高产纤溶酶菌株筛选及发酵鹰嘴豆培养基优化  

Screening of strain with high production of fibrinolytic enzyme and optimization of medium composition by using chickpea as substate

作　　者：肖萍[1] 姚四平[1] 康然[1] 刘艳琦[1] 王艳萍[1] 

机构地区：[1]食品营养与安全教育部重点实验室,天津科技大学食品工程与生物技术学院,天津300457

出　　处：《食品科技》2014年第10期32-38,共7页Food Science and Technology

基　　金：国家科技部863课题(2013AA102204)

摘　　要：从大豆发酵物中分离出一株高产纤溶酶的菌株,经API实验及16S rDNA序列分析鉴定该菌株为枯草芽孢杆菌(Bacillus subtilis)。并对该菌株发酵产生的纤溶酶性质进行了初步研究,表明该菌株产生的纤溶酶为碱性丝氨酸金属蛋白酶,并且具有直接水解纤维蛋白和激活纤溶酶原的作用,为一种新型的纤溶酶。同时,为了提高鹰嘴豆的经济价值,首次采用液态发酵方法利用筛选得到的菌株发酵鹰嘴豆产纤溶酶,通过Plackett-Burman设计和响应面结合的方法,确定其最优培养基组分(质量浓度)为:蔗糖50.0 g/L,鹰嘴豆34.85 g/L,氯化钙0.42 g/L,硫酸镁0.35 g/L,磷酸氢二钾5.00 g/L,磷酸二氢钾6.00 g/L。优化后枯草芽孢杆菌发酵鹰嘴豆产纤溶酶活力为3210U/mL,比基础培养基提高了2.01倍。In this experiment one high producing fibrinolytic enzyme strain was screened from the fermented soybean and identified. It was identified as Bacillus subtilis according with the API experiment and 16S rDNA sequences. Furthermore, the fibrinloytic enzyme biochemical characterization was also evaluated. The results showed that the fibrinloytic enzyme was an alkaline serine metalloprotease while having both plasmin and plasminogen activator-like activities. In order to promote the economic value of the chickpea, liquid fermented of chickpeas with isolated Bacillus subtilis was first investigated. The culture medium for enzyme producing was optimized by Plackett-Burman and Box-Behnken design and the optimal fermentation medium was composed of （in g/L）： sucrose 50.0, chickpea flour 34.85, calcium chloride 0.42, magnesium sulfate 0.35, dipotassium hydrogen phosphate 5.00, potassium dihydrogen phosphate 6.00. The activity of the Bacillus subtilis to produce fibrinolytic enzyme was 3210 U/mL after optimization, which was 2.01 folds higher than that of basal medium.

关 键 词：鹰嘴豆 液态发酵 纤溶酶 培养基优化 

分 类 号：TS201.3[轻工技术与工程—食品科学]