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2010 No.5 Serial No.218
China Brewing
Research Report
郸县豆瓣曲曲霉的筛选、分子鉴定及其酶学性质的研究
李从虎”，郑佳，谢菲"，陈欣”，周荣清12*
(1.四川大学轻纺与食品学院，四川成都610065;2.四川大学制革清洁技术国家工程实验室，四川成都610065)
摘要：以自然接种的期曲为研究对象，应用示踪和基于基质特性组合的菌种筛选技术，筛选出适合其酿造特性的分离株No17。通过形态学观察和26SrDNA、ITS序列的分子生物学鉴定，该分离株为Aspergillusoryzae，命名为A.oryzaeSZP17。该菌株的中性蛋白酶和α-淀粉酶的酶学性质研究表明，中性蛋白酶的最适反应温度为40℃，20℃~40℃时比较稳定，最适反应pH值为7.5，耐酸性较强，Cu* 和Mn*对其具有激活作用，Fe*和Fe*对其具有抑制作用。α-淀粉酶的最适反应温度为60℃，低于50C比较稳定，最适反应pH值为6.0，
pH值>7.5时易失活，各种金属离子对其酶活影响较小。关键词：筛选：鉴定：郭县豆摄婚曲；A.oryzae:酶学性质
中图分类号：Q93-331
文献标识码：A
文章编号：02545071(2010)05003005
Screening,molecular classification of Aspergillus in Pixian'horse-bean and its enzymatic characteristics
LI Conghu', ZHENG Jia', XIE Fei', CHEN Xin', ZHOU Rongqingl-2*
(1.National Engineering Laboratory for Clean Technology of Leather Manufacture, Sichuan University, Chengdu 610065, China;
2. Textile Industry & Food Science College of Sichuan University, Chengdu 610065, China)
Abstract: The flora of naturally cultured Pixian'horse-bean koji was anatomized. By screening technique of integrating the tracer with the culture substrates characteristics, strain No17 was obtained. The isolate was identified belonging to Aspergillus oryzac according to the results of morpholog) and molecular classification of 26S rDNA and ITS, and named as ALoryzae SZP17. The enzymatic characteristics related to brewing properties such as neutral protease and a-amylase were measured. The optimal temperature and pH for neutral protease were 40°C and 7.5, respectively. The neutral protease remained better stability between 20°C40°C, and had strong acid-resistance, stimulated by Cu*" and Zn*, whereas inhibited by Fe* and Fe*. The optimal temperature and pH for a-amylase were 60°C and 6.0, respectively. The a-amylase remained better stability under 50°C, and metal ions used in present study had less effects on the activities.
Key words: screening; identification; Pixian'horse-bean koji; A.oryzac, enzymatic characteristics
具有300多年酿造历史的郏县豆以其味辣香醇、红棕油亮、粘稠绒实、酱香浓郁的风格在我国的传统酿造酱类中独树一帜，塔称川菜之魂。产品的特色源于自然接种、生料制曲的独特酿造工艺和自然环境。在长期使用的生产环境中，蕴藏着颇具特色的微生物资源，是认识和掌握
收稿日期：2009-11-18
基金项目：四川省重点科技项目(07SG111-009)
微生物作用规律的重要研究客体。部析、认识郸县豆需需曲的微生物菌群特征，是弘扬传统酿造工艺特点，实现集约化、规模化生产的重要基础。
中性蛋白酶和α-淀粉酶在工业生产中具有极其重要的地位，是工业生产中用途最广的酶制剂之一。其作用条件
作者篇介：李从虎(1987-)，男，安徽东至人，在读硕士研究生，研究方向为现代发酵技术；周荣清*，教授，通讯作者。
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