Dataset Information

Analysis and Reconstitution of the Menaquinone Biosynthesis Pathway in Lactiplantibacillus plantarum and Lentilactibacillus buchneri.

ABSTRACT: In Lactococcus lactis and some other lactic acid bacteria, respiratory metabolism has been reported upon supplementation with only heme, leading to enhanced biomass formation, reduced acidification, resistance to oxygen, and improved long-term storage. Genes encoding a complete respiratory chain with all components were found in genomes of L. lactis and Leuconostoc mesenteroides, but menaquinone biosynthesis was found to be incomplete in Lactobacillaceae (except L. mesenteroides). Lactiplantibacillus plantarum has only two genes (menA, menG) encoding enzymes in the biosynthetic pathway (out of eight), and Lentilactobacillus buchneri has only four (menA, menB, menE, and menG). We constructed knock-out strains of L. lactis defective in menA, menB, menE, and menG (encoding the last steps in the pathway) and complemented these by expression of the extant genes from Lactipl. plantarum and Lent. buchneri to verify their functionality. Three of the Lactipl. plantarum biosynthesis genes, lpmenA1, lpmenG1, and lpmenG2, as well as lbmenB and lbmenG from Lent. buchneri, reconstituted menaquinone production and respiratory growth in the deficient L. lactis strains when supplemented with heme. We then reconstituted the incomplete menaquinone biosynthesis pathway in Lactipl. plantarum by expressing six genes from L. lactis homologous to the missing genes in a synthetic operon with two inducible promoters. Higher biomass formation was observed in Lactipl. plantarum carrying this operon, with an OD₆₀₀ increase from 3.0 to 5.0 upon induction.

SUBMITTER: Watthanasakphuban N

PROVIDER: S-EPMC8303990 | biostudies-literature | 2021 Jul

REPOSITORIES: biostudies-literature

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Analysis and Reconstitution of the Menaquinone Biosynthesis Pathway in Lactiplantibacillus plantarum and Lentilactibacillus buchneri.

Watthanasakphuban Nisit N Virginia Ludovika Jessica LJ Haltrich Dietmar D Peterbauer Clemens C

Microorganisms 20210709 7

In Lactococcus lactis and some other lactic acid bacteria, respiratory metabolism has been reported upon supplementation with only heme, leading to enhanced biomass formation, reduced acidification, resistance to oxygen, and improved long-term storage. Genes encoding a complete respiratory chain with all components were found in genomes of L. lactis and Leuconostoc mesenteroides, but menaquinone biosynthesis was found to be incomplete in Lactobacillaceae (except L. mesent ...[more]

PMID: 34361912

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Project description:This study aimed to investigate the effects of Lactiplantibacillus plantarum (L. plantarum) or Lentilactobacillus buchneri (L. buchneri) on the fermentation characteristics and microbial community of oat silages in response to additives. The oat was harvested at the milk stage of maturity and was chopped into 30 mm in size. Then, the oat was treated with distilled water (control, CON treatment), L. buchneri, and L. plantarum; the addition of L. buchneri and L. plantarum was in 1 × 106 colony-forming units/g of fresh matter and stored at room temperature (25°C). Results showed that the addition of L. plantarum inoculations could increase the lactic acid concentrations of oat silages compared with the control, and the addition of L. buchneri could increase the acetic acid concentrations, whereas the addition of L. plantarum could decrease the fiber contents and increase the crude protein content. The Shannon index of bacterial community was markedly (P < 0.05) lower in the L. buchneri- and L. plantarum-treated oat silages, and the Shannon index of fungal community was significantly (P < 0.05) higher in L. buchneri- and L. plantarum-treated oat silages compared with the CON treatment for 7 and 10 days of ensiling. From 7 to 90 days of ensiling, Lactobacillus was the dominant genus during the whole fermentation process in the three treatments. The homofermentative L. plantarum regulated the fermentation quality and microbial community by enhancing the Emden-Meyerhoff pathway, phosphoketolase pathway, and pentosephosphate pathway, and the heterofermentative L. buchneri modulated the ensiling performance and microbial community via improving the pentosephosphate pathway. These results suggested that the addition of lactic acid bacteria could improve the ensiling performance by regulating the microbial community in oat silage, and L. plantarum was more beneficial than L. buchneri for enhancing the fermentation quality.IMPORTANCEEnsiled whole-plant oats are an important feedstuff for ruminants in large parts of the world. Oat silage is rich in dietary fibers, minerals, vitamins, and phytochemicals beneficial to animal health. The fermentation of oat silage is a complex biochemical process that includes interactions between various microorganisms. The activity of many microbes in silage may cause an extensive breakdown of nutrition and lead to undesirable fermentation. Moreover, it is difficult to make high-quality oat silage because the number of epiphytic lactic acid bacterium microflora was lower than the requirement. Understanding the complex microbial community during the fermentation process and its relationship with community functions is therefore important in the context of developing improved fermentation biotechnology systems. These results suggested that the addition of Lactobacillus plantarum or Lactobacillus buchneri regulated the ensiling performance and microbial community in oat silage by shaping the metabolic pathways.

Project description:CLA (conjugated linoleic acid) has attracted substantial attention due to its physiological functions, including regulating immunity, reducing obesity, and contributing to cancer suppression. In Lactiplantibacillus plantarum, CLA oleate hydratase (CLA-HY), CLA short-chain dehydrogenase (CLA-DH), and CLA acetoacetate decarboxylase (CLA-DC) catalyze the biotransformation of linoleic acid (LA) to CLA. However, the underlying transcriptional regulation mechanism of this pathway remains largely unknown. In this study, the potential transcriptional regulators that might bind to the cla promoter of L. plantarum AR195 were investigated by DNA pulldown. Interestingly, ArgR2, the transcriptional regulator of arginine metabolism, was identified as a potential regulator involved in the regulation of CLA biotransformation. Electrophoretic mobility shift assay (EMSA) and molecular interaction results demonstrated the specific binding of ArgR2 to the regulatory region of the cla operon. The knockout of argR2 led to the downregulation of cla-dh and cla-dc by 91% and 34%, respectively, resulting in a decline in the CLA yield by 14%. A segmental EMSA revealed that ArgR2 bound to three distinct sites in the cla regulatory region, and these binding sites were highly conserved and rich in AT. The regulatory mechanism of ArgR2 on CLA biosynthesis further expanded our knowledge of the regulatory mechanism of CLA biosynthesis in L. plantarum and laid the theoretical foundation for the production and application of CLA. IMPORTANCE CLA (conjugated linoleic acid) has received extensive attention owing to its important physiological functions. CLA from natural sources is far from meeting people's demands. Lactic acid bacteria can efficiently synthesize cis-9,trans-11-CLA and trans-10,cis-12-CLA, which possess physiological activities. However, little is known about the regulatory mechanism. In this study, we identified that the biosynthesis of CLA in L. plantarum AR195 was transcriptionally regulated by the arginine biosynthesis regulatory protein ArgR2. The regulation mechanism of ArgR2 on CLA biosynthesis lays a theoretical foundation for the regulation of CLA synthesis and industrial production.

Dataset Information

Analysis and Reconstitution of the Menaquinone Biosynthesis Pathway in Lactiplantibacillus plantarum and Lentilactibacillus buchneri.

Publications

Analysis and Reconstitution of the Menaquinone Biosynthesis Pathway in <i>Lactiplantibacillus plantarum</i> and <i>Lentilactibacillus buchneri</i>.

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Dataset Information

Analysis and Reconstitution of the Menaquinone Biosynthesis Pathway in Lactiplantibacillus plantarum and Lentilactibacillus buchneri.

Publications

Analysis and Reconstitution of the Menaquinone Biosynthesis Pathway in &lt;i&gt;Lactiplantibacillus plantarum&lt;/i&gt; and &lt;i&gt;Lentilactibacillus buchneri&lt;/i&gt;.

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OmicsDI is part of the ELIXIR infrastructure

Tweets

Analysis and Reconstitution of the Menaquinone Biosynthesis Pathway in <i>Lactiplantibacillus plantarum</i> and <i>Lentilactibacillus buchneri</i>.