高产酸性蛋白酶的菌株筛选及培养条件优化  

Screening of High-Yield Acid Protease Strains and Optimization of Culture Conditions

作　　者：何文彬 张捷文 徐晴元 侯莎 吴昌正 郭丽琼[1,2] 林俊芳 HE Wen-bin;ZHANG Jie-wen;XU Qing-yuan;HOU Sha;WU Chang-zheng;GUO Li-qiong;LIN Jun-fang(College of Food Science,South China Agricultural University,Guangzhou 510642,China;Guangdong Engineering Technology Research Center for Microecological Agents,Guangzhou 510642,China;Foshan Haitian(Gaoming)Flavouring&Food Co.,Ltd.,Foshan 528599,China)

机构地区：[1]华南农业大学食品学院,广州510642 [2]广东微生态制剂工程技术研究中心,广州510642 [3]佛山市海天(高明)调味食品有限公司,广东佛山528599

出　　处：《中国调味品》2025年第2期10-16,共7页China Condiment

基　　金：广东省调味食品生物发酵先进技术企业重点实验室开放基金项目(2017B030302002);广东省重点领域研究项目(2020B020226006,2022B0202050002)。

摘　　要：近年来,食品工业和饲料工业快速发展,企业对酸性蛋白酶的需求不断增长。发酵酱醪作为天然的微生物菌种库,含有各种特性优越的菌株。该研究旨在发酵酱醪中筛选出高产酸性蛋白酶的微生物菌株。共筛选出14株不同形态的霉菌,经过复筛,最后筛选出一株高产酸性蛋白酶的菌株黑曲霉(Aspergillus niger),命名为JL-08。随后,通过对黑曲霉JL-08产生的酸性蛋白酶进行性能评价,发现其具有在高盐发酵产品中的应用潜力。再采用单因素试验探索影响黑曲霉JL-08酸性蛋白酶酶活力的重要因素并利用Plackett-Burman试验筛选出碳氮比(C/N)、酪蛋白添加量、天冬氨酸添加量和苏氨酸添加量4个因素。最后进行四因素三水平的正交试验,确定了最优的因素组合为(C/N)16∶1、1%酪蛋白1.5 mL、30 mmol/L天冬氨酸1.5 mL。在此条件下,酸性蛋白酶酶活力提高至(5507±426)U/g。In recent years,the food and feed industries have developed rapidly,and the demand for acid protease from enterprises has been continuously increasing.Fermented sauce mash,as a natural microbial strain library,contains various strains with superior characteristics.The aim of this study is to screen high-yield acid protease microbial strains from fermented sauce mash.A total of 14 strains of molds with different morphologies are screened,and after re-screening,a high-yield acid protease strain Aspergillus niger is finally screened,which is named JL-08.Subsequently,the performance of acid protease produced by Aspergillus niger JL-08 is evaluated,and it is found that it has application potential in high-salt fermented products,and then through single factor test,the important factors that affect the activity of acid protease from Aspergillus niger JL-08 are explored,and Plackett-Burman test is used to screen four factors,including carbon-nitrogen ratio(C/N),casein addition amount,aspartic acid addition amount and threonine addition amount.Finally,four-factor and three-level orthogonal test is conducted to determine the optimal combination of factors:C/N of 16∶1,1%casein of 1.5 mL and 30 mmol/L aspartic acid of 1.5 mL.Under these conditions,the activity of acid protease increases to(5507±426)U/g.

关 键 词：酸性蛋白酶 筛选 黑曲霉 因素优化 

分 类 号：TS201.25[轻工技术与工程—食品科学]