| Trichoderma lentiforme ACCC30425脂肪酶基因的原核表达及其酶学性质的初步研究 |
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| 作者姓名： | 王宇洲 马锐 向杰 陈敬师 李世贵 龚明波 顾金刚 |
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| 作者单位： | 1.中国农业科学院农业资源与农业区划研究所, 农业农村部农业微生物资源收集保藏重点实验室, 北京 100081;
2.中国农业科学院饲料研究所, 农业农村部饲料生物技术重点实验室, 北京 100081 |
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| 基金项目： | 国家科技基础性工作专项资金项目（2014FY120900）；中央级公益性科研院所基本科研业务费专项资金（IARRP-2015-24；1610132016050）资助。 |
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| 摘 要： | 基于在Trichoderma lentiforme ACCC30425全基因组序列注释的15个脂肪酶基因，进行脂肪酶基因合成与Escherichia coli BL21异源表达，获得11个产脂肪酶阳性克隆，其中酶活力最高为1.5 U/mL。经初步酶学分析，结果表明重组表达脂肪酶普遍具有耐热性、耐碱性的特征，最适温度范围为50~90℃，最适pH范围为8.5~9.5。其中，LipP最适温度为90℃，远高于目前已知脂肪酶的最适温度；LipQ最适温度为50℃，最接近洗涤工业的中温温度；其他脂肪酶最适温度为70℃或80℃，均属耐热脂肪酶。
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| 关 键 词： | 木霉 脂肪酶 原核表达 耐热性 |
| 收稿时间： | 2018-03-22 |
Prokaryotic Expression of Recombinant Lipases Gene from Trichoderma lentiforme ACCC30425 and Preliminary Study of its Enzymatic Properties |
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| Authors: | WANG Yuzhou MA Rui XIANG Jie CHEN Jingshi LI Shigui GONG Mingbo GU Jingang |
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| Institution: | 1.Key Laboratory of Microbial Resources Collection and Preservation, Ministry of Agriculture and Rural Affairs, Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, Beijing 100081, China;
2.Key Laboratory of Feed Biotechnology, Ministry of Agriculture and Rural Affairs, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China |
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| Abstract: | According to the genome analysis of Trichoderma lentiforme ACCC30425, the coding sequences of 15 lipase genes were annotated, and then used for gene synthesis and heterologous expression in Escherichia coli BL21. 11 lipase-producing clones were obtained, and the highest lipase activity was up to 1.5 U/mL. Results of preliminary enzymatic analysis indicated that the recombinant lipases had the characteristics of heat resistance and alkali resistance universally, showing optimal activities at 50~90℃ and pH 8.5~9.5. Among them, the optimum temperature of LipP was 90℃, which was much higher than all known counterparts, and LipQ showed optimal activity at 50℃, which was close to the intermediate temperature of detergent industry. And other lipases were also thermostable lipase, showing optimal activities at 70℃ or 80℃. |
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| Keywords: | Trichoderma lipase prokaryotic expression heat resistance |
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