响应面法优化产酰胺酶发酵培养基

Optimization of fermentation medium for amidase production by response surface methodology

采用响应面分析方法,对阿萨希丝孢酵母(Trichosporon asahii)ZZB-1产酰胺酶的发酵培养基进行了优化。运用单因子试验筛选出麦芽糖和酵母浸膏为最适碳源、氮源,金属离子Ca^(2+)、Mn^(2+)可提高发酵酰胺酶产量;通过最陡爬坡实验逼近以上4个因子的最大响应区域后,采用BoxBehnken响应面分析法,确定产酰胺酶最佳发酵培养基为麦芽糖18.84 g/L、酵母浸膏9.55 g/L、NaCl 5 g/L、KH_2PO_4 1 g/L、MgSO_4·7H_2O0.2 g/L、FeSO_40.001 g/L、CaCO_370.84μmol/L、MnSO_465.39μmol/L(1%丙烯酸诱导),NH_4·H_2O调节pH至7.0。培养基优化后酰胺酶产量由初始2554U/L提高到4156 U/L,为原始发酵培养基配方酶活产量的1.63倍。

Response surface methodology （RSM） was used to optimize fermentation medium for the total amidsae activity production of Trichosporon asahii ZZB-1 resting cells. Through the simple factor design of experiment , maltose and yeast extract were screened out as the most suitable carbon and nitrogen sources, meanwhile, the metal ions Ca2 ＋ , Mn2 ＋ had significant effects on amidase activity yield of resting cells. And the optimal concentrations of factors in medium were determined by response surface methodology after evaluating four factors related to the production of amidase activity. The results showed that the most suitable medium for amidase production was composed of maltose 18.84 g/L, yeast extract 9.55 g/L, NaCl 5g/L, KH2 PO4 1 g/L, MgSO4 7Ha O 0.2 g/L, FeSO4 0. 001 g/L, CaCO3 70.84μmol/L, MnSO4 65.39μmol/L, with 1% ammonium acrylate, which increased the yield of amidase activity from 2554 U/L to 4156 U/L.