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Engineering Gluconobacter cerinus CGMCC 1.110 for direct 2-keto-L-gulonic acid production.
- Source :
-
Applied Microbiology & Biotechnology . Jan2023, Vol. 107 Issue 1, p153-162. 10p. - Publication Year :
- 2023
-
Abstract
- Gluconobacter is a potential strain for single-step production of 2-keto-L-gulonic acid (2-KLG), which is the direct precursor of vitamin C. Three dehydrogenases, namely, sorbitol dehydrogenase (SLDH), sorbose dehydrogenase (SDH), and sorbosone dehydrogenase (SNDH), are involved in the production of 2-KLG from D-sorbitol. In the present study, the potential SNDH/SDH gene cluster in the strain Gluconobacter cerinus CGMCC 1.110 was mined by genome analysis, and its function in transforming L-sorbose to 2-KLG was verified. Proteomic analysis showed that the expression level of SNDH/SDH had a great influence on the titer of 2-KLG, and fermentation results showed that SDH was the rate-limiting enzyme. A systematic metabolic engineering process, which was theoretically suitable for increasing the titer of many products involving membrane-bound dehydrogenase from Gluconobacter, was then performed to improve the 2-KLG titer in G. cerinus CGMCC 1.110 from undetectable to 51.9 g/L in a 5-L bioreactor after fermentation optimization. The strategies used in this study may provide a reference for mining other potential applications of Gluconobacter. Key points: • The potential SNDH/SDH gene cluster in G. cerinus CGMCC 1.110 was mined. • A systematic engineering process was performed to improve the titer of 2-KLG. • The 2-KLG titer was successfully increased from undetectable to 51.9 g/L. [ABSTRACT FROM AUTHOR]
- Subjects :
- *SORBITOL
*GENE clusters
*ENGINEERING
*DEHYDROGENASES
*TITERS
*CARBON metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 01757598
- Volume :
- 107
- Issue :
- 1
- Database :
- Academic Search Index
- Journal :
- Applied Microbiology & Biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 160779424
- Full Text :
- https://doi.org/10.1007/s00253-022-12310-5