Optimization of cultural conditions for thermostable β-1, 3-1, 4-glucanase production byBacillus subtilis ZJF-1A5

Abstract

The optimization of cultural conditions for β-glucanase production byBacillus subtilis ZJF-1A5 was investigated in flask trials. Temperature had great effect on β-glucanase production which maximized at optimal temperature of 37°C and decreased significantly when temperature was over 37°C. Charge quantity affected β-glucanase production significantly. Adding oxygen vector N-dodecane or acetic ether benefited β-glucanase production, but it depended on the concentration and charge quantity. The results of fractional factorial design showed that age and size of inoculum and shaking speed were the key factors affecting β-glucanase production and the cultivation time span to reach the highest β-glucanase activity. The optimal cultural conditions for β-glucanase production obtained with CCD were as follows: inoculum age and size (16 h, 3.82% (v/v)), shaking speed 210 r/min, charge quantity of 30 mL in 250 mL flask and initial pH 7.0, cultured at 37°C for 50 h. Repeated experimental results accorded with those predicted by a second-order polynomial model. The amount of β-glucanase, α-amylase and neutral protease produced byB subtilis ZJF-1A5 was associated partially with cell growth. Those three enzymes' activities increased following the cell growth and increased significantly when cells entered the stationary phase.