摘要
目的:通过石屏臭豆腐产蛋白酶优势菌株发酵获得高产蛋白酶菌种资源。方法:采用单因素和响应面试验对培养基及发酵条件进行优化;以酪蛋白为底物,利用酶学技术探究了温度、pH、金属离子、有机化学试剂对JX-11菌株蛋白酶活性和稳定性的影响。结果:从石屏臭豆腐中分离1株高产蛋白酶的优势菌JX-11,经鉴定为彭尼普尔金黄杆菌(Chryseobacterium pennipullorum)。JX-11菌株最优产酶条件为温度23.0℃、葡萄糖添加量6.7 g/L、蛋白胨添加量15.0 g/L、pH 6.4,该条件下所产蛋白酶酶活力为(39.16±3.24)U/mL。JX-11菌株产蛋白酶的最适温度为30℃,在10~40℃范围内具有良好的稳定性;最适pH为7.0,在pH 6.0~9.0范围内具有良好的稳定性;Mn 2^(+)可显著提高JX-11蛋白酶的活性,相对空白组酶活力提升了4.33倍;Zn 2^(+)、Cu 2^(+)、K^(+)均抑制该酶活性,Na^(+)、Mg 2^(+)、Ca 2^(+)无显著影响;甘油可以促进该酶的活性,乙醇、丙酮对酶活基本无影响,吐温80、乙酸、甲醇、EDTA抑制该酶活性,其中EDTA抑制作用最明显,进一步证实该酶为金属蛋白酶类。结论:从石屏臭豆腐中获得一株高产蛋白酶菌株,具有较好的应用前景。
Objective:Obtaining hhigh-yield pprotease-pproducing bacterial resources through fermentation of advantageous protease-producing strains derived from Shiping stinky tofu.Methods:Single-factor and response surface experiments were used to optimize the culture medium and fermentation conditions.Using casein as the substrate,the effects of temperature,pH,metal ions and organic reagent on the protease activity and stability of strain JX-11 were investigated using enzymology techniques.Results:One strain of high-producing proteinase-secreting bacteria,JX-11,was isolated from Shiping stinky tofu,identified as Chryseobacterium pennipullorum.The optimal conditions for protease production by JX-11 strain were found to be temperature at 23.0℃,glucose content at 6.7 g/L,peptone content at 15.0 g/L,pH 6.4.Under these conditions,the protease activity was(39.16±3.24)U/mL.The optimal temperature for extracellular protease of JX-11 strain was 30℃,and it had good stability in the range of 10~40℃.The optimal pH was 7.0,and it had good stability in the range of pH 6.0~9.0.Mn 2^(+)significantly increased the activity of JX-11 protease.The relative activity was increased by 4.33 times compared with the blank group.Zn 2^(+),Cu 2^(+),and K^(+)all inhibited the enzyme activity,Na^(+),Mg 2^(+),and Ca 2^(+)had no significant effect.Glycerol can promote the activity of the enzyme,but ethanol and acetone have little effect on it.Tween 80,acetic acid,methanol and EDTA inhibited the activity of the enzyme,among which EDTA hadd the most obvious inhibition,which further proved that the enzyme was a metalloprotease.Conclusion:A high-producing protease-secreting bacterial strain is obtained from Shiping stinky tofu,which has good application prospects.
作者
刘超
赵良忠
王瑶琼
李明
冯绪忠
LIU Chao;ZHAO Liangzhong;WANG Yaoqiong;LI Ming;FENG Xuzhong(College of Food Science and Chemical,Shaoyang University,Shaoyang,Hunan 422000,China;Key Laboratory of Bean Products Processing and Safety Control,Shaoyang,Hunan 422000,China;Guangzhou Jiaming Food Technology Co.,Ltd.,Guangzhou,Guangdong 511458,China;Shenzhen Shanggutang Food Development Co.,Ltd.,Shenzhen,Guangdong 518000,China)
出处
《食品与机械》
CSCD
北大核心
2024年第3期33-43,共11页
Food and Machinery
基金
邵阳学院研究生科研创新项目(编号:CX2022SY014)
湖南省研究生科研创新项目(编号:QL20220251)
湖南省大学生创新创业训练计划项目(编号:S202210547066)
湖南省科技创新计划资助项目(编号:2022NK2039)。
关键词
彭尼普尔金黄杆菌
蛋白酶
筛选鉴定
酶学性质
Chryseobacterium pennipullorum
protease
screening and identification
enzymatic properties
作者简介
刘超,女,邵阳学院在读硕士研究生;通信作者:赵良忠(1963—),男,邵阳学院教授,硕士。E-mail:sys169@163.com。
