土壤中产纤维素酶菌株的发酵条件及酶学性质的研究  

Study on the fermentation conditions and enzymatic properties by a high-yield cellulase strain from soil

作　　者：曹慧[1] 杨佳萌 张腾月 张会会 王梦瑶 任世威 张朋振 叶京飞 CAO Hui;YANG Jia-meng;ZHANG Teng-yue;ZHANG Hui-hui;WANG Meng-yao;REN Shi-wei;ZHANG Peng-zhen;YE Jing-fei

机构地区：[1]河南科技大学动物科技学院,河南洛阳471000 [2]广东海洋大学农学院,广东湛江524088

出　　处：《饲料研究》2021年第5期63-69,共7页Feed Research

基　　金：河南科技大学博士科研启动资金(项目编号:13480077);青海省重点研发计划(项目编号:2017-NK-153、2019-NK-173、2019-SF-149)。

摘　　要：试验探究从青藏高原土壤中筛选出的高产纤维素酶菌株产酶能力,并对高产纤维素酶菌株进行发酵条件优化以及酶学特性研究。采用刚果红染色法(水解圈法)和3,5-二硝基水杨酸(DNS)法考察菌株产纤维素酶能力,采用形态学和分子生物学进行菌种鉴定,采用单因素试验和正交试验探究发酵条件优化及酶学性质。结果显示,筛选分离出一株高产纤维素酶菌株SD10,经鉴定为粗糙脉孢菌(Neurospora crassa)。菌株SD10最佳发酵产酶条件为:玉米秸秆5%、豆粉1%、Mn2+0.2%、培养温度23℃、初始pH值8.0、接种量2%的条件下200 r/min振荡培养84 h,产纤维素酶活力最高为9.18 U/mL。菌株SD10纤维素酶最适反应温度50℃,最适pH值4.0。在40~50℃、pH值5.0~6.0条件下,酶活稳定性较好。金属离子中对纤维素酶活力有明显激活作用的是Mn2+,有明显抑制作用的是Fe3+、Zn2+、Cu2+。试验表明,粗糙脉孢菌SD10以玉米秸秆为碳源产生的酸性纤维素酶具有良好的温度稳定性和pH值稳定性,具有潜在的开发利用价值。To screen the strain with high cellulase activity from the alpine meadow soil on the Tibetan plateau, optimize fermentation conditions and characterize the enzyme, which lay a theoretical foundation for further development and utilization. Hydrolyzing ring method and 3,5-Dinitrosalicylic acid(DNS) method were used to screen strains. Strains were identified by morphology and molecular biological identification. The fermentation condition and enzymatic properties were conducted by single factor and/or orthogonal test. A strain named SD10 with high cellulase activity was identified as Neurospora crassa. The optimum fermentation conditions were corn straw 5%, soybean flour 1%, Mn2+0.2%, 23 ℃, pH value 8.0, inoculum amount 2%, rotation speed 200 r/min after 84 h. The highest cellulase activity of SD10 reached9.18 U/mL by optimization. The optimum reaction temperature and pH value of the cellulase was 50 ℃ and 4.0 respectively. The enzyme activity is relative stable under 40~50 ℃ or pH value 5.0~6.0. The cellulase activity of SD10 was significantly enhanced by Mn2+, but significantly reduced by Zn2+, Cu2+and Fe3+. Neurospora crassa SD10 can produce acid cellulase with good temperature stability and pH value stability by using agricultural by-product(corn straw), which has potential development value.

关 键 词：纤维素酶 筛选鉴定 发酵条件优化 酶学特性 青藏高原 

分 类 号：S816.3[农业科学—饲料科学]