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Members of the genus Bacillus are known to play an important role in promoting plant growth and protecting plants against phytopathogenic microorganisms. In this study, 21 isolates of Bacillus spp. were obtained from the root micro-ecosystem of Suaeda glauca. Analysis of the 16S rRNA genes indicated that the isolates belong to the species Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus subtilis, Bacillus pumilus, Bacillus aryabhattai and Brevibacterium frigoritolerans. One of the interesting findings of this study is that the four strains B1, B5, B16 and B21 are dominant in rhizosphere soil. Based on gyrA, gyrB, and rpoB gene analyses, B1, B5, and B21 were identified as B. amyloliquefaciens and B16 was identified as B. velezensis. Estimation of antifungal activity showed that the isolate B1 had a significant inhibitory effect on Fusarium verticillioides, B5 and B16 on Colletotrichum capsici (syd.) Butl, and B21 on Rhizoctonia cerealis van der Hoeven. The four strains grew well in medium with 1-10% NaCl, a pH value of 5-8, and promoted the growth of Arabidopsis thaliana. Our results indicate that these strains may be promising agents for the biocontrol and promotion of plant growth and further study of the relevant bacteria will provide a useful reference for the development of microbial resources.

Biotechnological hydrogen production is considered as an environmentally sustainable alternative to petrochemical sources or electrolysis. Here, disruption of lactate dehydrogenase (LDH) and metabolic regulation via the small RNA RyhB were adopted to improve hydrogen production in the novel Klebsiella sp. strain FSoil 024. The hydrogen production of FSoil 024-L (Δldh) and FSoil 024-L/R (Δldh/RyhB) from glucose in a 5-L fermenter respectively increased by 40 and 50% compared to the wild type. When glycerol was adopted as a more favorable substrate, FSoil 024-L generated 3.3 L/L of hydrogen after 52 h of fermentation, implying its great potential for the utilization of crude glycerol to produce hydrogen. Overexpression of RyhB downregulated formate biosynthesis in FSoil 024, thereby redirecting NADH toward the hydrogen production pathway. This finding provides new insights into the role of cellular reducing power in hydrogen metabolism and establishes a rationale for improving hydrogen production.

Biotechnological production of 2,3-butanediol (2,3-BD), a versatile platform bio-chemical and a potential biofuel, is limited due to by-product toxicity. In this study, we aimed to redirect the metabolic flux toward 2,3-BD in Enterobacter aerogenes (E. aerogenes) by increasing the intracellular NADH pool. Increasing the NADH/NAD+ ratio by knocking out the NADH dehydrogenase genes (nuoC/nuoD) enhanced 2,3-BD production by up to 67% compared with wild-type E. aerogenes. When lactate dehydrogenase (ldh) was knocked out, the yield of 2,3-BD was increased by 71.2% compared to the wild type. Metabolic flux analysis revealed that upregulated expression of the sRNA RyhB led to a noteworthy shift in metabolism. The 2,3-BD titer of the best mutant Ea-2 was almost seven times higher than that of the parent strain in a 5-L fermenter. In this study, an effective metabolic engineering strategy for improved 2,3-BD production was implemented by increasing the NADH/NAD+ ratio and blocking competing pathways.

Background Biogenic 2,3-butanediol (2,3-BDO) is a high-value-added compound that can be used as a liquid fuel and a platform chemical. Bioproduction of 2,3-BDO is an environmentally friendly choice. Method and results Three recombinant derivatives of the novel Klebsiella sp. isolate FSoil 024 (WT) were constructed via different strategies including deletion of lactate dehydrogenase by λ-Red homologous recombination technology, overexpression of the small-noncoding RNA RyhB and a combination of both. The 2,3-BDO productivity of the mutants increased by 61.3%-79%, and WT-Δldh/ryhB displayed the highest 2,3-BDO yield of 42.36 mM after 24 h of shake-flask fermentation. Glucose was shown as the best carbon source for 2,3-BDO production by WT-Δldh/ryhB. In addition, higher oxygenation was favorable for ideal product synthesis. The maximal 2,3-BDO yield of WT and WT-Δldh/ryhB were increased by 23.3% and 52.5% respectively compared to the control group in the presence of 70% oxygen (V:V' = O2 :(O2 + N2 )). Conclusion and implications According to the present study, deletion of lactate dehydrogenase, RyhB overexpression, and manipulation of oxygen supply showed great impacts on cell growth, 2,3-BDO productivity and cellular metabolism of the novel isolated strain Klebsiella sp. FSoil 024. This work would also provide insights for promoting 2,3-BDO biosynthesis for industrial applications.