| 来源于链霉菌Streptomyces fradiae var. k11的抗蛋白酶甘露聚糖酶的克隆 |
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| 引用本文： | 邱振华,石鹏君,刘素纯,姚斌.来源于链霉菌Streptomyces fradiae var. k11的抗蛋白酶甘露聚糖酶的克隆[J].中国农业科技导报,2010,12(4):114-120. |
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| 作者姓名： | 邱振华 石鹏君 刘素纯 姚斌 |
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| 作者单位： | (1.湖南农业大学食品科技学院,湖南省食品科学与生物技术重点实验室, 长沙 410128,2.中国农业科学院饲料研究所, 农业部饲料生物技术重点开放实验室, 北京 100081) |
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| 基金项目： | 国家转基因生物新品种培育重大专项，国家肉鸡产业技术体系 |
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| 摘 要： | 通过设计简并引物和TAIL-PCR的方法从Streptomyces fradiae var. k11中克隆得到一个β-l,4-甘露聚糖酶编码基因manS221,其全长1 359 bp,编码452个氨基酸,前端28个氨基酸为预测信号肽。构建表达载体pET-28a-manS221并转化大肠杆菌BL2l（DE3）,特异性表达manS221基因, 重组的甘露聚糖酶通过Ni-NTA亲和层析纯化。酶学性质分析表明,重组甘露聚糖酶最适温度为52℃,最适pH值为6.0,在pH 5.0~9.0的范围内有良好的稳定性,并且对中性蛋白酶具有高抗性。这些优良的性质使得ManS221在食品、饲料和纺织工业生产中具有良好的应用前景。
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| 关 键 词： | 链霉菌Streptomyces fradiae var. k11 甘露聚糖酶 manS221基因 蛋白酶抗性 |
| 收稿时间： | 2010-04-08 |
| 修稿时间： | 2010-05-13 |
Cloning of a Protease-resistant β-mannanase Rooted in Streptomyces fradiae var. k11 |
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| QIU Zheng-hua,SHI Peng-jun,LIU Su Chun,YAO Bin.Cloning of a Protease-resistant β-mannanase Rooted in Streptomyces fradiae var. k11[J].Journal of Agricultural Science and Technology,2010,12(4):114-120. |
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| Authors: | QIU Zheng-hua SHI Peng-jun LIU Su Chun YAO Bin |
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| Institution: | (1.Key Laboratory of Food Science and Biotechnology of Hunan Province, Institute of Food Science and Technology, Hunan Agricultural University, Changsha 410128|2.Key Laboratory of Feed Biotechnology, Ministry of Agriculture|Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China) |
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| Abstract: | The gene manS221 encoding a β-l, 4-mananase was cloned from Streptomyces fradiae var. k11 by degenerate primers and TAIL-PCR technology. manS221 gene contains 1 359 bp, encoding a 452 amino acid peptide consisting of a putative N-terminal signal peptide of 28 residues. The expression vector pET-28a-manS221 was constructed and transformed into Eschetichia coli BL21. The expressed ManS221 was purified by Ni-NTA chromatography. The optimum temperature and pH of the recombinant mannose were 52℃ and 6.0, respectively. The enzyme showed stability over a pH range of 5.0~9.0, and resistance to neutral protease. These favorable properties would make ManS221 as a good candidate in various industrial applications. |
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| Keywords: | Streptomyces fradiae var k11 mannose manS221 gene protease resistance |
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