Mixomics analysis of Bacillus subtilis: effect of oxygen availability on riboflavin production.

BACKGROUND: Riboflavin, an intermediate of primary metabolism, is one kind of important food additive with high economic value. The microbial cell factory Bacillus subtilis has already been proven to possess significant importance for the food industry and have become one of the most widely used riboflavin-producing strains. In the practical fermentation processes, a sharp decrease in riboflavin production is encountered along with a decrease in the dissolved oxygen (DO) tension. Influence of this oxygen availability on riboflavin biosynthesis through carbon central metabolic pathways in B. subtilis is unknown so far. Therefore the unveiled effective metabolic pathways were still an unaccomplished task till present research work. RESULTS: In this paper, the microscopic regulation mechanisms of B. subtilis grown under different dissolved oxygen tensions were studied by integrating 13C metabolic flux analysis, metabolomics and transcriptomics. It was revealed that the glucose metabolic flux through pentose phosphate (PP) pathway was lower as being confirmed by smaller pool sizes of metabolites in PP pathway and lower expression amount of ykgB at transcriptional level. The latter encodes 6-phosphogluconolactonase (6-PGL) under low DO tension. In response to low DO tension in broth, the glucose metabolic flux through Embden-Meyerhof-Parnas (EMP) pathway was higher and the gene, alsS, encoding for acetolactate synthase was significantly activated that may result due to lower ATP concentration and higher NADH/NAD+ ratio. Moreover, ResE, a membrane-anchored protein that is capable of oxygen regulated phosphorylase activity, and ResD, a regulatory protein that can be phosphorylated and dephosphorylated by ResE, were considered as DO tension sensor and transcriptional regulator respectively. CONCLUSIONS: This study shows that integration of transcriptomics, 13C metabolic flux analysis and metabolomics analysis provides a comprehensive understanding of biosynthesized riboflavin's regulatory mechanisms in B. subtilis grown under different dissolved oxygen tension conditions. The two-component system, ResD-ResE, was considered as the signal receiver of DO tension and gene regulator that led to differences between biomass and riboflavin production after triggering the shifts in gene expression, metabolic flux distributions and metabolite pool sizes.

Optimized biosynthesis of xanthan via effective valorization of orange peels using response surface methodology: A kinetic model approach.

Herein, an enhanced green production of xanthan gum has been achieved by utilizing orange peels. Response surface methodology and kinetic modeling were adapted for the process optimization and its influence on scale up production respectively. Optimal conditions for the maximum xanthan production were 1.62% acid hydrolysis, 85% carbon source of orange peel hydrolysate and 30.4°C temperature. Furthermore, the optimized treatment was conducted in the batch culture fermentor to observe the associated variations during scale up process. In bio-fermentor, to the first time ever, xanthan production along with reducing sugar conversion and utilization rates reached 30.19g/L, 69.29% and 99.99%, respectively. Employed characterization techniques of FTIR, XRD and HPLC confirmed the fermented product as xanthan gum and obtained an average molecular weight of 1.01×106g/mol. This work on account of optimized process parameters presented maximum xanthan production from a waste material.