Association between organic nitrogen substrates and the optical purity of D-lactic acid during the fermentation by Sporolactobacillus terrae SBT-1.

The development of biotechnological lactic acid production has attracted attention to the potential production of an optically pure isomer of lactic acid, although the relationship between fermentation and the biosynthesis of highly optically pure D-lactic acid remains poorly understood. Sporolactobacillus terrae SBT-1 is an excellent D-lactic acid producer that depends on cultivation conditions. Herein, three enzymes responsible for synthesizing optically pure D-lactic acid, including D-lactate dehydrogenase (D-LDH; encoded by ldhDs), L-lactate dehydrogenase (L-LDH; encoded by ldhLs), and lactate racemase (Lar; encoded by larA), were quantified under different organic nitrogen sources and concentration to study the relationship between fermentation conditions and synthesis pathway of optically pure lactic acid. Different organic nitrogen sources and concentrations significantly affected the quantity and quality of D-lactic acid produced by strain SBT-1 as well as the synthetic optically pure lactic acid pathway. Yeast extract is a preferred organic nitrogen source for achieving high catalytic efficiency of D-lactate dehydrogenase and increasing the transcription level of ldhA2, indicating that this enzyme plays a major role in D-lactic acid formation in S. terrae SBT-1. Furthermore, lactate racemization activity could be regulated by the presence of D-lactic acid. The results of this study suggest that specific nutrient requirements are necessary to achieve a stable and highly productive fermentation process for the D-lactic acid of an individual strain.

Mathematical Modeling for Evaluating Inherent Parameters Affecting UVC Decontamination of Indicator Bacteria.

UV light is a tool associated with the denaturation of cellular components, DNA damage, and cell disruption. UV treatment is widely used in the decontamination process; however, predicting a sufficient UV dose by using traditional methods is doubtful. In this study, an in-house UVC apparatus was designed to investigate the process of the inactivation of five indicator bacteria when the initial cell concentrations and irradiation intensities varied. Both linear and nonlinear mathematical models were applied to predict the inactivation kinetics. In comparison with the Weibull and modified Chick-Watson models, the Chick-Watson model provided a good fit of the experimental data for five bacteria, Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Streptococcus faecalis, and Bacillus subtilis. The specific death rate (kd) significantly increased when the irradiation intensity (I) increased from 1.41 W/m2 to 3.02 W/m2 and 4.83 W/m2 (P < 0.05). Statistical analysis revealed no significant difference in the kd values among the groups of tested Gram-positive bacteria, Gram-negative bacteria, and B. subtilis spores, but the kd values differed among groups (P < 0.05). The death rate coefficient (k) varied from species to species. The k values of the tested Gram-positive bacteria were higher than those of the Gram-negative bacteria. The thick peptidoglycan layer in the Gram-positive membrane was responsible for UVC resistance. The high guanine-cytosine (GC) content in bacteria also contributed to UV resistance due to the less photoreactive sites on the nucleotides. This investigation provides a good understanding of bacterial inactivation induced by UVC treatment. IMPORTANCE Prevention and control measures for microbial pathogens have attracted worldwide attention due to the recent coronavirus disease 2019 pandemic. UV treatments are used as a commercial control to prevent microbial contamination in diverse applications. Microorganisms exhibit different UV sensitivities, which are often measured by the UV doses required for decreasing the number of microbial contaminants in the logarithmic order. The maximum efficacy of UV is usually observed at 254 nm (residing in the UVC range of the light spectrum). UV technology is a nonthermal physical decontamination measure that does not require any chemicals and consumes low levels of energy while leaving insignificant amounts of chemical residues or toxic compounds. Therefore, obtaining the microbial death kinetics and their intrinsic parameters provided in this study together with the UV photoreaction rate enables advancement in the design of UV treatment systems.

Draft genome sequencing of Sporolactobacillus terrae SBT-1, an efficient bacterium to ferment concentrated sugar to D-lactic acid.

Recently, the industrial-scale development of microbial D-lactic acid production has been discussed. In this study, the efficiency of the new isolate Sporolactobacillus terrae SBT-1 for producing D-lactic acid under challenge conditions was investigated. The isolate SBT-1 exhibited superior activity in fermenting a very high glucose or sucrose concentration to D-lactic acid compared to the other S. terrae isolates previously reported in the literature; therefore, SBT-1 could overcome the limitations of effective lactic acid production. In batch cultivation using 360 g/L glucose, SBT-1 produced 290.30 g/L D-lactate with a sufficiently high glucose conversion yield of 86%, volumetric productivity of 3.02 g/L h, and optical purity of 96.80% enantiomer excess. SBT-1 could also effectively utilize 440 g/L sucrose as a sole carbon source to produce 276.50 g/L lactic acid with a conversion yield of 90%, a production rate of 2.88 g/L h, and an optical purity of 98%. D-Lactic acid fermentation by two other related producers, S. inulinus NRIC1133T and S. terrae NRIC0357T, was compared with fermentation by isolate SBT-1. The experimental data revealed that SBT-1 possessed the ability to ferment relatively high glucose or sucrose concentrations to D-lactic acid without obvious catabolite repression and byproduct formation compared to the two reference strains. In draft genome sequencing of S. terrae SBT-1, the results provided here can promote further study to overcome the current limitations for the industrial-scale production of D-lactic acid.

Terrilactibacillus tamarindi sp. nov., isolated from bark of Tamarindus indica.

A Gram-stain-positive, catalase-positive, facultatively anaerobic, terminal-spore-forming rod, designated strain BCM23-1T, was isolated from bark of Tamarindus indica collected from Chiang Mai Province, Thailand. This strain produced d-lactic acid from glucose. It grew at 20-45 °C (optimum, 30 °C), pH 3.5-9 (optimum, pH 7.0) and in the presence of 1-4 % (w/v) NaCl. The cell-wall peptidoglycan contained meso-diaminopimelic acid (A1γ). The major isoprenoid quinone was menaquinone 7 (MK-7). Polar lipids analysis revealed the presence of diphosphatidylglycerol, phosphatidylglycerol, an unidentified aminophospholipid, an unidentified phospholipid and an unidentified lipid. The predominant cellular fatty acids were anteiso-C17 : 0, anteiso-C15:0, and iso-C16 : 0 when cultivated on GYP agar plates. The 16S rRNA gene sequence similarity between strain BCM23-1T and Terrilactibacillus laevilacticus NK26-11T was 98.3 %. The draft genome of BCM23-1T was 3.24 Mb in size and contained 3088 coding sequences with an in silico DNA G+C content of 37.1 mol%. The values of ANIb, ANIm and digital DNA-DNA hybridization between strain BCM23-1T and T. laevilacticus NK26-11T were 89.9, 90.8 and 40.4 %, respectively. The results of phenotypic and chemotaxonomic, 16S rRNA gene sequence similarity, and whole genome analyses support strain BCM23-1T as representing a novel species of Terrilactibacillus for which the name Terrilactibacillus tamarindi sp. nov. is proposed. The type strain is BCM23-1T (=LMG 31662T=JCM 33748T=TISTR 2841T).