Improved Expression and Optimization of Trehalose Synthase by Regulation of P glv in Bacillus subtilis

Abstract

Trehalose synthase (TreS) converts maltose to trehalose, which has several important functions; therefore, enhancing TreS expression is desirable. Here, a recombinant Bacillus subtilis W800N (ΔamyE)-P glv strain was constructed to achieve enhanced expression of TreS. Process optimization strategies were developed to improve the expression level of TreS in B. subtilis W800N (ΔamyE)-P glv . Intracellular activity of TreS was induced using 60 g/L of maltose in shake flask culture. The protein activity reached 5211 ± 134 U/g at 33 °C and pH 7.0 in Luria-Bertani medium. A fed-batch fermentation strategy was applied in a 30 L fermenter containing 18 L terrific broth to achieve high cell density by replacing glycerol with high maltose syrup as a carbon source and an inducer. After 32 h of fermentation, recombinant B. subtilis W800N (ΔamyE)-P glv activity reached 6850 ± 287 U/g dry cell weight. Our results demonstrate the efficiency of the P glv promoter in increasing the expression of TreS in B. subtilis W800N (ΔamyE)-P glv .