Major role of NAD-dependent lactate dehydrogenases in aerobic lactate utilization in Lactobacillus plantarum during early stationary phase - PubMed (original) (raw)

Major role of NAD-dependent lactate dehydrogenases in aerobic lactate utilization in Lactobacillus plantarum during early stationary phase

Philippe Goffin et al. J Bacteriol. 2004 Oct.

Abstract

NAD-independent lactate dehydrogenases are commonly thought to be responsible for lactate utilization during the stationary phase of aerobic growth in Lactobacillus plantarum. To substantiate this view, we constructed single and double knockout mutants for the corresponding genes, loxD and loxL. Lactate-to-acetate conversion was not impaired in these strains, while it was completely blocked in mutants deficient in NAD-dependent lactate dehydrogenase activities, encoded by the ldhD and ldhL genes. We conclude that NAD-dependent but not NAD-independent lactate dehydrogenases are involved in this process.

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Figures

FIG. 1.

FIG. 1.

(A) Metabolic pathways of lactic acid production and utilization in L. plantarum in aerobic conditions as proposed in the literature. (B) Proposed pathway for

d

- and

l

-lactate utilization in the stationary phase of aerobic growth of L. plantarum. [1], glycolysis; [2],

l

-nLDH (LdhL); [3],

d

-nLDH (LdhD); [4],

l

-iLDH; [5],

d

-iLDH; [6], pyruvate oxidase; [7], acetate kinase; [8], lactate racemase; [9], NADH oxidase; [10], NADH peroxidase; X, unknown electron acceptor.

FIG. 2.

FIG. 2.

l

-iLDH (white bars) and

d

-iLDH (black bars) activities in L. plantarum NCIMB8826 (wild type), TF101 (ΔldhL), TF102 (ldhD::cat), PG901 (ΔloxL), PG801 (loxD::cat), and PG701 (ΔloxL loxD::cat). One unit corresponds to 1 μmol of DCPIP reduced min−1 mg of total protein−1. Total protein concentration was measured using the Bradford method (1). These data represent average values (from at least three individual measurements) with their standard deviations.

FIG. 3.

FIG. 3.

Growth curves of L. plantarum NCIMB8826 (A), TF101 (B), and TF102 (C) in MRS-CA with 0.2% (wt/vol) glucose and shaking at 28°C. The growth was measured by monitoring the OD (600 nm). Concentrations of

d

-lactate,

l

-lactate, acetate, and glucose were determined in the supernatant. Concentrations of lactate and acetate are given as the difference between the measured concentration and the initial concentration in the culture medium.

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References

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