高活力碱性蛋白酶产生菌的选育与发酵放大  

Selection and fermentation scale-up of high-activity alkaline protease

作　　者：孙同毅[1] 殷向斌[1] 郝继兴[1] 陈坤[1] 路福平[1] 杜连祥[1] 

机构地区：[1]天津市工业微生物重点实验室 天津科技大学生物工程学院,天津300457

出　　处：《辽宁工程技术大学学报（自然科学版）》2009年第4期679-682,共4页Journal of Liaoning Technical University (Natural Science)

基　　金：国家高技术研究发展计划(863计划)基金资助项目(2007AA02Z212);国家科技资源平台基金项目资助(2005DKA21204-10)

摘　　要：为了选育高活力碱性蛋白酶产生菌株。采用复合诱变(紫外照射、NTG、离子注入)方法,结合平板初筛与摇瓶复筛。获得一株高产突变株HAPN-169,其酶活力为3.5×10u/mL。高产菌株HAPN-169根据气液接触中体积传氧系数kLa相同的原则发酵放大(7L,30L,200L),其发酵参数为:温度34℃,装液系数0.7,接种量5%,使溶氧维持在20～30%,控制发酵过程pH不低于7.0。在7L发酵罐,发酵时间为54小时,酶活力为3.6×104u/mL;在30L发酵罐,发酵时间为50小时,酶活力为3.86×104u/mL;在200L发酵罐,发酵时间为36小时,酶活力为4.2×104u/mL。发酵液pH在7.5左右时,酶活力在峰值,发酵终点一到,必须马上下罐。该结果对为我国碱性蛋白酶工业发展提供一定的基础。In order to screen high-yield alkaline protease producing strain, the complex mutation （UV radiation, NTG, and N^＋ ion implantation） was applied and the plate screen and shake flask screen were carried out in the study. The high-yield protease strain HAPN-169 was obtained with its protease activity as 3.5×10^4u/mL. The experiment was then scaled up into 7L, 30L and 200L fermentors on the basis of same kLa. The optimum fermentation conditions were obtained as： 1） the optimal culture condition is 34℃; 2） the fermentation medium volume is 0.7; 3） inoculation is 5%; 4） dissolved oxygen is approximately 20-30%; and 5） the pH value is over 7.0 during the controlled fermentation process. In a 7L fermentor, the alkaline protease activity was about 3.6× 10^4 u/mL after cultivating for 54 hours. In a 30L fermentor, the protease activity was about 3.86×10^4 u/mL after cultivating 50 hours. In a 200L fermentor, the protease activity was about 4.2× 10^4u/mL after cultivating 36 hours. When the pH-value of the culture was around 7.5, the activity of alkaline protease was close to tip, and the fermentation must be stopped. The study laid a solid basis for the development of alkaline protease industry.

关 键 词：碱性蛋白酶 离子注入 发酵放大 发酵终点 

分 类 号：O629[理学—有机化学]