首页|黑曲霉XZ-3S木聚糖酶基因xynZF-2在毕赤酵母中的高效表达

黑曲霉XZ-3S木聚糖酶基因xynZF-2在毕赤酵母中的高效表达

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将黑曲霉XZ-3S木聚糖酶基因xynZF-2(成熟肽碱基)插入表达载体pPIC9K中,Sal线性化后电击转化毕赤酵母GS115,经MD平板、G418梯度和摇瓶复筛筛选出多克隆阳性转化子.选择28h的种子,每隔24h补加1.5％的甲醇,发酵96h后,比酶活可达13210 U/mg; SDS-PAGE分析表明,该蛋白相对分子质量为23.0ku;重组酶最适作用温度为45℃,最适作用pH为5.0,在温度35～40℃、pH5～7条件下有良好的稳定性,Ca2+对酶的激活作用最明显.
Efficient expression of the xylanase gene xynZF-2 from Aspergillus niger XZ-3S in Pichia pastoris
In this study,a recombinant plasmid pPICgK-xynZF-2was constructed by inserting gene xynZF-2 mature peptide) from Aspergillus niger into Pichia pastoris secretary vector pPICgK.The pPICgK-xynZF-2 linearized by Sal Ⅰ was transformed into Pichia pastoris GS115 by electroporation.The positive recombinant strain was identified by MD medium,G418 and shake bottle selection.The SDS-PAGE analysis showed that the protein molecular weight was about 23.0ku.Under the condition of the vaccination time 28h,1.5％ methanol every 24h added,and the methanol induced time 96h,the enzyme production can be achieved 13210U/mg.The optimal temperature and pH of the enzyme activity was 45℃ and 5.0,respectively.The enzyme activity was stable at the conditions of temperature 35～40℃ and pH 5～7.Ca2+ had an active effect on the enzyme obviously.

Aspergillus nigerxylanasePichia pastoris GS115induction expression

王丹丹、周晨妍、李同彪、张振群、王亚杰、梁真真、张青、路丹荣

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新乡医学院生命科学技术学院,河南新乡453003

新乡医学院三全学院,河南新乡453003

黑曲霉 木聚糖酶 毕赤酵母GS115 诱导表达

河南省教育厅自然科学研究计划项目资助河南省教育厅科学技术研究重点项目河南省科技厅科技攻关项目河南省高等学校青年骨干教师资助计划项目新乡医学院科研项目培育基金

2011A18002613A1808611121022102992011GGJS-1252013ZD113

2015

10.13386/j.issn1002-0306.2015.07.034

食品工业科技
北京一轻研究院

食品工业科技

CSTPCDCSCD北大核心
影响因子：0.842
ISSN：1002-0306
年,卷(期)：2015.36(7)
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