摘要
为了获得高产中性蛋白酶芽孢杆菌菌株,试验从不同环境样本中分离细菌并进行16S rDNA序列鉴定,利用酪蛋白琼脂培养基对产蛋白酶菌株进行初筛、福林酚法检测中性蛋白酶活性,结合形态学和分子生物学鉴定对高产中性蛋白酶芽孢杆菌菌株进行分类鉴定,通过对培养时间、温度和pH值的调节,研究该菌株的酶学性质。结果表明:成功筛选出1株贝莱斯芽孢杆菌(Bacillus velezensis)A1,该菌株具有较强产中性蛋白酶能力,中性蛋白酶活性为31.40 U/mL,所产中性蛋白酶的最适反应温度为50℃、最适pH值为7,在pH值=6~8时稳定性较好,对碱性环境有一定的耐受性,5.0 mmol/L的Ca2+和Mn2+对中性蛋白酶活性有显著促进作用(P<0.05),Zn2+和Fe2+对中性蛋白酶活性有抑制作用(P<0.05),Mg2+无明显作用。说明贝莱斯芽孢杆菌A1可以作为生产中性蛋白酶的备选菌株。
In order to obtain high neutral protease-producing Bacillus strains, bacteria were isolated from different environmental samples and subjected to 16S rDNA sequence identification;high neutral protease-producing Bacillus strains were confirmed by primary screening using casein agar and re-screening for neutral protease activity by foline-phenol assay;the strains were classified and identified by combining morphological and molecular biological identification, and enzymatic properties were studied by adjusting incubation time, temperature and pH. The results showed that a strain of Bacillus velezensis A1 with strong neutral protease production ability was successfully screened, and the neutral protease activity was 31.40 U/mL;its optimum reaction temperature of neutral protease was 50℃;optimum pH value was 7, and the stability was good at pH=6-8, and it had a certain tolerance to alkaline environment. Ca2+and Mn2+at 5.0 mmol/L had a promotive effect on neutral protease activity;Zn2+and Fe2+had an inhibitory effect on neutral protease activity, and Mg2+had no significant effect.
作者
贾仲昕
赵佳男
季芳
王雪
李刚
王承民
秦建华
JIA Zhongxin;ZHAO Jianan;JI Fang;WANG Xue;LI Gang;WANG Chengmin;QIN Jianhua(College of Veterinary Medicine,Hebei Agricultural University,Baoding 071001,China;Guangdong Provincial Key Laboratory of Animal Conservation and Resource Utilization/Guangdong Provincial Public Laboratory of Wildlife Conservation and Utilization,Institute of Zoology,Guangdong Academy of Sciences,Guangzhou 510260,China)
出处
《黑龙江畜牧兽医》
CAS
北大核心
2023年第1期106-112,132,共8页
Heilongjiang Animal Science And veterinary Medicine
基金
河北省现代农业产业技术体系奶牛创新团队建设项目(HBCT2018120205)
河北省重点研发项目(20326603D)
广东省科学院人才专项(2016GDASRC-0205)
北京动物园圈养野生动物技术北京市重点实验室开放课题(ZDK202105)。
关键词
蛋白酶
芽孢杆菌
筛选
酶学性质
分离
鉴定
protease
Bacillus
screening
enzymatic properties
isolation
identification
