| 溶解性多糖单加氧酶CBP21的高效分泌表达及与几丁质酶的协同作用研究 |
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| 引用本文： | 李志敏§,潘兴亮§,杨雅麟,刘智,徐俐,何夙旭,周志刚.溶解性多糖单加氧酶CBP21的高效分泌表达及与几丁质酶的协同作用研究[J].中国农业科技导报,2017,19(1):58-65. |
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| 作者姓名： | 李志敏§ 潘兴亮§ 杨雅麟 刘智 徐俐 何夙旭 周志刚 |
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| 作者单位： | 中国农业科学院饲料研究所, 农业部饲料生物技术重点实验室, 北京 100081 |
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| 基金项目： | 海洋公益性行业科研专项经费项目(201305022-6)； 国家973计划项目（2015CB150600）；北京市鲟鱼、鲑鳟鱼产业技术体系项目(SCGWZJ20141104-4)资助。 |
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| 摘 要： | 为实现溶解性多糖单加氧酶CBP21在枯草芽孢杆菌中的高效分泌表达,并对其与几丁质酶的协同作用进行研究,以大肠-枯草芽孢杆菌穿梭质粒p WB980为基本骨架,溶解性多糖单加氧酶为目的蛋白,构建了信号肽筛选载体,依据信号肽结构特征(正电荷与疏水性的差异)对Sec途径的13个信号肽进行了筛选,结果显示信号肽Ylq B引导分泌溶解性多糖单加氧酶的效率最高,其发酵液上清对胶体几丁质结合活力最高,上清中结合蛋白占总蛋白浓度为32.39%。进一步分别对CBP21和来自糖苷水解酶18家族粘质沙雷菌几丁质酶及来自糖苷水解酶19家族的嗜水气单胞菌几丁质酶Chi92的协同作用进行分析,表明CBP21对几丁质酶和Chi92活力均有促进作用,其中使粘质沙雷菌几丁质酶活力提高1.85倍,使Chi92(嗜水气单胞菌)活力提高2.35倍。上述研究为该溶解性多糖单加氧酶的工业应用奠定了基础。
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| 关 键 词： | /> 溶解性多糖单加氧酶 枯草芽孢杆菌 信号肽 协同效应 |
Studies on the Efficient Secretion of Lytic Polysaccharide Monooxygenases CBP21 and its Synergism with Chitinase |
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| LI Zhi-min§,PAN Xinliang§,YANG Yalin,LIU Zhi,XU Li,HE Suxu,ZHOU Zhigang.Studies on the Efficient Secretion of Lytic Polysaccharide Monooxygenases CBP21 and its Synergism with Chitinase[J].Journal of Agricultural Science and Technology,2017,19(1):58-65. |
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| Authors: | LI Zhi-min§ PAN Xinliang§ YANG Yalin LIU Zhi XU Li HE Suxu ZHOU Zhigang |
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| Institution: | Key Laboratory for Feed Biotechnology, Ministry of Agriculture; Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China |
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| Abstract: | In order to achieve the efficient secretion expression of lytic polysaccharide monooxygenases CBP21 in B. Subtilis, and investigate the synergistic effect of CBP21 with chitinase, this experiment constructed a series of screening vectors for signal peptides from B.subtilis. The CBP21 was cloned into Escherichia coli-Bacillus Subtilis shuttle vector pWB980, and 13 Sec-pathway signal peptides with different structure were chosen. The results showed that the signal peptide YlqB could secret the CBP21 better than the other signal peptides. The bound protein accounted for 32.39% of the total protein in supernatant, showing the highest binding activity for colloidal chitin. Furthermore, the degradation of colloidal chitin with Serratia marcescens chitinase and Aeromonas hydrophila Chi92 in the presence of CBP21 were determined. The result indicated that the addition of CBP21 could increase both the activities of S. marcescens chitinase and A. hydrophila Chi92. The activity of Chi92 was improved by 2.35 fold. Whereas for S. marcescens chitinase, CBP21 increased its activity by 1.85 fold. This study laid solid foundation for its industrial application. |
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| Keywords: | lytic polysaccharide monooxygenases Bacillus Subtilis signal peptide synergistic effect |
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