GIEC OpenIR  > 中国科学院广州能源研究所
Establishment and application of a modified membrane-blot assay for Rhizomucor miehei lipases aimed at improving their methanol tolerance and thermostability
He, Dong1,2,3; Luo, Wen1,5; Wang, Zhiyuan1; Lv, Pengmei1,4; Yuan, Zhenhong1,4; Huang, Shaowei5; Xv, Jingliang1
2017-07-01
发表期刊ENZYME AND MICROBIAL TECHNOLOGY
卷号102页码:35-40
摘要Directed evolution has been proved an effective way to improve the stability of proteins, but high throughput screening assays for directed evolution with simultaneous improvement of two or more properties are still rare. In this study, we aimed to establish a membrane-blot assay for use in the high-throughput screening of Rhizomucor miehei lipases (RMLs). With the assistance of the membrane-blot screening assay, a mutant E47K named G10 that showed improved thermal stability was detected in the first round of error-prone PCR. Using G10 as the parent, two variants G10-11 and G10-20 that showed improved thermal stability and methanol tolerance without loss of activity compared to the wild type RML were obtained. The T-50(60)-value of G10-11 and G10-20 increased by 12 degrees C and 6.5 degrees C, respectively. After incubation for 1 h, the remaining residual activity of G10-11 and G10-20 was 63.45% and 74.33%, respectively, in 50% methanol, and 15.98% and 30.22%, respectively, in 80% methanol. Thus, we successfully developed a membrane-blot assay that could be used for the high-throughput screening of RMLs with improved thermostability and methanol tolerance. Based on our findings, we believe that our newly developed membrane-blot assay will have potential applications in directed evolution in the future.
文章类型Article
关键词Directed Evolution Membrane-blot Methanol Tolerance Thermostability Rhizomucor Miehei Lipases
WOS标题词Science & Technology ; Life Sciences & Biomedicine
资助者Special funds for applied R & D projects of Guangdong Province(2015B020241002) ; Science and Technology Planning Project of Guangdong Province(2016A010104008) ; Special funds for applied R & D projects of Guangdong Province(2015B020241002) ; Science and Technology Planning Project of Guangdong Province(2016A010104008)
DOI10.1016/j.enzmictec.2017.03.010
研究领域[WOS]Biotechnology & Applied Microbiology
关键词[WOS]PICHIA-PASTORIS ; DIRECTED EVOLUTION ; PROTEIN EXPRESSION ; SALT-BRIDGES ; ENZYME ; STABILITY ; RESIDUES ; LACCASE
收录类别SCI
语种英语
资助者Special funds for applied R & D projects of Guangdong Province(2015B020241002) ; Science and Technology Planning Project of Guangdong Province(2016A010104008) ; Special funds for applied R & D projects of Guangdong Province(2015B020241002) ; Science and Technology Planning Project of Guangdong Province(2016A010104008)
WOS类目Biotechnology & Applied Microbiology
WOS记录号WOS:000401688900005
引用统计
文献类型期刊论文
条目标识符http://ir.giec.ac.cn/handle/344007/14122
专题中国科学院广州能源研究所
作者单位1.Chinese Acad Sci, Guangzhou Inst Energy Convers, Key Lab Renewable Energy, Guangzhou 510640, Guangdong, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
3.Chinese Acad Sci, Guangdong Key Lab New & Renewable Energy Res & De, Guangzhou 510640, Guangdong, Peoples R China
4.Collaborat Innovat Ctr Biomass Energy, Zhengzhou 450002, Henan Province, Peoples R China
5.South China Agr Univ, Guangzhou 510642, Guangdong, Peoples R China
推荐引用方式
GB/T 7714
He, Dong,Luo, Wen,Wang, Zhiyuan,et al. Establishment and application of a modified membrane-blot assay for Rhizomucor miehei lipases aimed at improving their methanol tolerance and thermostability[J]. ENZYME AND MICROBIAL TECHNOLOGY,2017,102:35-40.
APA He, Dong.,Luo, Wen.,Wang, Zhiyuan.,Lv, Pengmei.,Yuan, Zhenhong.,...&Xv, Jingliang.(2017).Establishment and application of a modified membrane-blot assay for Rhizomucor miehei lipases aimed at improving their methanol tolerance and thermostability.ENZYME AND MICROBIAL TECHNOLOGY,102,35-40.
MLA He, Dong,et al."Establishment and application of a modified membrane-blot assay for Rhizomucor miehei lipases aimed at improving their methanol tolerance and thermostability".ENZYME AND MICROBIAL TECHNOLOGY 102(2017):35-40.
条目包含的文件
条目无相关文件。
个性服务
推荐该条目
保存到收藏夹
查看访问统计
导出为Endnote文件
谷歌学术
谷歌学术中相似的文章
[He, Dong]的文章
[Luo, Wen]的文章
[Wang, Zhiyuan]的文章
百度学术
百度学术中相似的文章
[He, Dong]的文章
[Luo, Wen]的文章
[Wang, Zhiyuan]的文章
必应学术
必应学术中相似的文章
[He, Dong]的文章
[Luo, Wen]的文章
[Wang, Zhiyuan]的文章
相关权益政策
暂无数据
收藏/分享
所有评论 (0)
暂无评论
 

除非特别说明,本系统中所有内容都受版权保护,并保留所有权利。