| 西兰花硫代葡萄糖苷酶基因BoMyr2在毕赤酵母中的表达 |
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| 引用本文： | 梁锦锋,沈莲清,王向阳,顾青,.西兰花硫代葡萄糖苷酶基因BoMyr2在毕赤酵母中的表达[J].中国食品学报,2010,10(1):67-72. |
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| 作者姓名： | 梁锦锋 沈莲清 王向阳 顾青 |
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| 作者单位： | 浙江工商大学食品和生物工程学院,杭州,310035 |
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| 摘 要： | 以西兰花种子总RNA为模板,通过RT-PCR方法扩增西兰花BoMyr2成熟蛋白基因,将目的片段连接到pMD18-T载体,测序结果显示扩增片段与目的序列一致。通过目的片段的酵母表达载体pPIC9K的构建,获得重组酵母表达载体pPIC9K-BoMyr2,经SacⅠ线性化后电转导入毕赤酵母菌株(Pichia pastoris KM71)。经甲醇诱导表达试验,SDS-PAGE结果显示,表达上清中含有大小约65 kD的蛋白条带,与预期分子质量大小相符。以sinigrin为底物测定表达上清的酶活性,结果显示,通过毕赤酵母KM71表达的培养液具有硫代葡萄糖苷酶活性。硫代葡萄糖苷酶在KM71中的异源表达,为进一步研究与应用提供理论基础。
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| 关 键 词： | 西兰花 硫代葡萄糖苷酶 BoMyr2基因 毕赤酵母 分泌表达 |
Functional Expression of the Myrosinase BoMyr2 from Brassica oleracea var. italica in Yeast Pichia pastoris |
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| Liang Jinfeng,Shen Lianqing,Wang Xiangyang,Gu Qing.Functional Expression of the Myrosinase BoMyr2 from Brassica oleracea var. italica in Yeast Pichia pastoris[J].Journal of Chinese Institute of Food Science and Technology,2010,10(1):67-72. |
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| Authors: | Liang Jinfeng Shen Lianqing Wang Xiangyang Gu Qing |
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| Affiliation: | Liang Jinfeng Shen Lianqing Wang Xiangyang Gu Qing(College of Food Science & Biotechnology Engineering,Zhejiang Industrial , Commercial University,Hangzhou 310035) |
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| Abstract: | Myrosinases are thioglucosidases that hydrolyze the natural plant products glucosinolates which are regarded as precursors of anticancer compounds in the diet.Pichia pastoris expression system was used to express recombinant myrosinase BoMyr2 from Brassica oleracea var.italica.The interested BoMyr2 gene was got by digesting the pMD-BoMyr2 vector using restriction endonuclease,then it was inserted into the secretory pPIC9K Pichia pastoris expression vector and transformed into E.coli.Positive recombinant pla... |
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| Keywords: | Brassica oleracea var italica myrosinase BoMyr2 gene Pichia pastoris secretory expression |
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