New two-stage pH combined with dissolved oxygen control strategy for cyclic β-1,2 glucans synthesis.

Abstract

On the basis of a novel two-stage pH combined with dissolved oxygen (DO) control strategy in fed-batch fermentation, this research addresses the influence of pH on cyclic β-1,2-glucans (CβGs) biosynthesis and melanin accumulation during the production of CβGs by Rhizobium radiobacter ATCC 13,333. Under these optimal fermentation conditions, the maximum cell concentration and CβGs concentration in a 7-L stirred-tank fermenter were 7.94gL-1 and 3.12gL-1, which were the maximum production reported for R. radiobacter. The melanin concentration of the fermentation broth was maintained at a low level, which was beneficial to the subsequent separation and purification of the CβGs. In addition, a neutral extracellular oligosaccharide (COGs-1) purified by the two-stage pH combined with DO control strategy fermentation medium was structurally characterized. Structural analyses indicated that COGs-1 was a family of unbranched cyclic oligosaccharides composed of only β-1,2-linked D-glucopyranose residues with degree of polymerization between 17 and 23, namely CβGs. This research provides a reliable source of CβGs and structural basis for further studies of biological activity and function. KEY POINTS: • A two-stage pH combined with DO control strategy was proposed for CβGs production and melanin biosynthesis by Rhizobium radiobacter. • The final extracellular CβGs production reached 3.12 gL-1, which was the highest achieved by Rhizobium radiobacter. • The existence of CβGs could be detected by TLC quickly and accurately.