Kinetic characterisation of recombinant Corynebacterium glutamicum NAD+-dependent LDH over-expressed in E. coli and its rescue of an lldD − phenotype in C. glutamicum: the issue of reversibility re-examined

Sharkey, Michael A.; Maher, Marcus A.; Guyonvarch, Armel; Engel, Paul C.

doi:10.1007/s00203-011-0711-z

Cited by 11 publications

(5 citation statements)

References 38 publications

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“…Disruption of l -iLDH resulted in the loss of ability to utilize l -lactate as the sole carbon source [9], [28]. Constitutively expressed NAD-dependent l -lactate dehydrogenase would enable the l -iLDH inactivation mutant of C. glutamicum to grow on l -lactate [29]. In the present study, l -iLDH from P. stutzeri SDM was purified and characterized.…”

Section: Resultsmentioning

confidence: 91%

NAD-Independent L-Lactate Dehydrogenase Is Required for L-Lactate Utilization in Pseudomonas stutzeri SDM

et al. 2012

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BackgroundVarious Pseudomonas strains can use l-lactate as their sole carbon source for growth. However, the l-lactate-utilizing enzymes in Pseudomonas have never been identified and further studied.Methodology/Principal FindingsAn NAD-independent l-lactate dehydrogenase (l-iLDH) was purified from the membrane fraction of Pseudomonas stutzeri SDM. The enzyme catalyzes the oxidation of l-lactate to pyruvate by using FMN as cofactor. After cloning its encoding gene (lldD), l-iLDH was successfully expressed, purified from a recombinant Escherichia coli strain, and characterized. An lldD mutant of P. stutzeri SDM was constructed by gene knockout technology. This mutant was unable to grow on l-lactate, but retained the ability to grow on pyruvate.Conclusions/SignificanceIt is proposed that l-iLDH plays an indispensable function in Pseudomonas l-lactate utilization by catalyzing the conversion of l-lactate into pyruvate.

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Section: Resultsmentioning

confidence: 91%

NAD-Independent L-Lactate Dehydrogenase Is Required for L-Lactate Utilization in Pseudomonas stutzeri SDM

et al. 2012

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“…). The nLDH/Etf in A. woodii and l ‐nLDHs in C. glutamicum and Lactococcus lactis were recently reported to enable these microorganisms to grow on l ‐lactate (Sharkey et al ., ; Zhao et al ., ; Schink, ; Weghoff et al ., ). In this study, we first demonstrated that a d ‐nLDH encoding gene ldhA in P. putida KT2440 had no role in its d ‐lactate utilization by mutation analysis.…”

Section: Discussionmentioning

confidence: 99%

Coexistence of two d‐lactate‐utilizing systems in Pseudomonas putida KT2440

Zhang

Jiang

Sheng

et al. 2016

Environ Microbiol Rep

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It is advantageous for rhizosphere-dwelling microorganisms to utilize organic acids such as lactate. Pseudomonas putida KT2440 is one of the most widely studied rhizosphere-dwelling model organisms. The P. putida KT2440 genome contains an NAD-dependent d-lactate dehydrogenase encoding gene, but mutation of this gene does not play a role in d-lactate utilization. Instead, it was found that d-lactate utilization in P. putida KT2440 proceeds via a multidomain NAD-independent d-lactate dehydrogenase with a C-terminal domain containing several Fe-S cluster-binding motifs (Fe-S d-iLDH) and glycolate oxidase, which is widely distributed in various microorganisms. Both Fe-S d-iLDH and glycolate oxidase were identified to be membrane-bound proteins. Neither Fe-S d-iLDH nor glycolate oxidase is constitutively expressed but both of them can be induced by either enantiomer of lactate in P. putida KT2440. This study shows a case in which an environmental microbe contains two types of enzymes specific for d-lactate utilization.

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“…nLDHs are cytoplasmic proteins that catalyze the conversion of pyruvate to lactic acid in a reversible or irreversible manner. iLDHs are membrane bound, and they catalyze the oxidation of lactic acid to pyruvate (8). Members of these two classes exist in a stereospecific (L-or D-specific) form.…”

mentioning

confidence: 99%

Major Role of NAD-Dependent Lactate Dehydrogenases in the Production of l -Lactic Acid with High Optical Purity by the Thermophile Bacillus coagulans

Wang

Cai

Zhu

et al. 2014

Appl Environ Microbiol

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Bacillus coagulans 2-6 is an excellent producer of optically pure L-lactic acid. However, little is known about the mechanism of synthesis of the highly optically pure L-lactic acid produced by this strain. Three enzymes responsible for lactic acid production-NAD-dependent L-lactate dehydrogenase (L-nLDH; encoded by ldhL), NAD-dependent D-lactate dehydrogenase (D-nLDH; encoded by ldhD), and glycolate oxidase (GOX)-were systematically investigated in order to study the relationship between these enzymes and the optical purity of lactic acid. Lactobacillus delbrueckii subsp. bulgaricus DSM 20081 (a D-lactic acid producer) and Lactobacillus plantarum subsp. plantarum DSM 20174 (a DL-lactic acid producer) were also examined in this study as comparative strains, in addition to B. coagulans. The specific activities of key enzymes for lactic acid production in the three strains were characterized in vivo and in vitro, and the levels of transcription of the ldhL, ldhD, and GOX genes during fermentation were also analyzed. The catalytic activities of L-nLDH and D-nLDH were different in L-, D-, and DL-lactic acid producers. Only L-nLDH activity was detected in B. coagulans 2-6 under native conditions, and the level of transcription of ldhL in B. coagulans 2-6 was much higher than that of ldhD or the GOX gene at all growth phases. However, for the two Lactobacillus strains used in this study, ldhD transcription levels were higher than those of ldhL. The high catalytic efficiency of L-nLDH toward pyruvate and the high transcription ratios of ldhL to ldhD and ldhL to the GOX gene provide the key explanations for the high optical purity of L-lactic acid produced by B. coagulans 2-6.

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Kinetic characterisation of recombinant Corynebacterium glutamicum NAD+-dependent LDH over-expressed in E. coli and its rescue of an lldD − phenotype in C. glutamicum: the issue of reversibility re-examined

Cited by 11 publications

References 38 publications

NAD-Independent L-Lactate Dehydrogenase Is Required for L-Lactate Utilization in Pseudomonas stutzeri SDM

NAD-Independent L-Lactate Dehydrogenase Is Required for L-Lactate Utilization in Pseudomonas stutzeri SDM

Coexistence of two d‐lactate‐utilizing systems in Pseudomonas putida KT2440

Major Role of NAD-Dependent Lactate Dehydrogenases in the Production of l -Lactic Acid with High Optical Purity by the Thermophile Bacillus coagulans

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