为了提高重组大肠杆菌(ESCHErICHIA COlI)rOSETTA(dE3)-PET-22-bgl-(kIl-kM)的产酶能力,采用单因素试验研究了培养基碳源、氮源及碳氮比(摩尔比)对重组大肠杆菌rOSETTA(dE3)-PET-22-bgl-(kIl-kM)分泌表达重组β-1,3-1,4-葡聚糖酶H(A107-M)的影响.在此基础上,采用bOX-bEHnkEn设计法和响应面分析法对以上影响因素进行优化,得出最优培养基成分为(g/l):甘油10.01,酪蛋白胨12.03,酵母粉24,nACl 10,(nH4)2SO44.77,kH2PO42.31,k2HPO412.54.在In order to improve the enzyme activity of Escherichia coli Rosetta(DE3)-pET-22-bgl-(kil-Km),the effects of carbon sources,nitrogen sources and C/N ratio on the production of β-1,3-1,4-glucanase H(A107-M) by E.coli Rosetta(DE3)-pET-22-bgl-(kil-Km) were investigated by one-factor-a-time experiment.Box-Behnken design and a response surface methodology(RSM) were further used to optimize the above critical factors for enzyme production.The optimized medium contained(g/L):glycerol 10.01,casein peptone 12.03,yeast 24,NaCl 10,(NH4)2SO4 4.77,KH2PO4 2.31,and K2HPO4 12.54.When the recombinant E.coli was cultivated in the optimized medium for 27 h,the enzyme activity of β-1,3-1,4-glucanase H(A107-M) reached 78.32 U/mL(lichenan as the substrate),which was about 2.84 times of which obtained in the initial medium.科技部科研院所技术开发研究专项资金(NCSTE-2007-JKZX-023
