Medium optimization of a hydrophilic solvent-stable protease from Serratia sp. SYBC H

Abstract

Two statistical methods were used for medium optimization for a hydrophilic solvent-stable protease production by Serratia sp. SYBC H with duckweed as the nitrogen source. Orthogonal design was applied to find the significant variables, then response surface methodology (RSM), including Box–Behnken central composite experiments, was used to determine the optimal concentrations and interaction of the significant variables. Results demonstrated that duckweed powder, wheat flour, Tween 80, sodium chloride had significant effects on the solvent-stable protease production. The interaction between duckweed and wheat flour was significant. The optimal level of the variables for the maximum protease production was duckweed 43.9 g/L, wheat flour 20 g/L, sodium chloride 0.08 M, Tween 80 1% v/v, initial pH 11.0, and inoculum size 7% v/v. The maximum protease activity reached 1922.8 U/mL in the optimized medium, with about 18.3-fold higher than that in the unoptimized medium. Most importantly, the protease from Serratia sp. SYBC H has successfully catalyzed the specific acylation of sucrose in a two-solvent medium consisting of pyridine and n-hexane (1:1, v/v), and non-specific acylation of sucrose in anhydrous DMSO. These results demonstrated that the protease from Serratia sp. SYBC H is a solvent-stable protease and it could be an ideal biocatalyst for sugar esters syntheses in non-aqueous media.