漆酶N端融合His-tag或S-tag标签促进其在大肠杆菌中可溶性表达

doi:10.13560/j.cnki.biotech.bull.1985.2016.03.028

摘要/Abstract

摘要： 来源于地衣芽孢杆菌的漆酶具有催效率高, 底物范围广等优点在工农业等领域具有重要的应用前景, 但该酶在外源基因表达系统中的表达量低, 影响了其在工农业等领域的应用。His-tag和S-tag两种标签由于具有分子量小, 且一般不影响外源蛋白性质等特点, 在外源蛋白的纯化和检测中具有重要的应用。本研究发现将来源于地衣芽孢杆菌的漆酶CotA的N端融合His-tag或S-tag构建于pET-22b载体, 并转化大肠杆菌BL21(DE3)后可以显著提高其在大肠杆菌中的可溶性表达量, 其表达量较N端无融合标签的构建, His-tag可提高漆酶表达量约为37倍, S-tag约可提高20倍, His-tag与S-tag共作用约可提高28倍。

关键词: 漆酶, His-tag, S-tag, 高效表达

Abstract: Laccase from Bacillus licheniformis has the advantages of high catalytic efficiency, wide range of substrates, etc., thus it owns the broad application prospect in industrial and agricultural fields. However, the expression level of the enzyme in the foreign gene expression system is low, which greatly limits its application in the agricultural and industrial fields. His-tag or S-tag is the small peptide with low molecular weight and usually can not affect the characteristics of heterologous fusion protein, and they are beneficial to be applied in the purification and detection of heterologous proteins. In this study, fusion of His-tag and/or S-tag on the N-terminal of the laccase CotA from B. licheniformis was constructed into pET-22b vector, and the vector was transferred into Escherichia coli BL(DE3), then we found that the soluble expression level of laccase significantly increased in E. coli BL(DE3). Compared with the construction without no fusion of any tags on N-terminal, the expression level with His-tag was about 37 folds, 20 folds with S-tag, and 28 folds with both His-tag and S-tag, respectively.

Key words: laccase, His-tag, S-tag, high expression

岳青霞,张丽洁,田健,伍宁丰,姚冬生. 漆酶N端融合His-tag或S-tag标签促进其在大肠杆菌中可溶性表达[J]. 生物技术通报, 2016, 32(3): 178-183.

YUE Qing-xia, ZHANG Li-jie, TIAN Jian, WU Ning-feng, YAO Dong-sheng. Promotion of Soluble Expression of Laccase in Escherichia coli by Fusion of His-tag or S-tag on the N-terminal of It[J]. Biotechnology Bulletin, 2016, 32(3): 178-183.

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https://biotech.aiijournal.com/CN/Y2016/V32/I3/178

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漆酶N端融合His-tag或S-tag标签促进其在大肠杆菌中可溶性表达

Promotion of Soluble Expression of Laccase in Escherichia coli by Fusion of His-tag or S-tag on the N-terminal of It

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[1]	王雨辰, 丁尊丹, 关菲菲, 田健, 刘国安, 伍宁丰. 耐热漆酶ba4基因鉴定与酶学性质分析[J]. 生物技术通报, 2022, 38(8): 252-260.
[2]	贾晨波, 苏一黄, 马秀梅, 王春利, 徐春燕. 端梗霉Z45产漆酶培养基的优化及其对染料的脱色[J]. 生物技术通报, 2022, 38(6): 252-260.
[3]	毛国涛, 王杰, 王凯, 王方园, 曹乐言, 张宏森, 宋安东. 水生栖热菌漆酶TaLac的性质分析及对孔雀石绿染料的脱除[J]. 生物技术通报, 2022, 38(4): 261-268.
[4]	田嘉慧, 封佳丽, 卢俊桦, 毛林静, 胡著然, 王莹, 楚杰. 一色齿毛菌漆酶LacT-1的分离纯化与性质研究[J]. 生物技术通报, 2021, 37(8): 186-194.
[5]	陈明雨, 倪烜, 司友斌, 孙凯. 固定化真菌漆酶在环境有机污染修复中的应用研究进展[J]. 生物技术通报, 2021, 37(6): 244-258.
[6]	熊雪, 李鹏, 张贵合, 向准, 陶文广, 周光燕, 和耀威. 不同栽培基质诱导对香菇液体发酵产漆酶活性的影响[J]. 生物技术通报, 2021, 37(12): 50-59.
[7]	王豪, 唐禄鑫, 马鸿飞, 钱坤, 司静, 崔宝凯. 东方栓孔菌漆酶的固定化及其对不同类型染料的脱色作用[J]. 生物技术通报, 2021, 37(11): 142-157.
[8]	陈慧玲, 张青云, 孙凯. 漆酶介导生物体内酚类氧化偶联的基本原理及其在绿色合成中的应用[J]. 生物技术通报, 2020, 36(5): 193-204.
[9]	孙凯, 陈正杰, 汪登洋, 束茹玉, 吴吉, 韦凡. 固定化漆酶去除废水中双酚A[J]. 生物技术通报, 2020, 36(12): 188-198.
[10]	吴怡, 马鸿飞, 曹永佳, 司静, 崔宝凯. 白腐真菌落叶松锈迷孔菌产漆酶液体培养基的优化及其对染料的脱色作用[J]. 生物技术通报, 2020, 36(1): 45-59.
[11]	吴怡, 马鸿飞, 曹永佳, 司静, 崔宝凯. 真菌漆酶的性质、生产、纯化及固定化研究进展[J]. 生物技术通报, 2019, 35(9): 1-10.
[12]	胡楚霄, 雷善钰, 秦艳平, 赵奕锦, 向泉桔. 蒽对3株灵芝菌株漆酶活性及其转录表达水平的影响[J]. 生物技术通报, 2019, 35(9): 112-117.
[13]	韩姝然, 卢磊. 交联酶聚集体的制备及在漆酶固定化中的应用进展[J]. 生物技术通报, 2019, 35(3): 164-170.
[14]	龚睿, 孙凯, 谢道月. 真菌漆酶在绿色化学中的研究进展[J]. 生物技术通报, 2018, 34(4): 24-34.
[15]	钱磊, 张业尼, 陈雪, 刘金福, 张志军, 刘建华. 大球盖菇漆酶的分离纯化及酶学性质研究[J]. 生物技术通报, 2018, 34(4): 127-132.