响应面-满意度函数优化重组大肠杆菌产NMN转移酶发酵条件  

Optimization of NMN adenylyltransferase production by recombinant Escherichia coli using response surface methodology coupled with desirability function

作　　者：段琳琳[1] 李红梅[1] 何海[1] 

机构地区：[1]上海理工大学医疗器械与食品学院,上海杨浦200093

出　　处：《工业微生物》2016年第6期47-53,共7页Industrial Microbiology

摘　　要：重组大肠杆菌Escherichaia coli能高效表达NMN转移酶,以此为出发菌株,以菌体生长量OD600和NMN转移酶的活力为响应值,对重组大肠杆菌产NMN转移酶的发酵条件进行优化。首先以Plackett-Burman实验设计优化筛选出3个主要影响因子:胰蛋白胨、甘油、Mg SO4;随后以BoxBehnken中心组合设计建立上述3个因子对OD600和NMN转移酶活力水平的数学模型;最后通过满意度函数获得最佳发酵条件为:酵母粉30 g/L,胰蛋白胨10.5 g/L,甘油3.49 m L/L,Mg SO40.45g/L,K2HPO440.5 g/L,KH2PO46.0 g/L,NH4Cl 1.5 g/L,Na Cl 0.6 g/L,接种量1.5%,诱导时间12h。在该优化条件下,菌体生长和产酶水平均获得了显著的提升。重组NMN转移酶的活力水平从8.85 U/mg提高到15.48 U/mg,菌体生长量OD600从4.85提高到6.01,提高幅度分别为74.92%和23.92%。The fermentation conditions of NMN adenylyltransferase production by recombinant Escherichia coli were optimized. Biomass yield （OD600 ） and NMN adenylyhransferase activity were selected as response value during fermentation condition optimization. Peptone, glycerin and MgSO4 were screened to be the three key components using Plackett-Burman design method. Then, the mathematical model reflecting the three main factors and the production level of enzyme was established by using the Box-Behnken design method. Finally, a satisfactory degree function was defined and the satisfactory solution was obtained. The final optimal fermentation conditions were： yeast extract 30 g/L, peptone 10.5 g/L, glycerol 3.49 mL/L, MgSO4 0.45 g/L, K2HPO4 40.5 g/L, KH2PO4 6.0 g/L, NH4Cl1.5 g/L, NaCl 0.6 g/L, inoculum size of 1.5% and induction time of 12 h. After the validated experiment under above conditions, enzyme production and biomass of recombinant strain were increased by 74.92% and 23.92%, respectively.

关 键 词：NMN转移酶 发酵优化 满意度函数 响应面法 

分 类 号：TQ920.6[轻工技术与工程—发酵工程] Q93[生物学—微生物学]