Dataset Information

High-yield production of 1,3-propanediol from glycerol by metabolically engineered Klebsiella pneumoniae.

ABSTRACT: Background:Glycerol is a major byproduct of the biodiesel industry and can be converted to 1,3-propanediol (1,3-PDO) by microorganisms through a two-step enzymatic reaction. The production of 1,3-PDO from glycerol using microorganisms is accompanied by formation of unwanted byproducts, including lactate and 2,3-butanediol, resulting in a low-conversion yield. Results:Klebsiella pneumoniae was metabolically engineered to produce high-molar yield of 1,3-PDO from glycerol. First, the pathway genes for byproduct formation were deleted in K. pneumoniae. Then, glycerol assimilation pathways were eliminated and mannitol was co-fed to the medium. Finally, transcriptional regulation of the dha operon were genetically modified for enhancing 1,3-propanediol production. The batch fermentation of the engineered strain with co-feeding of a small amount of mannitol yielded 0.76 mol 1,3-PDO from 1 mol glycerol. Conclusions:Klebsiella pneumoniae is useful microorganism for producing 1,3-PDO from glycerol. Implemented engineering in this study successfully improved 1,3-PDO production yield, which is significantly higher than those reported in previous studies.

SUBMITTER: Lee JH

PROVIDER: S-EPMC5890353 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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High-yield production of 1,3-propanediol from glycerol by metabolically engineered <i>Klebsiella pneumoniae</i>.

Lee Jung Hun JH Jung Moo-Young MY Oh Min-Kyu MK

Biotechnology for biofuels 20180409

<h4>Background</h4>Glycerol is a major byproduct of the biodiesel industry and can be converted to 1,3-propanediol (1,3-PDO) by microorganisms through a two-step enzymatic reaction. The production of 1,3-PDO from glycerol using microorganisms is accompanied by formation of unwanted byproducts, including lactate and 2,3-butanediol, resulting in a low-conversion yield.<h4>Results</h4><i>Klebsiella pneumoniae</i> was metabolically engineered to produce high-molar yield of 1,3-PDO from glycerol. Fir ...[more]

PMID: 29657579

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Project description:BACKGROUND:2,3-Butanediol (2,3-BDO) is a promising bio-based chemical because of its wide industrial applications. Previous studies on microbial production of 2,3-BDO has focused on sugar fermentation. Alternatively, biodiesel-derived crude glycerol can be used as a cheap resource for 2,3-BDO production; however, a considerable formation of 1,3-propanediol (1,3-PDO) and low concentration, productivity, and yield of 2,3-BDO from glycerol fermentation are limitations. RESULTS:Here, we report a high production of 2,3-BDO from crude glycerol using the engineered Klebsiella oxytoca M3 in which pduC (encoding glycerol dehydratase large subunit) and ldhA (encoding lactate dehydrogenase) were deleted to reduce the formation of 1,3-PDO and lactic acid. In fed-batch fermentation with the parent strain K. oxytoca M1, crude glycerol was more effective than pure glycerol as a carbon source in 2,3-BDO production (59.4 vs. 73.8 g/L) and by-product reduction (1,3-PDO, 8.9 vs. 3.7 g/L; lactic acid, 18.6 vs. 9.8 g/L). When the double mutant was used in fed-batch fermentation with pure glycerol, cell growth and glycerol consumption were significantly enhanced and 2,3-BDO production was 1.9-fold higher than that of the parent strain (59.4 vs. 115.0 g/L) with 6.9 g/L of 1,3-PDO and a small amount of lactic acid (0.7 g/L). Notably, when crude glycerol was supplied, the double mutant showed 1,3-PDO-free 2,3-BDO production with high concentration (131.5 g/L), productivity (0.84 g/L/h), and yield (0.44 g/g crude glycerol). This result is the highest 2,3-BDO production from glycerol fermentation to date. CONCLUSIONS:2,3-BDO production from glycerol was dramatically enhanced by disruption of the pduC and ldhA genes in K. oxytoca M1 and 1,3-PDO-free 2,3-BDO production was achieved by using the double mutant and crude glycerol. 2,3-BDO production obtained in this study is comparable to 2,3-BDO production from sugar fermentation, demonstrating the feasibility of economic industrial 2,3-BDO production using crude glycerol.

Project description:Crude glycerol is largely generated as the main by-product of the biodiesel industry and is unprofitable for industrial application without costly purification. The direct bioconversion of crude glycerol into 1,3-propanediol (1,3-PDO) by microorganisms is a promising alternative for effective and economic utilization. In this study, Klebsiella pneumoniae 2e was newly isolated for the conversion of crude glycerol into 1,3-PDO. Batch fermentation analysis confirmed that crude glycerol and its main impurities had slight impacts on the growth, key enzyme activity, and 1,3-PDO production of K. pneumoniae 2e. The 1,3-PDO yield from crude glycerol by K. pneumoniae 2e reached 0.64 mol 1,3-PDO/mol glycerol, which was higher than that by most reported 1,3-PDO-producing Klebsiella strains. Genomic profiling revealed that K. pneumoniae 2e possesses 30 genes involved in glycerol anaerobic metabolism and 1,3-PDO biosynthesis. Quantitative real-time PCR analysis of these genes showed that the majority of the genes encoding the key enzymes for glycerol metabolism and 1,3-PDO biosynthesis were significantly upregulated during culture in crude glycerol relative to that in pure glycerol. Further comparative genomic analysis revealed a novel glycerol uptake facilitator protein in K. pneumoniae 2e and a higher number of stress response proteins than in other Klebsiella strains. This work confirms the adaptability of a newly isolated 1,3-PDO-producing strain, K. pneumoniae 2e, to crude glycerol and provides insights into the molecular mechanisms involved in its crude glycerol tolerance, which is valuable for industrial 1,3-PDO production from crude glycerol.IMPORTANCE The rapid development of the biodiesel industry has led to tremendous crude glycerol generation. Due to the presence of complex impurities, crude glycerol has low value for industry without costly purification. Obtaining novel microorganisms capable of direct and efficient bioconversion of crude glycerol to value-added products has great economic potential for industrial application. In this work, we characterized a newly isolated strain, Klebsiella pneumoniae 2e, with the capacity to efficiently produce 1,3-propanediol (1,3-PDO) from crude glycerol and demonstrated its adaptation to crude glycerol. Our work provides insights into the molecular mechanisms of K. pneumoniae 2e adaptation to crude glycerol and the expression patterns of its genes involved in 1,3-PDO biosynthesis, which will contribute to the development of industrial 1,3-PDO production from crude glycerol.

Dataset Information

High-yield production of 1,3-propanediol from glycerol by metabolically engineered Klebsiella pneumoniae.

Publications

High-yield production of 1,3-propanediol from glycerol by metabolically engineered <i>Klebsiella pneumoniae</i>.

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