Reference - Detail
| RRC ID | 35528 |
|---|---|
| Author | Ashok S, Mohan Raj S, Ko Y, Sankaranarayanan M, Zhou S, Kumar V, Park S. |
| Title | Effect of puuC overexpression and nitrate addition on glycerol metabolism and anaerobic 3-hydroxypropionic acid production in recombinant Klebsiella pneumoniae ΔglpKΔdhaT. |
| Journal | Metab Eng |
| Abstract |
3-Hydroxypropionic acid (3-HP), an industrially important platform chemical, is used as a precursor during the production of many commercially important chemicals. Recently, recombinant strains of K. pneumoniae overexpressing an NAD(+)-dependent γ-glutamyl-γ-aminobutyraldehyde dehydrogenase (PuuC) enzyme of K. pneumoniae DSM 2026 were shown to produce 3-HP from glycerol without the addition coenzyme B(12), which is expensive. However, 3-HP production in K. pneumoniae is accompanied with NADH generation, and this always results in large accumulation of 1,3-propanediol (1,3-PDO) and lactic acid. In this study, we investigated the potential use of nitrate as an electron acceptor both to regenerate NAD(+) and to prevent the formation of byproducts during anaerobic production of 3-HP from glycerol. Nitrate addition could improve NAD(+) regeneration, but decreased glycerol flux towards 3-HP production. To divert more glycerol towards 3-HP, a novel recombinant strain K. pneumoniae ΔglpKΔdhaT (puuC) was developed by disrupting the glpK gene, which encodes glycerol kinase, and the dhaT gene, which encodes 1,3-propanediol oxidoreductase. This strain showed improved cellular NAD(+) concentrations and a high carbon flux towards 3-HP production. Through anaerobic cultivation in the presence of nitrate, this recombinant strain produced more than 40±3mM 3-HP with more than 50% yield on glycerol in shake flasks and 250±10mM 3-HP with approximately 30% yield on glycerol in a fed-batch bioreactor. |
| Volume | 15 |
| Pages | 10-24 |
| Published | 2013-1-1 |
| DOI | 10.1016/j.ymben.2012.09.004 |
| PII | S1096-7176(12)00099-7 |
| PMID | 23022570 |
| MeSH | Aldehyde Oxidoreductases / genetics Aldehyde Oxidoreductases / metabolism* Anaerobiosis / physiology Genetic Enhancement / methods* Glycerol / metabolism* Klebsiella pneumoniae / classification Klebsiella pneumoniae / physiology* Lactic Acid / analogs & derivatives* Lactic Acid / biosynthesis Nitrates / metabolism* Recombination, Genetic / genetics* Species Specificity Up-Regulation |
| IF | 7.263 |
| Times Cited | 58 |
| WOS Category | BIOTECHNOLOGY & APPLIED MICROBIOLOGY |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | Patent(IFI CLAIMS) |
| Total number of mentions | 5 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Prokaryotes E. coli | NA |