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

Abstract

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 13 C 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.

Bacillus subtilis with a 1% population enhances the activity of Enterococcus faecalis with a 99% population

“…B. subtilis has been widely used to produce amino acids ( 17 ) and vitamins ( 18 ). Since oxygen is required for the production of these compounds ( 19, 20 ), B. subtilis probably produced such reducing agents under oxic conditions, resulting in the lower ORP values observed in this study.…”

Improving riboflavin production by modifying related metabolic pathways in Ba cillus subtilis

“…2012) Under oxygen‐limited conditions, the metabolic flux of the PP pathway decreases, and the transcription level of ykgB decreases (Hu et al . 2017). Overexpression of ykgB has a certain effect on the synthesis of the downstream metabolite uridine of PRPP (Zhang and Wang 2020).…”

“…Some studies have shown that under hypoxic conditions, the metabolic flux of the PP pathway decreases, accompanied by a decrease in ykgB transcription levels (Hu et al . 2017). In B. subtilis , there is a balanced relationship between de novo IMP and the UMP synthesis pathway (Liu et al .…”

Biosurfactant-Mediated Membrane Depolarization Maintains Viability during Oxygen Depletion in Bacillus subtilis

“…The increase in growth yield due to surfactin suggests that oxygen can be limiting even in aerobic cultures and that increasing oxygen availability increases yield. This finding has industrial implications as B. subtilis is commonly used to produce many enzymes and antibiotics [53], and maintaining oxygen levels is critical to optimize production of the desired compounds [54,55]. In addition to oxygen, surfactin might also facilitate growth by increasing nutrient diffusion during late exponential phase when nutrients become limiting.…”