一株产碱性蛋白酶芽孢杆菌的筛选、鉴定及应用初探  

Screening,Identification and Application of a New Bacillus Producing Alkaline Protease

作　　者：徐雅芫[1] 李吕木[2,3] 许发芝[4,3] 丁小玲[4,3] 钱坤 

机构地区：[1]安徽农业大学生命科学学院,安徽合肥230036 [2]安徽农业大学生命科学学院动物科技学院,安徽合肥230036 [3]安徽省畜牧生物工程技术研究中心,安徽合肥230031 [4]安徽农业大学动物科技学院,安徽合肥230036

出　　处：《激光生物学报》2011年第6期830-837,823,共9页Acta Laser Biology Sinica

基　　金：国家科技人员服务企业项目(2009GJC30021);安徽省长三角联合科技攻关项目(10140702021);安徽省重大科技专项(07010301019)

摘　　要：为了筛选碱性蛋白酶产生菌并探讨其对蛋白质饲料的发酵效果,以肉粉厂表层土壤为菌株分离源,利用脱脂牛奶培养基分离和纯化蛋白酶产生菌,通过形态特征、生理生化和16S rRNA基因序列分析确定菌株的分类地位,并采用L_9（3~3）正交设计研究筛选出的优势菌种的接种量（3%、6%和12%）、种子液培养时间（12 h、24 h和48 h）和固态发酵时间（12 h、24 h和48 h）3因素对发酵豆粕和血粉的影响。此碱性蛋白酶产生菌与枯草芽孢杆菌亲缘关系最近,命名为枯草芽孢杆菌D-2。在最优条件下,该菌固态发酵豆粕及血粉可显著增加小肽含量至17.06%和12.73%;其最优条件为接种量6%、种子液培养时间24 h和发酵时间48 h。枯草芽孢杆菌D-2（Bacillus subtilis strain D-2）所产的碱性蛋白酶能高效降解豆粕和血粉中的蛋白,提高豆粕和血粉的饲用品质。To screen a bacterium producing alkaline protease and study its fermenting effect on protein feeds,a new strain was isolated from topsoil surface of meat power plant and purified using skim milk media;its taxonomy was established according to its morphological features,physiological and biochemical characteristics as well as phylogenetic analysis of 16S rRNA gene sequences;effects of inoculation,culture time of seed liquid and solid-state fermentation time on fermented performance of soybean meal and blood powder were investigated by orthogonal experiments.Results showed that this strain was the most closely related to Bacillus subtilis and named Bacillus subtilis strain D-2;under optional fermentation conditions,the contents of small peptides were significantly increased to 17.06%and 12.73%in fermented soybean meal and blood powder,respectively;these optional fermenting conditions as follows;the inculum of 6%,seed liquid culturing time of 24 h,and solid-state fermentation time of 48 h.These results indicated that alkaline protease produced by Bacillus subtilis strain D-2 effectively decomposed protein in both soybean meal and blood powder,which improved dietary performance of protein feeds.

关 键 词：碱性蛋白酶 枯草芽孢杆菌 分离鉴定 小肽 

分 类 号：Q939[生物学—微生物学]