In 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 and , but menaquinone biosynthesis was found to be incomplete in Lactobacillaceae (except ). has only two genes (, ) encoding enzymes in the biosynthetic pathway (out of eight), and has only four (, , , and ). We constructed knock-out strains of defective in , , , and (encoding the last steps in the pathway) and complemented these by expression of the extant genes from and to verify their functionality. Three of the biosynthesis genes, , , and , as well as and from , reconstituted menaquinone production and respiratory growth in the deficient strains when supplemented with heme. We then reconstituted the incomplete menaquinone biosynthesis pathway in by expressing six genes from homologous to the missing genes in a synthetic operon with two inducible promoters. Higher biomass formation was observed in carrying this operon, with an OD increase from 3.0 to 5.0 upon induction.

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8303990PMC
http://dx.doi.org/10.3390/microorganisms9071476DOI Listing

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