One-step production of functional branched oligoglucosides with coupled fermentation of Pichia pastoris GS115 and Sclerotium rolfsii WSH-G01.

Endo-β-1,3-glucanase with high specific activity is a prerequisite for enzymatic preparation of valuable β-oligoglucosides. Heterologous expression in Pichia pastoris GS115 with error-prone PCR technology was implemented, and the mutant strain 7 N12 was obtained. The mutant endo-β-1,3-glucanase showed efficient specific activities for degrading curdlan (366 U mg) and scleroglucan (274.5 U mg). Thereafter, one-step production of functional branched oligoglucosides was established with coupled fermentation of Pichia pastoris and Sclerotium rolfsii. During the fermentation process, the endo-β-1,3-glucanase secreted by Pichia pastoris GS115 can efficiently hydrolyse scleroglucan metabolized by Sclerotium rolfsii WSH-G01. The maximum yields of β-oligoglucosides in the shake flasks and 7-L bioreactor reached 1.73 g L and 12.71 g L, respectively, with polymerization degrees of 2-17. The successful implementation of heterologous expression with error-prone PCR and the coupled fermentation simplified the multi-step enzymatic β-oligoglucoside preparation procedures, which makes it a potential strategy for industrial production of functional oligosaccharides.