一株高产蛋白酶菌株的筛选及其产酶条件-信息化建设中心-广东轻工.doc

一株高产蛋白酶菌株的筛选及其产酶条件( 林玩庄林淑娜Bacillus amyloliquefaciens)。通过摇瓶发酵试验，优选出可溶性淀粉和牛肉膏分别为最佳的碳源和氮源，并确定菌株PE11产蛋白酶的最佳条件为：温度30 °C、初始pH7.0、转速200 rpm和时间36 h。在最佳的产酶条件下，发酵液中的蛋白酶活力可达376 U/mL。 关键词：蛋白酶；高产；筛选；产酶条件 Study on screening and enzyme-producing conditions of a high protease producing strain LING Wan-zhuang, LING Shu-Na, CHEN Wen-cong, LIU Rong-lian, HUANG Ke-jia, HUANG Dan-min, XIE Gui-ren, CHEN Yu-bao, DENG Mao-cheng, WANG Yao, LI Jing (Guangdong Industry Technical College, Guangzhou 510300) Abstract：In order to improve the comprehensive utilization rate of protein resources from aquatic industry, strains having the ability to produce protease were isolated and screened from the gastrointestinal tract of large fish of South China Sea. Using flat transparent circle and shake flask fermentation test, a high producing protease strain PE11 was obtained. The strain PE11 was identified as Bacillus amyloliquefaciens through the systematic investigations of morphology, physiological and biochemical characteristics and 16S rDNA sequences analysis. By means of shake flask fermentation tests, the optimal carbon resource and nitrogen resource for strain PE11 were soluble starch and beef extract, respectively. In addition, the best conditions for protease-producing were determined as temperature of 30 °C, initial pH of 7.0 and rotation speed of 200 rpm. At the optimal condition, the highest protease activity of fermentation broth reached 376 U/mL. Key words：protease；high producing；screening；enzyme-producing condition 引言 近年来，我国的水产养殖量和加工量一直居世界首位[1-2]。但是，在捕捞和加工过程中，低值水产品往往占总产量的30%~50%[3-4]。如果这些低值水产品得不到高值化利用，将造成大量蛋白质资源的浪费。随着人们对低值水产品综合利用的重视，蛋白酶及其高产菌种的应用日益增加。为了直接利用高产蛋白酶菌种对低值水产品进行开发高值化产品，本实验主要筛选蛋白酶高产菌种，以及初步研究所得菌种的产酶条件。 1 材料与方法 1.1 筛选材料与试剂 筛选材料：南海海域大型鱼类的肠道组织；酪氨酸：分析纯，市售；Folin试剂及其它试剂：分析纯，市售。 1.2 培养基 富集培养基：葡萄糖 40 g/L，蛋白胨 10 g/L，酵母膏 5 g/L，Na2HPO4 2 g/L，KH2PO4 1 g/L，MgSO4·7H2O 0.5 g/L，pH7.0。分离平板培养基：葡萄糖 10 g/L，酵母膏 5 g/L，脱脂奶粉 10 g/L，NaCl 1 g/L，琼脂 10 g/L，pH7.0。斜面培养基：葡萄糖 10 g/L，蛋白胨 10 g/L，酵母膏 5 g/