Biochemical characterisation of recombinant Streptomyces pristinaespiralis l-lysine cyclodeaminase

Tsotsou, Georgia Eleni; Barbirato, Fabien

doi:10.1016/j.biochi.2006.12.008

Cited by 26 publications

(28 citation statements)

References 39 publications

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“…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). However, since L-lysine cyclodeaminases show low turnover numbers (about 0.6 s −1 ), only 4.5 mM L-PA was produced per hour using 1 g L −1 pure enzyme (Tsotsou and Barbirato 2007).…”

Section: Discussionmentioning

confidence: 99%

“…pristinaespiralis has been used in biotransformations of L-lysine to L-PA with recombinant E. coli (Tsotsou and Barbirato 2007). However, since L-lysine cyclodeaminases show low turnover numbers (about 0.6 s −1 ), only 4.5 mM L-PA was produced per hour using 1 g L −1 pure enzyme (Tsotsou and Barbirato 2007). All biotransformations with isolated enzymes or whole cells use L-or DL-lysine as substrate for conversion to L-PA, while to the best of our knowledge, a fermentative route for production of L-PA from glucose is presented here for the first time.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

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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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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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show abstract

“…At pH 5.0, the L-pipecolic acid concentration was decreased 55.8% in comparison with the pH at 7.5. Although the optimal pH of whole-cell biocatalyst was different from the optimal pH of purified enzyme [16], the whole-cell biocatalyst was obviously more stable in wider pH ranges. This might be the differences between the environment of intracellular enzyme and free purified enzyme.…”

Section: Effect Of Ph and Temperature On The Whole-cell Biocatalystmentioning

confidence: 94%

“…Recently, studies on the biosynthesis of rapamysin in Streptomyces hygroscopicus found that lysine cyclodeaminase (LCD) could directly catalyze the conversion of L-lysine to L-pipecolic acid in one step [12]. Tsotsou et al expressed LCD in Escherichia coli and investigated the biocatalytic properties of this enzyme [16], whereas Walsh et al, after heterologous expression and purification, demonstrated the mechanisms of LCD and confirmed the cofactor of this enzyme [12]. When compared with purified enzymes, whole-cell biocatalyst could simplify the process, leading to a relatively high reaction efficiency [17].…”

Section: Introductionmentioning

confidence: 99%

Enhanced conversion of L-lysine to L-pipecolic acid using a recombinant Escherichia coli containing lysine cyclodeaminase as whole-cell biocatalyst

Ying

Wang

et al. 2015

Journal of Molecular Catalysis B: Enzymatic

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“…Activity of the dihydrodipicolinate synthase (DHDPS, EC 4.2.1.52) was measured using a coupled assay originally described by Sibarani et al [36]. The activity of lysine cyclodeaminase (LCD, EC 4.3.1.28) was measured by the method of Tsotsou and Barbirato [40].…”

Section: Enzymes Activity Assaysmentioning

confidence: 99%

Comparative metabolic profiling reveals the key role of amino acids metabolism in the rapamycin overproduction by Streptomyces hygroscopicus

Wang

Liu

et al. 2015

Journal of Industrial Microbiology and Biotechnology

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Rapamycin is an important natural macrolide antibiotic with antifungal, immunosuppressive and anticancer activity produced by Streptomyces hygroscopicus. In this study, a mutant strain obtained by ultraviolet mutagenesis displayed higher rapamycin production capacity compared to the wild-type S. hygroscopicus ATCC 29253. To gain insights into the mechanism of rapamycin overproduction, comparative metabolic profiling between the wild-type and mutant strain was performed. A total of 86 metabolites were identified by gas chromatography-mass spectrometry. Pattern recognition methods, including principal component analysis, partial least squares and partial least squares discriminant analysis, were employed to determine the key biomarkers. The results showed that 22 potential biomarkers were closely associated with the increase of rapamycin production and the tremendous metabolic difference was observed between the two strains. Furthermore, metabolic pathway analysis revealed that amino acids metabolism played an important role in the synthesis of rapamycin, especially lysine, valine, tryptophan, isoleucine, glutamate, arginine and ornithine. The inadequate supply of amino acids, or namely "nitrogen starvation" occurred in the mutant strain. Subsequently, the exogenous addition of amino acids into the fermentation medium of the mutant strain confirmed the above conclusion, and rapamycin production of the mutant strain increased to 426.7 mg/L after adding lysine, approximately 5.8-fold of that in the wild-type strain. Finally, the results of real-time PCR and enzyme activity assays demonstrated that dihydrodipicolinate synthase involved with lysine metabolism played vital role in the biosynthesis of rapamycin. These findings will provide a theoretical basis for further improving production of rapamycin.

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Biochemical characterisation of recombinant Streptomyces pristinaespiralis l-lysine cyclodeaminase

Cited by 26 publications

References 39 publications

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

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

Enhanced conversion of L-lysine to L-pipecolic acid using a recombinant Escherichia coli containing lysine cyclodeaminase as whole-cell biocatalyst

Comparative metabolic profiling reveals the key role of amino acids metabolism in the rapamycin overproduction by Streptomyces hygroscopicus

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