响应面分析优化产木聚糖酶毕赤酵母基因工程菌培养条件  

Response surface optimization of fermentation conditions for xylanase production by recombinant Pichia pastoris

作　　者：郭宁[1] 赵伟[1,2] 郑甲[1] 田健[1] 吴丽双[1] 周洪波[1,2] 

机构地区：[1]中南大学资源加工与生物工程学院,长沙410083 [2]生物冶金教育部重点实验室,长沙410083

出　　处：《工业微生物》2012年第3期55-60,共6页Industrial Microbiology

基　　金：教育部新世纪优秀人才支持计划(No.NECT-06-0691)

摘　　要：研究不同碳源、氮源和无机盐对毕赤酵母AX181菌株产木聚糖酶的影响。实验表明,分别采用葡萄糖和玉米浆干粉为碳源和氮源可以明显提高木聚糖酶的产量。无机盐单因子优化实验显示添加适量的(NH_4)_2SO_4、KH_2PO_4、MnSO_4·H_2O、FeSO_4·7H_2O也可以部分提高木聚糖酶产量。在此基础上利用响应面法优化毕赤酵母产木聚糖酶培养基,利用12次实验的Plackett-Burman设计实验筛选出影响产木聚糖酶的3个主要因素,即玉米浆干粉、MnSO_4·H_2O和FeSO_4·7H_2O。并进一步通过最陡爬坡路径逼近最大响应区域,采用中心组合实验设计确定最佳条件。优化后的产木聚糖酶培养基组分为(g/L):葡萄糖40.00,玉米浆干粉80.84,(NH_4)_2SO_4 6.25,KH_2PO_4 1.25、MnSO_4·H_2O 0.35,FeSO_4·7H_2O 1.31。培养基优化后,实际产酶2 883.86 U/mL,是优化前YPD培养基产酶的2.51倍。The effects of carbon sources, nitrogen sources and inorganic salts on improving quantity of xylanase produced by recombinant Pichia pastoris strain Ax181 were studied. The results showed that the cheap and facile glucose and com steep liquor powder obviously enhanced the production of xylanase. Single factor experiment of inorganic salts indicated that proper concentration of （NH4）2 SO4, KH2 PO4, MnSO4 ·H2O and FeSO4· 7H2O also improved xylanase production. Then, an optimization of culture conditions was carried out based on response surface methodology （RSM）. Three variables i.e. corn steep liquor powder, MnSO4· H2O and FeSO4 ·7H2O were identified to significantly affect xylanase production by Plackett-Burman design. The path of steepest ascent was applied to approach the optimal region. Central composite design （CCD） and response surface analysis were used to further investigate the optimal levels and relationships of these three factors. The optimal medium composition （g/L） estimated for maximum of xylanase production were： glucose 40.00, corn steep liquor powder 80.84, （NH4）2SO4 6.25, KH2PO4 1.25, MnSO4 · H2O 0.35, and FeSO4 · 7H2O 1.31. After medium optimization, the xylanase production reached 2 883.86 U/mL, which was 2.51-fold of that using YPD medium.

关 键 词：响应面分析 木聚糖酶 培养基优化 毕赤酵母 

分 类 号：TQ925[轻工技术与工程—发酵工程]