Draft Genome Sequence of Sporolactobacillus inulinus Strain CASD, an Efficient D-Lactic Acid-Producing Bacterium with High-Concentration Lactate Tolerance Capability

Abstract

Sporolactobacillus inulinus CASD is an efficient D-lactic acid producer with high optical purity. Here we report for the first time the draft genome sequence of S. inulinus (2,930,096 bp). The large number of annotated two-component system genes makes it possible to explore the mechanism of extraordinary lactate tolerance of S. inulinus CASD.Lactic acid, the most important hydroxycarboxylic acid, is regarded as a feedstock for the green chemistry of the future. Besides its versatile applications in the food, pharmaceutical, textile, and leather industries, lactic acid is also a building block for biodegradable plastic, mainly poly(L-lactic acid), which has gained substantial attention in the new wave of industrial biotechnology (7, 4). The development of a stereocomplex of poly(L-lactic acid) and poly(D-lactic acid) is considered an advance in poly(lactic acid) modification, in which the melting point of this stereocomplex could reach the same temperature as that of other petroleum-based polymers (3). Therefore, the potential utilization of D-lactic acid has attracted increased attention to its production by using sustainable bioresource materials (8,12).Sporolactobacillus is a genus of anaerobic endosporeforming, Gram-positive, motile, rod-shaped lactic acid bacteria. Sporolactobacillus inulinus, the type species of the genus, was originally named Sporolactobacillus (subgen.

The D-Lactate Dehydrogenase from Sporolactobacillus inulinus Also Possessing Reversible Deamination Activity

“…S . inulinus CASD was used as the source for cloning DLDH744 gene [ 13 ]. Escherichia coli strains were grown in Luria-Bertani (LB) medium aerobically and set on a rotary shaker.…”

NADP + -Preferring d -Lactate Dehydrogenase from Sporolactobacillus inulinus

“…The draft genome sequence of S. inulinus CASD has been deposited in GenBank under accession number AFVQ00000000 (14). According to the nucleotide sequence of the annotated gene (NODE_744_orf00003, 1,008 bp including the stop codon), a forward primer (5=-GGTCATGAAAATCATTATGTTCAG-3=) and a reverse primer (5=-GCAAGCTTGTTTTCATCAGCTACTTTGTT-3=) were designed to introduce BspHI and HindIII restriction sites on either side of the gene.…”

“…Based on the annotated genome sequence (14), the gene product was designated D-lactate dehydrogenase 744 (DLDH744). Interestingly, we found that DLDH744 preferentially uses NADPH as a coenzyme, by contrasting the apparent K m values for NADH and NADPH.…”

Draft Genome Sequence of Sporolactobacillus inulinus NBRC 111894, Isolated from Kôso, a Japanese Sugar-Vegetable Fermented Beverage

“…The VISTA servers (http://genome.lbl.gov/cgi-bin/WGVistaInput) (8, 9), with their default settings, were used for gene comparisons, with the Sporolactobacillus inulinus CASD (10) genome being applied as a reference. S. inulinus NBRC 111894 showed three annotated genes relating to lipopolysaccharide biosynthesis and metabolism; in contrast, S. inulinus CASD showed only one.…”