Heterologous production of L-pipecolic acid in Corynebacterium glutamicum

Wagner, Nina; Steinkämper, Anne; Biener, Richard; Schwartz, Dirk

doi:10.1016/j.jbiotec.2010.09.523

Cited by 2 publications

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“…L‐PA can be obtained by chemical synthesis or by biocatalysis using L‐lysine cyclodeaminase . However, the chemical process shows low conversion yields and the biocatalytic process requires very high enzyme loading due to the low turnover numbers (around 0.6 s −1 ) of L‐lysine cyclodeaminases .…”

Section: Introductionmentioning

confidence: 99%

Fermentative production of L‐pipecolic acid from glucose and alternative carbon sources

Pérez-García

Risse

Friehs

et al. 2017

Biotechnology Journal

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Corynebacterium glutamicum is used for the million-ton scale production of amino acids and has recently been engineered for production of the cyclic non-proteinogenic amino acid L-pipecolic acid (L-PA). In this synthetic pathway L-lysine was converted to L-PA by oxidative deamination, dehydration and reduction by L-lysine 6-dehydrogenase (deaminating) from Silicibacter pomeroyi and pyrroline 5-carboxylate reductase from C. glutamicum. However, production of L-PA occurred as by-product of L-lysine production only. Here, the author show that abolishing L-lysine export by the respective gene deletion resulted in production of L-PA as major product without concomitant lysine production while the specific growth rate was reduced due to accumulation of high intracellular lysine concentrations. Increasing expression of the genes encoding L-lysine 6-dehydrogenase and pyrroline 5-carboxylate reductase in C. glutamicum strain PIPE4 increased the L-PA titer to 3.9 g L , and allowed faster growth and, thus, a higher volumetric productivity of 0.08 ± 0.00 g L h respectively. Secondly, expression of heterologous genes for utilization of glycerol, xylose, glucosamine, and starch in strain PIPE4 enabled L-PA production from these alternative carbon sources. Third, in a glucose/sucrose-based fed-batch fermentation with C. glutamicum PIPE4 L-PA was produced to a titer of 14.4 g L with a volumetric productivity of 0.21 g L h and an overall yield of 0.20 g g .

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Section: Introductionmentioning

confidence: 99%

Fermentative production of L‐pipecolic acid from glucose and alternative carbon sources

Pérez-García

Risse

Friehs

et al. 2017

Biotechnology Journal

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“…Biotransformation of racemic DL-lysine with E. coli overproducing L-lysine α-oxidase from S. japonicus yielded high titers (about 45 g L −1 ) of optically pure L-PA (Tani et al 2015). The functionality of L-lysine cyclodeaminase from Actinoplanes friuliensis has been shown in C. glutamicum (Wagner et al 2010), and the enzyme from Streptomyces. pristinaespiralis has been used in biotransformations of L-lysine to L-PA with recombinant E. coli (Tsotsou and Barbirato 2007).…”

Section: Discussionmentioning

confidence: 98%

Engineering Corynebacterium glutamicum for fast production of l-lysine and l-pipecolic acid

Pérez-García

Peters‐Wendisch

Wendisch

2016

Appl Microbiol Biotechnol

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The Gram-positive Corynebacterium glutamicum is widely used for fermentative production of amino acids. The world production of L-lysine has surpassed 2 million tons per year. Glucose uptake and phosphorylation by C. glutamicum mainly occur by the phosphotransferase system (PTS) and to lesser extent by inositol permeases and glucokinases. Heterologous expression of the genes for the high-affinity glucose permease from Streptomyces coelicolor and Bacillus subtilis glucokinase fully compensated for the absence of the PTS in Δhpr strains. Growth of PTS-positive strains with glucose was accelerated when the endogenous inositol permease IolT2 and glucokinase from B. subtilis were overproduced with balanced translation initiation rates using plasmid pEKEx3-IolTBest. When the genome-reduced C. glutamicum strain GRLys1 carrying additional in-frame deletions of sugR and ldhA to derepress glycolytic and PTS genes and to circumvent formation of L-lactate as by-product was transformed with this plasmid or with pVWEx1-IolTBest, 18 to 20 % higher volumetric productivities and 70 to 72 % higher specific productivities as compared to the parental strain resulted. The non-proteinogenic amino acid L-pipecolic acid (L-PA), a precursor of immunosuppressants, peptide antibiotics, or piperidine alkaloids, can be derived from L-lysine. To enable production of L-PA by the constructed L-lysine-producing strain, the L-lysine 6-dehydrogenase gene lysDH from Silicibacter pomeroyi and the endogenous pyrroline 5-carboxylate reductase gene proC were overexpressed as synthetic operon. This enabled C. glutamicum to produce L-PA with a yield of 0.09 ± 0.01 g g(-1) and a volumetric productivity of 0.04 ± 0.01 g L(-1) h(-1).To the best of our knowledge, this is the first fermentative process for the production of L-PA from glucose.

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Heterologous production of L-pipecolic acid in Corynebacterium glutamicum

Cited by 2 publications

References 0 publications

Fermentative production of L‐pipecolic acid from glucose and alternative carbon sources

Fermentative production of L‐pipecolic acid from glucose and alternative carbon sources

Engineering Corynebacterium glutamicum for fast production of l-lysine and l-pipecolic acid

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