl glucamine (NHEG) by biotransformation with whole cells of Gluconobacter oxydans. The main troubles in 6NSL production were the low activity of PQQ-dependent D-sorbitol dehydrogenase (mSLDH) in G. oxydans and the high cost of cell preparation. To solve these problems, a combined mutagenesis of 60Co-γ irradiation and microwave treatment with a high-throughput screening method by cultivation in a 96-well microtiter plate was conducted. After several cycles of mutagenesis, a stable mutant H-8 with high mSLDH activity was obtained, and the cell biomass increased by 11.6% when cultivated in a 5 L bioreactor. The transcription levels of the mSLDH subunit sldA and sldB in G. oxydans H-8 increased by 1.4- and 2.0-fold, respectively. Meanwhile, the intracellular PQQ concentration in G. oxydans H-8 was 16.1% higher than that of the parent strain, and qRT-PCR analysis showed that the genes pqqB and pqqC played an important role in PQQ biosynthesis (transcription levels increased by 2.4- and 1.8-fold, respectively). Relying on the advantages of the above, G. oxydans H-8 could produce 64.3 ± 2.2 g/L 6NSL after 36 h of bioconversion with resting cells, showing a 33.7% increase in the product yield.
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title_short	Breeding of
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author2	Hu, Zhong-Ce Ke, Xia Zheng, Yu-Guo
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doi_str	10.1016/j.bej.2020.107642
up_date	2024-07-06T22:54:10.058Z
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score	7.400962

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