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Combinational expression of D-sorbitol dehydrogenase and pyrroloquinoline quinone increases 6-(N-hydroxyethyl)-amino-6-deoxy-α-L-sorbofuranose production by Gluconobacter oxydans through cofactor manipulation.
- Source :
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Enzyme and microbial technology [Enzyme Microb Technol] 2020 Nov; Vol. 141, pp. 109670. Date of Electronic Publication: 2020 Sep 15. - Publication Year :
- 2020
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Abstract
- 6-(N-hydroxyethyl)-amino-6-deoxy-l-sorbofuranose (6NSL), a key precursor in the synthesis of miglitol, is produced from N-2-hydroxyethyl-glucamine (NHEG) by the regioselective oxidation of Gluconobacter oxydans. The limitation of PQQ biosynthesis became a bottleneck for improvement of PQQ-dependent D-sorbitol dehydrogenase (mSLDH) activity. Five expression plasmids were constructed for the co-expression of the pqqABCDE gene cluster and the tldD gene on the basis of pBBR1-gHp0169-sldAB in G. oxydans to increase the biosynthesis of PQQ. The G. oxydans/pGA004, in which pqqABCDE and tldD were expressed as a cluster under the control of gHp0169 promoter, showed the optimal performance. The intracellular PQQ concentration and specific activity of mSLDH in cells increased by 79.3 % and 53.7 %, respectively, compared to that in G. oxydans/pBBR-sldAB. Then, the repeated batch biotransformation of NHEG to 6NSL by G. oxydans/pGA004 was carried out. Up to 75.0 ± 3.0 g/L of 6NSL production with 94.5 ± 3.6 % of average conversion rate of NHEG to 6NSL was achieved after four cycles of run. These results indicated that G. oxydans/pGA004 with high productivity had great potential for 6NSL production in industrial bioprocess.<br /> (Copyright © 2020 Elsevier Inc. All rights reserved.)
- Subjects :
- Bacterial Proteins genetics
Bacterial Proteins metabolism
Bioreactors
Biotransformation
Gene Expression
Gluconobacter oxydans genetics
Gluconobacter oxydans growth & development
L-Iditol 2-Dehydrogenase genetics
Multigene Family
Nitrosamines metabolism
PQQ Cofactor genetics
PQQ Cofactor metabolism
Promoter Regions, Genetic
Recombinant Proteins genetics
Recombinant Proteins metabolism
Sorbose biosynthesis
Gluconobacter oxydans metabolism
L-Iditol 2-Dehydrogenase metabolism
PQQ Cofactor biosynthesis
Sorbose analogs & derivatives
Subjects
Details
- Language :
- English
- ISSN :
- 1879-0909
- Volume :
- 141
- Database :
- MEDLINE
- Journal :
- Enzyme and microbial technology
- Publication Type :
- Academic Journal
- Accession number :
- 33051020
- Full Text :
- https://doi.org/10.1016/j.enzmictec.2020.109670