Effect of K202A mutation in the thermostability of Penicillum expansum Lipase

China Biotechnology

2007, Vol. 27

Issue (12): 52-56 DOI:

Effect of K202A mutation in the thermostability of Penicillum expansum Lipase

Download:

HTML

PDF(610KB) HTML
Export: BibTeX | EndNote (RIS)

Abstract

ep8, a thermostabilized mutant of Penicillum expansum lipase obtained in a previous study, contained a single amino acid substitution. To further improve the thermostability of the lipase, the Lys of wild-type (lip07) and mutant (ep8) in 202 were separately substituted by Ala using the Overlap extension PCR technique. The mutant genes (lip07-K202A and ep8-K202A) were subcloned into pAO815, and then transformed into the Pichia pastoris GS115 for extracelluar expression, separately. 15% SDS-PAGE analysis indicated that the molecular mass of PEL-ep8-K202A and PEL-lip07-K202A are both about 28KD, which are just the same with the wild-type lipase. The comparison experiments showed that: The Tm of PEL-ep8-K202A is 41.66℃，2.63℃ higher than that of the wild-type (39.03℃) and 1.21℃ higher than the random mutant(PEL-ep8:40.45℃); the Tm of single mutant (PEL-lip07-K202A) is 37.08℃, 2℃ lower than that of the wild-type lipase.

Key words： Penicillum expansum Lipase site-directed mutagenesis thermostability

Received: 31 August 2007 Published: 25 December 2007

ZTFLH:

Q78

	Service
	E-mail this article
	Add to my bookshelf
	Add to citation manager
	E-mail Alert
	RSS
	Articles by authors

Cite this article:

. Effect of K202A mutation in the thermostability of Penicillum expansum Lipase. China Biotechnology, 2007, 27(12): 52-56.

URL:

https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2007/V27/I12/52

[1]	CHEN Zhong-wei,ZHENG Pu,CHEN Peng-cheng,WU Dan. Screening and Characterization of Thermostable Phytase Mutants[J]. China Biotechnology, 2021, 41(2/3): 30-37.

[2]	MING Yue,ZHAO Zi-tong,WANG Hong-lei,LIANG Zhi-hong. Modification Strategy of Enzyme Thermal Stability Based on Sequence and Structure Analysis[J]. China Biotechnology, 2021, 41(10): 100-108.

[3]	ZHAO Xiao-yan,CHEN Yun-da,ZHANG Ya-qian,WU Xiao-yu,WANG Fei,CHEN Jin-yin. *Site-directed Mutagenesis Improves the Thermostability of Trehalose Synthase TreS II from Myxococcus* sp.V11**[J]. China Biotechnology, 2020, 40(3): 79-87.

[4]	SU Yong-jun,HU Die,HU Bo-chun,LI Chuang,WEN Zheng,ZHANG Chen,WU Min-chen. Improving the Enantioselectivity of an Epoxide Hydrolase towards p-Methylphenyl Glycidyl Ether by Site-directed Mutagenesis[J]. China Biotechnology, 2020, 40(3): 88-95.

[5]	Ting-ting KAN,Xun-cheng ZONG,Yong-jun SU,Ting-ting WANG,Chuang LI,Die HU,Min-chen WU. *Site-directed Mutagenesis of PvEH1 to Improve Its Catalytic Properties towards ortho-Methylphenyl Glycidyl Ether*[J]. China Biotechnology, 2019, 39(6): 9-16.

[6]	Hao-yi MENG,Dan-yang LI,Zheng-yang SUN,Zhao-yong YANG,Zhi-fei ZHANG,Li-jie YUAN. Substrate-binding Site of Ubiquitous Mitochondrial Creatine Kinase from Homo sapiens[J]. China Biotechnology, 2018, 38(5): 24-32.

[7]	LIU Yan-juan, LI Xu-juan, YUAN Hang, LIU Xian, GAO Yan-xiu, GONG Ming, ZOU Zhu-rong. Fusing the Acyl Carrier Protein Enhances the Solubility and Thermostability of the Recombinant Proteins in Escherichia coli[J]. China Biotechnology, 2017, 37(7): 115-123.

[8]	LI Xue-qing, YUAN Feng-jiau, CHENG Jian-qing, DONG Yun-hai, LI Jian-fang, WU Min-chen. Effect of Amino Acid H321 on the Enzymatic Properties of Hybrid β-Mannanase AuMan5A^loop[J]. China Biotechnology, 2017, 37(2): 48-53.

[9]	GUO Chao, WANG Zhi-yan, GAN Yi-ru, LI Dan, DENG Yong, YU Hao-ran, HUANG He. Engineering Thermostability of Bovine Enterokinase by Rational Design Method[J]. China Biotechnology, 2016, 36(8): 46-54.

[10]	WU Qin, HU Die, LI Xue-qing, YUAN Feng-jiao, LI Jian-fang, WU Min-chen. Site-directed Mutagenesis of Y13F to Improve the Thermotolerance of Mesophilic Xylanase from Aspergillus oryzae[J]. China Biotechnology, 2016, 36(12): 36-41.

[11]	XIANG Mian, ZHU Jian-quan, YU Ji-hua, LI Yang-yang, LI Juan-juan, LIU Zu-bi, WANG Wan-jun, LIAO Hai, ZHOU Jia-yu. *The Site-directed Mutation of Key Residues and the Analysis about Inhibitory Activity of Cassia obtusifolia* Trypsin Inhibitor**[J]. China Biotechnology, 2016, 36(10): 15-20.

[12]	LI Yao-yao, BI Jing, WANG Yi-hong, QIN Yun-he, ZHANG Xue-lian. Lysine-322 Acetylation Negatively Regulates Isocitrate Lyase of Mycobacterium tuberculosis[J]. China Biotechnology, 2015, 35(6): 8-13.

[13]	GAO Rui-ping, CHENG Long-bin, LI Zhen-qiu. A Simple and Rapid Single Primer PCR Method for Site-directed Mutagenesis[J]. China Biotechnology, 2015, 35(5): 61-65.

[14]	XIA Ya-mu, LI Chen-chen. Genetic Modification and High Expression of Cyclodextrin Glycosyltransferase[J]. China Biotechnology, 2015, 35(2): 105-110.

[15]	RUAN Jing-jun, YANG Yi, TANG Zi-zhong, CHEN Hui. Study on Tartary Buckwheat Trypsin Inhibitor Activity Sites by Using Site-directed Mutagenesis[J]. China Biotechnology, 2015, 35(12): 30-36.

Viewed

Full text

Abstract

Cited

Shared

Discussed