Expression of Recombinant Nematode Anticoagulant Peptide in Pichia Pastoris

China Biotechnology

研究报告

Expression of Recombinant Nematode Anticoagulant Peptide in Pichia Pastoris

Download:

HTML

PDF(0KB) HTML
Export: BibTeX | EndNote (RIS)

Abstract Objective: To acquire recombinant nematode anticoagulant peptide (NAP) with high anticoagulant activity to lay a foundation for the development of novel anticoagulant agent. Methods: Pichia pastoris GS115 strain was transformed with recombinant yeast expression vector pPIC3.5K-rNAP. Expression of rNAP was induced with methanol after the identification of positive strains. NAP expressed in the collected yeast culture supernatant was confirmed with SDS-PAGE and Western blot. The biological activity of the products was validated with PT (prothrombin time), INR (international normalized ratio) and APTT (activated partial thromboplastin time), respectively. Results: The yeast strains stably express NAP were identified. The rNAP was secreted into culture supernatant with a molecular weight of about 10 kD due to glycosylation, which is a little bigger than that predicted (8.7kD). The anticoagulant efficiency of rNAP was confirmed with the in vitro assays. Conclusion: The recombinant nematode anticoagulant peptide with high biological activity was successfully expressed in Pichia pastoris and can be used in the future development of novel anticoagulant agent.

Key words： Pichia pastoris recombinant nematode anticoagulant peptide

Received: 13 January 2006 Published: 25 July 2006

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

Cite this article:

. Expression of Recombinant Nematode Anticoagulant Peptide in Pichia Pastoris. China Biotechnology, 2006, 26(07): 37-41.

URL:

https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2006/V26/I07/37

[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]	CHEN Xin-jie,QIAN Zhi-lan,LIU Qi,ZHAO Qing,ZHANG Yuan-xing,CAI Meng-hao. *Modification of Aromatic Amino Acid Synthetic Pathway in Pichia pastoris* to Produce Cinnamic Acid and ρ-Coumaric Acid**[J]. China Biotechnology, 2021, 41(10): 52-61.

[3]	SHI Peng-cheng, JI Xiao-jun. Advances in Expression of Human Epidermal Growth Factor in Yeast[J]. China Biotechnology, 2021, 41(1): 72-79.

[4]	Yuan TIAN,Yan-ling LI. Biosynthesis of Fusaruside Based on Recombinant Pichia pastoris[J]. China Biotechnology, 2019, 39(7): 8-14.

[5]	Qiang-qiang PENG,Qi LIU,Ming-qiang XU,Yuan-xing ZHANG,Meng-hao CAI. Heterologous Expression of Insulin Precursor in A Newly Engineered Pichia pastoris[J]. China Biotechnology, 2019, 39(7): 48-55.

[6]	Gong CHENG,Si-ming JIAO,Li-shi REN,Cui FENG,Yu-guang DU. *Preparation and Composition Analysis of Chitooligosaccharides with Low Degree of Deacetylation by Hydrolysis of Bacillus subtilis* Chitosanase**[J]. China Biotechnology, 2018, 38(9): 19-26.

[7]	Si-ming JIAO,Gong CHENG,Yu-chen ZHANG,Cui FENG,Li-shi REN,Jian-jun LI,Yu-guang DU. *Expression of Chitinase from Trichoderma reesei* and Analysis the Composition and Structure of its Hydrolysates**[J]. China Biotechnology, 2018, 38(10): 30-37.

[8]	YANG Qing, WANG Bin, WANG Ya-wei, ZHANG Hua-shan, XIONG Hai-rong, ZHANG Li. Comparison of Signal Peptides for Two Hemicellulase Secretory Expression[J]. China Biotechnology, 2017, 37(8): 15-22.

[9]	FENG Xue, GAO Xiang, NIU Chun-qing, LIU Yan. *Construction of Pichia pastoris* Expression Vector of Codon Optimized αB-crystallin Gene and Expression Optimization**[J]. China Biotechnology, 2017, 37(7): 42-47.

[10]	ZHAN Chun-jun, LI Xiang, LIU Guo-qiang, LIU Xiu-xia, YANG Yan-kun, BAI Zhong-hu. *Identification of Glycerol Transporter in Pichia pastoris* and Function Research**[J]. China Biotechnology, 2017, 37(7): 48-55.

[11]	YANG Xu, HUANG Wei-wei, YAO Yu-feng, LIU Cun-bao, SUN Wen-jia, BAI Hong-mei, MA Yan-bing. Impact Factors on the Expression of Recombinant Human Papillomavirus 16 L1 Protein in Pichia pastoris[J]. China Biotechnology, 2017, 37(10): 1-7.

[12]	ZHANG Zhen-yang, YANG Yan-kun, ZHAN Chun-jun, LI Xiang, LIU Xiu-xia, BAI Zhong-hu. Pichia pastoris X-33 ΔGT2 Release the Glycerol Repression on AOX1 and Ef-ficiently Express Heterologous Proteins[J]. China Biotechnology, 2017, 37(1): 38-45.

[13]	KANG Guo-kai, FENG Guo-dong, CAO Kun-lin, CHEN Zheng-jun, GE Xiang-yang. Optimization for High Production Fermentation of Lunasin from Recombinant Pichia pastoris[J]. China Biotechnology, 2016, 36(8): 73-79.

[14]	SHI Hui-lin, SUN Jing-chun, ZHANG Rong-kai, GAO Da-qi, WANG Ze-jian, GUO Mei-jin, ZHOU Li-qin, ZHUANG Ying-ping. Application of the Electronic Nose on the Online Feedback Control of Methanol Concentration during Glucoamylase Fermentation Optimization by Pichia pastoris[J]. China Biotechnology, 2016, 36(3): 68-76.

[15]	LIU Xiao-ming, JIANG Ning, ZHANG Ai-zhong, CAI Peng. Expression of Hybrid Antimicrobial Peptides in Pichia Yeast and Identification of Its Biological Activity[J]. China Biotechnology, 2016, 36(2): 81-89.

Viewed

Full text

Abstract

Cited

Shared

Discussed