Regulation of Pyruvate Formate Lyase-Deficient Klebsiella pneumoniae for Efficient 1,3-Propanediol Bioproduction
Anaerobic growth defect of pyruvate formate lyase (PFL)-deficient Klebsiella pneumoniae limits its industrial application, and the reason for this growth defect was analyzed in this study. The obtained evidences, combined with normal intracellular redox status and no further inhibition by adhE deletion, strongly suggested that growth defect in PFL-deficient K. pneumoniae was probably caused by lack of carbon flux from pyruvate to acetyl-CoA (AcCoA). Correspondingly, the anaerobic growth of PFL-deficient K. pneumoniae was promoted by deletion of pdhR, a negative transcriptional regulator gene for AcCoA generation. Through the regulation of pdhR deletion, the PFL-deficient K. pneumoniae exhibited highly efficient 1,3-propanediol production. Besides, in a 2-L fed-batch fermentation process, the cell growth of PFL-deficient K. pneumoniae strain almost recovered, when compared with that of the normal strain, and the 1,3-propanediol yield increased by 14%, while the byproducts acetate and 2,3-butanediol contents decreased by 29% and 24%, respectively.
This work was supported by the National Natural Science Foundation of China under Grant No. 31271862.
- 5.Kumar V, Sankaranarayanan M, Jae KE, Durgapal M, Ashok S, Ko Y, Sarkar R, Park S (2012) Co-production of 3-hydroxypropionic acid and 1,3-propanediol from glycerol using resting cells of recombinant Klebsiella pneumoniae J2B strain overexpressing aldehyde dehydrogenase. Appl Microbiol Biotechnol 96(2):373–383. https://doi.org/10.1007/s00253-012-4187-9 CrossRefPubMedGoogle Scholar
- 6.Zhuge B, Zhang C, Fang H, Zhuge J, Permaul K (2010) Expression of 1,3-propanediol oxidoreductase and its isoenzyme in Klebsiella pneumoniae for bioconversion of glycerol into 1,3-propanediol. Appl Microbiol Biotechnol 87(6):2177–2184. https://doi.org/10.1007/s00253-010-2678-0 CrossRefPubMedGoogle Scholar
- 7.Kumar V, Durgapal M, Sankaranarayanan M, Somasundar A, Rathnasingh C, Song H, Seung D, Park S (2016) Effects of mutation of 2,3-butanediol formation pathway on glycerol metabolism and 1,3-propanediol production by Klebsiella pneumoniae J2B. Bioresour Technol 214:432–440. https://doi.org/10.1016/j.biortech.2016.04.032 CrossRefPubMedGoogle Scholar
- 9.Ko Y, Seol E, Sundara Sekar B, Kwon S, Lee J, Park S (2017) Metabolic engineering of Klebsiella pneumoniae J2B for co-production of 3-hydroxypropionic acid and 1,3-propanediol from glycerol: reduction of acetate and other by-products. Bioresour Technol 244(Pt 1):1096–1103. https://doi.org/10.1016/j.biortech.2017.08.099 CrossRefPubMedGoogle Scholar
- 17.Murarka A, Clomburg JM, Moran S, Shanks JV, Gonzalez R (2010) Metabolic analysis of wild-type Escherichia coli and a pyruvate dehydrogenase complex (PDHC)-deficient derivative reveals the role of PDHC in the fermentative metabolism of glucose. J Biol Chem 285(41):31548–31558. https://doi.org/10.1074/jbc.M110.121095 CrossRefPubMedPubMedCentralGoogle Scholar
- 24.Du C, Zhang Y, Li Y, Cao Z (2007) Novel redox potential-based screening strategy for rapid isolation of Klebsiella pneumoniae mutants with enhanced 1,3-propanediol-producing capability. Appl Environ Microbiol 73(14):4515–4521. https://doi.org/10.1128/AEM.02857-06 CrossRefPubMedPubMedCentralGoogle Scholar
- 26.Maeda S, Shimizu K, Kihira C, Iwabu Y, Kato R, Sugimoto M, Fukiya S, Wada M, Yokota A (2017) Pyruvate dehydrogenase complex regulator (PdhR) gene deletion boosts glucose metabolism in Escherichia coli under oxygen-limited culture conditions. J Biosci Bioeng 123(4):437–443. https://doi.org/10.1016/j.jbiosc.2016.11.004 CrossRefPubMedGoogle Scholar