天祝放牧牦牛生活环境土壤中一株产凝乳酶细菌的分离与鉴定 - 食品科学.pdfVIP

158 2009, Vol. 30, No. 11 食品科学 ※生物工程 天祝放牧牦牛生活环境土壤中一株产凝乳酶 细菌的分离与鉴定 宋 曦，甘伯中 * ，贺晓玲，张卫兵，李 帆，刘兴龙 (甘肃农业大学食品科学与工程学院，甘肃农业大学研究测试中心，甘肃 兰州 730070) 摘 要：筛选高产凝乳酶细菌，并对其进行鉴定和产凝乳酶活力测定。利用酪蛋白培养基从天祝放牧牦牛生活环 境土壤中筛选高产凝乳酶细菌，利用形态观察、生理生化检测、16S rDNA 序列分析对其进行鉴定，并采用 Arima 法和福林法分别测定凝乳酶活力和蛋白水解活力。获得 18 株产凝乳酶细菌，其中菌株 D3.11 产凝乳酶活力相对最 高，鉴定为地衣芽孢杆菌(Bacillus licheniformis) ，其在麸皮培养基中发酵72 h 产凝乳酶活力高达 89.56 SU/ml，蛋 白水解力为21.27U/ml 。菌株D3.11 是一株高产凝乳酶细菌，可作为候选菌株进行进一步研究。 关键词：凝乳酶；细菌；分离鉴定； 16S rDNA ； 地衣芽孢杆菌 Separation and Identification of A Chymosin Producing Bacterium from Soil of Yak Grazing District in Tianzhu Country of Gansu Province SONG Xi，GAN Bo-zhong* ，HE Xiao-ling ，ZHANG Wei-bing ，LI Fan ，LIU Xing-long (College of Food Science and Engineering, Gansu Agricultural University, Analysis and Research Center of Gansu Agricultural University, Lanzhou 730070, China) Abstract ：An efficient chymosin producing strain, D3.11, was screened out from 18 bacterial strains isolated from the soil of yak grazing district in Tianzhu County of Gansu Province, China, using casein medium, whose chymosin and proteolysis activities were 89.56 SU/ml and 21.27 U/ml after 72 h shaking fermentation (140 r/min) at 37 ℃ in bran medium, respectively. This strain was identified as Bacillus licheniformis by the means of bacterial morphological feature observation, physiological and biochemical characteristics measurement and 16S rDNA sequence analysis. Furthermore, the growth and enzyme production curves of this strain were plotted. Key words ：chymosin ；bacterium ；separation and identification ；16S rDNA；Bacillus licheniformis 中图分类号：Q93-331 文献标识码：A 文章编号：1002-6630(2009)11-0158-