| 生淀粉结合域SBD及糖化酶基因在毕赤酵母中的融合表达 |
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| 引用本文： | 朱婧,刘海余,王青艳,米慧芝,朱绮霞,廖思明,秦艳,申乃坤,黄日波. 生淀粉结合域SBD及糖化酶基因在毕赤酵母中的融合表达[J]. 广西科学, 2016, 23(1): 7-11. DOI: 10.3969/j.issn.1005-9164.2016.01.002 |
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| 作者姓名： | 朱婧 刘海余 王青艳 米慧芝 朱绮霞 廖思明 秦艳 申乃坤 黄日波 |
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| 作者单位： | 广西科学院国家非粮生物质能源工程技术研究中心,广西南宁,530007 |
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| 基金项目： | 国家自然科学基金项目(31160023)，广西科学研究与技术开发计划项目(桂科合14123001-19)，广西自然科学基金项目(2013GXNSFBA019102)，八桂学者建设工程专项经费和2014年留学人员科技活动项目资助 |
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| 摘 要： | 【目的】研究生淀粉结合域SBD及糖化酶基因在毕赤酵母中的融合表达,提高酶的表达量和水解生淀粉的能力。【方法】利用In-fusionTM PCR克隆技术将淀粉结合域SBD无缝隙插入到黑曲霉糖化酶基因glu的5′端构建融合表达质粒pPIC9K-psg,实现融合酶基因psg在毕赤酵母GS115中的高效表达,并进行酶学性质研究。【结果】融合酶的最适作用条件及热稳定性均与原始酶无明显差别,但反应温度及pH值的范围更为宽泛,在反应温度60~70℃,pH值为4.0~7.0时均较稳定;融合酶PSG降解生淀粉的能力较原始酶PG提高29.6%,比原始菌株提高86.5%。【结论】淀粉结合域SBD的融合提高了糖化酶水解生淀粉的能力。
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| 关 键 词： | 黑曲霉ASP-S21 生淀粉糖化酶 淀粉结合域 In-fusion TM PCR Cloning 融合表达 毕赤酵母GS115 |
| 收稿时间： | 2015-01-27 |
| 修稿时间： | 2015-03-12 |
Fusion Expression of Raw Starch Binding Domain with Glucoamylase in Pichia pastoris |
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| ZHU Jing,LIU Haiyu,WANG Qingyan,MI Huizhi,Zhu Qixi,LIAO Siming,QIN Yan,SHEN Naikun and HUANG Ribo. Fusion Expression of Raw Starch Binding Domain with Glucoamylase in Pichia pastoris[J]. Guangxi Sciences, 2016, 23(1): 7-11. DOI: 10.3969/j.issn.1005-9164.2016.01.002 |
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| Authors: | ZHU Jing LIU Haiyu WANG Qingyan MI Huizhi Zhu Qixi LIAO Siming QIN Yan SHEN Naikun HUANG Ribo |
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| Affiliation: | National Engineering Research Center for Non-food Biorefinery, Guangxi Academy of Sciences, Nanning, Guangxi, 530007, China,National Engineering Research Center for Non-food Biorefinery, Guangxi Academy of Sciences, Nanning, Guangxi, 530007, China,National Engineering Research Center for Non-food Biorefinery, Guangxi Academy of Sciences, Nanning, Guangxi, 530007, China,National Engineering Research Center for Non-food Biorefinery, Guangxi Academy of Sciences, Nanning, Guangxi, 530007, China,National Engineering Research Center for Non-food Biorefinery, Guangxi Academy of Sciences, Nanning, Guangxi, 530007, China,National Engineering Research Center for Non-food Biorefinery, Guangxi Academy of Sciences, Nanning, Guangxi, 530007, China,National Engineering Research Center for Non-food Biorefinery, Guangxi Academy of Sciences, Nanning, Guangxi, 530007, China,National Engineering Research Center for Non-food Biorefinery, Guangxi Academy of Sciences, Nanning, Guangxi, 530007, China and National Engineering Research Center for Non-food Biorefinery, Guangxi Academy of Sciences, Nanning, Guangxi, 530007, China |
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| Abstract: | |
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| Keywords: | Aspergillus niger ASP-S21 raw starch-digesting glucoamylase starch-binding domain(SBD) In-fusion TM PCR Cloning fusion expression Pichia pastoris GS115 |
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