Scleroglucan is a high-molecular water-soluble microbial exopolysaccharide and mainly applied in the fields of petroleum, food, medicine and cosmetics. The high molecular weight of scleroglucan produced by microbial fermentation leads to low solubility, high viscosity and poor dispersibility, thus bringing a series of difficulties to extraction, preservation and application. It is important to explore suitable degradation method to adjust the molecular weight of scleroglucan for expanding its industrial application. Taking Sclerotium rolfsii WSH-G01 as a model strain, in which functional annotations of the glucanase genes were conducted by whole genome sequencing. Based on design of culture system for culture system for differential expression of β-glucanase, endogenous β-glucanase genes in S. rolfsii WSH-G01 were excavated by transcriptomics analysis. Functions of these potential hydrolases were further verified. Finally, 14 potential endogenous hydrolase genes were obtained from S. rolfsii. After heterologous overexpression in Pichia pastoris, 10 soluble enzymes were obtained and 5 of them had the activity of laminarin hydrolysis by SDS-PAGE and enzyme activity analysis. Further investigation of the 5 endogenous hydrolases on scleroglucan degradation showed that enzyme GME9860 has positive hydrolysis effect. The obtained results provide references not only for obtaining low and medium molecular weight of scleroglucan with enzymatic hydrolysis, but also for producing different molecular weight of scleroglucan during S. rolfsii fermentation process with metabolic engineering.

As a stable microbial polysaccharide, scleroglucan has extensive application in the food, medicine, and cosmetics industries. However, its large-scale industrial application is limited by its high production cost, low yield, long production time, etc. This study aims to enhance scleroglucan production by Sclerotium rolfsii WSH-G01. Based on the analysis of batch fermentation kinetics parameters, a pH-shift strategy was adopted. Through systematic kinetics analysis, a 32.4 g/L scleroglucan was accomplished. The kinetic model of the pH-shift batch fermentation process was established using a logistic equation, Luedeking-Piret equation, and a Luedeking-Piret-like equation. As decreased glucose concentration could cause decreased scleroglucan synthesis rates during the batch fermentation process, 30 g/L glucose was fed in the later phase of fermentation. As a result, scleroglucan production increased to 42 g/L, with a productivity of 0.5 g/L.h. Thus, the pH-shift strategy and feeding approach could be useful for industrial scleroglucan production.