Pleiotropic effects of lactate dehydrogenase inactivation in Lactobacillus casei

Viana, Rosa; Yebra, Marı́a J.; Galán, José Luis Márquez; Monedero, Vicente; Pérez-Martı́nez, Gaspar

doi:10.1016/j.resmic.2005.02.011

Cited by 35 publications

(33 citation statements)

References 43 publications

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“…This is in agreement with the finding that the fermentation of other hexitols (sorbitol, mannitol) was improved when the L. casei cells were grown under aeration (M. J. Yebra, unpublished observations). In addition, an L. casei mutant with impaired NADH oxidation (lactate dehydrogenase-deficient mutant) grew significantly faster when the culture was aerated (33). The degradation of one MI molecule produces two NADH equivalents in the course of the first enzymatic reactions, leading to the formation of acetyl-CoA and dihydroxyacetone phosphate (the dehydrogenase step catalyzed by IolG and the conversion of malonic semialdehyde to acetyl-CoA catalyzed by IolA).…”

Section: Discussionmentioning

confidence: 99%

Identification of a Gene Cluster EnablingLactobacillus caseiBL23 To Utilizemyo-Inositol

Yebra¹,

Zúñiga²,

Beaufils

et al. 2007

Appl Environ Microbiol

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Genome analysis of Lactobacillus casei BL23 revealed that, compared to L. casei ATCC 334, it carries a 12.8-kb DNA insertion containing genes involved in the catabolism of the cyclic polyol myo-inositol (MI). Indeed, L. casei ATCC 334 does not ferment MI, whereas strain BL23 is able to utilize this carbon source. The inserted DNA consists of an iolR gene encoding a DeoR family transcriptional repressor and a divergently transcribed iolTABCDG1G2EJK operon, encoding a complete MI catabolic pathway, in which the iolK gene probably codes for a malonate semialdehyde decarboxylase. The presence of iolK suggests that L. casei has two alternative pathways for the metabolism of malonic semialdehyde: (i) the classical MI catabolic pathway in which IolA (malonate semialdehyde dehydrogenase) catalyzes the formation of acetyl-coenzyme A from malonic semialdehyde and (ii) the conversion of malonic semialdehyde to acetaldehyde catalyzed by the product of iolK. The function of the iol genes was verified by the disruption of iolA, iolT, and iolD, which provided MI-negative strains. By contrast, the disruption of iolK resulted in a strain with no obvious defect in MI utilization. Transcriptional analyses conducted with different mutant strains showed that the iolTABCDG1G2EJK cluster is regulated by substrate-specific induction mediated by the inactivation of the transcriptional repressor IolR and by carbon catabolite repression mediated by the catabolite control protein A (CcpA). This is the first example of an operon for MI utilization in lactic acid bacteria and illustrates the versatility of carbohydrate utilization in L. casei BL23.myo-Inositol (MI) is the most abundant stereoisomer of inositol (1,2,3,4,5,6-cyclohexanehexol). It is common in soils, and it is a constituent of phytic acid (inositol hexaphosphate), which in the form of various salts (phytates) provides a major phosphate storage molecule in plant seeds. Several microorganisms, mostly inhabitants of soil, can utilize MI as a carbon source. For bacteria, the MI catabolic pathway has been elucidated for Enterobacter aerogenes (formerly Aerobacter aerogenes) (3). MI is important for the establishment of symbiosis in legume nodules. The metabolism of MI in Sinorhizobium fredii and Rhizobium leguminosarum is required for efficient nitrogen fixation and nodulation of soybeans (9, 15). MI dehydrogenase genes (e.g., iolG) and inosose dehydratase genes (iolE) have been characterized in S. fredii and Sinorhizobium meliloti (10, 15, 36), and iolA and iolDEB genes have been studied in R. leguminosarum (9). An MI utilization gene cluster has also been described for Clostridium perfringens, which was induced by MI via the IolR regulator (16), and recently, transcriptome analysis permitted the identification of an iol cluster which allowed the rapid growth of Corynebacterium glutamicum on MI (19). However, the first MI catabolic gene cluster was characterized in Bacillus subtilis, and detailed molecular data are available only for this bacterium. The B. subtilis iolRS and iolABC...

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Section: Discussionmentioning

confidence: 99%

Identification of a Gene Cluster EnablingLactobacillus caseiBL23 To Utilizemyo-Inositol

Yebra¹,

Zúñiga²,

Beaufils

et al. 2007

Appl Environ Microbiol

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“…L. casei possesses five lactate dehydrogenase genes, four of which were mutated. LSEI_2549 (ldh1) is the main L-lactate dehydrogenase and is the only one whose disruption led to a significant (25%) decrease in lactate production during growth in vitro (33) and to the synthesis of unusual end-fermentation products such as mannitol, acetoin, and ethanol (34). Thus, a complete and less efficient reorganization of sugar metabolism should strongly impact the adaptation of L. casei in the gut lumen, where the concentration and variety of sugars is limited.…”

Section: Significancementioning

confidence: 99%

Functional genomics of Lactobacillus casei establishment in the gut

Licandro

Scornec

Pédron

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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Although the composition of the gut microbiota and its symbiotic contribution to key host physiological functions are well established, little is known as yet about the bacterial factors that account for this symbiosis. We selected Lactobacillus casei as a model microorganism to proceed to genomewide identification of the functions required for a symbiont to establish colonization in the gut. As a result of our recent development of a transposonmutagenesis tool that overcomes the barrier that had prevented L. casei random mutagenesis, we developed a signature-tagged mutagenesis approach combining whole-genome reverse genetics using a set of tagged transposons and in vivo screening using the rabbit ligated ileal loop model. After sequencing transposon insertion sites in 9,250 random mutants, we assembled a library of 1,110 independent mutants, all disrupted in a different gene, that provides a representative view of the L. casei genome. By determining the relative quantity of each of the 1,110 mutants before and after the in vivo challenge, we identified a core of 47 L. casei genes necessary for its establishment in the gut. They are involved in housekeeping functions, metabolism (sugar, amino acids), cell wall biogenesis, and adaptation to environment. Hence we provide what is, to our knowledge, the first global functional genomics analysis of L. casei symbiosis.commensalism | Lactic acid bacteria T he pioneering studies that led to the characterization of the gut microbiota were reviewed in 2001 (1). These studies and recent investigations have revealed mutualistic functions (2), including a barrier effect against allogenic microbes (3), fermentation of complex sugars (4, 5), and maturation and homeostasis of the immune system (6). Recent metagenomic studies have revealed an extraordinary diversity of genes constituting the gut microbiome (7), opening the way to correlative studies linking microbiome diversity, homeostasis, and diseases (5,8,9).In parallel, some representative species, i.e., "model symbionts," now are being studied functionally (10). As it was done for pathogens, it is essential to develop the cellular microbiology of symbionts and particularly to identify the genes required for their establishment and persistence in the gut. Transcriptomic profiling identified up-regulated genes linked to metabolic functions, stress responses, and pili synthesis during early colonization (11-13). Comparative genomics among Lactobacilli identified strain-specific candidate genes for extended colonization: In Lactobacillus rhamnosus, persistence was attributed to an spaCBA locus encoding LPXTG-like pilins (14), and in Lactobacillus johnsonii it was attributed to specific glycosyltransferases, a phosphotransfer system, and a protease (15). Otherwise, a functional in vivo screening based on the expression of a genomic library of Bacteroides fragilis identified a locus encoding polysaccharide utilization as essential for stable colonization of murine colonic crypts (16). Alternatively, colonization of germ-free mic...

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“…The bacterial genera that form lactic acid are often called lactic acid bacteria (LAB). The fermentative metabolism of LAB is characterized by the glycolytic breakdown of carbohydrates, and a late step in this pathway is distinguished by the conversion of pyruvate into lactic acid, a reaction that oxidizes the NADH formed during glycolysis, thus maintaining cellular redox balance (7). The enzymes responsible for the conversion of pyruvate to lactic acid and back are lactate dehydrogenases (LDHs), which fall into two broad classes: NAD-dependent lactate dehydrogenases (nLDHs) and NAD-independent lactate dehydrogenases (iLDHs).…”

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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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Pleiotropic effects of lactate dehydrogenase inactivation in Lactobacillus casei

Cited by 35 publications

References 43 publications

Identification of a Gene Cluster EnablingLactobacillus caseiBL23 To Utilizemyo-Inositol

Identification of a Gene Cluster EnablingLactobacillus caseiBL23 To Utilizemyo-Inositol

Functional genomics of Lactobacillus casei establishment in the gut

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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