Functional expression of human serum albumin-tandem thrombopoietin mimetic peptide fusion protein as a novel thrombopoietin analog in Pichia pastoris

被引：4

作者：

Wang, Meizhu ^{[1
]}

Zhi, Dejuan ^{[2
]}

Xian, Jun ^{[1
]}

Ru, Yi ^{[1
]}

Wei, Suzhen ^{[2
]}

Wang, Na ^{[2
]}

Liu, Yiyao ^{[1
]}

Wang, Haiqing ^{[2
]}

Pei, Yuejuan ^{[1
]}

Song, Meijuan ^{[1
]}

Li, Yang ^{[2
]}

Li, Hongyu ^{[1
,2
]}

机构：

[1] Lanzhou Univ, Sch Life Sci, Inst Microbiol, Gansu Key Lab Biomonitoring & Bioremediat Environ, Tianshui South Rd 222, Lanzhou 730000, Peoples R China

[2] Lanzhou Univ, Sch Pharm, Gansu High Throughput Screening & Creat Ctr Hlth, Lanzhou 730000, Peoples R China

来源：

BIOTECHNOLOGY LETTERS | 2016年 / 38卷 / 05期

关键词：

Fusion orientation; Human serum albumin-tandem thrombopoietin mimetic peptide fusion protein; Luciferase reporter assay; Pichia pastoris; Platelet count; Thrombocytopenia; RECEPTOR; AGONIST;

D O I：

10.1007/s10529-016-2049-8

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Objectives To develop a novel thrombopoietin (TPO) analog by fusing the tandem TPO mimetic peptide (TMP-TMP) to human serum albumin (HSA) and performing functional expression of recombinant fusion protein HSA-TMP-TMP. Results After optimizing the fusion orientation in shake-flask culture, HSA-TMP-TMP was expressed at 0.4 g/l in Pichia pastoris grown in a 20 l bioreactor, during which pH was controlled at 5 by addition of NH4OH and citric acid. The fusion protein significantly activated signal transducer and activator of transcription-mediated transcription in TPO receptor-dependent manner, which was demonstrated by a luciferase reporter assay. Following subcutaneous administration, HSA-TMP-TMP effectively stimulated the platelet production in healthy mice in a dose-dependent manner. Conclusion Successful expression of HSA-TMP-TMP fusion protein in P. pastoris was achieved and the recombinant HSA-TMP-TMP is a promising TPO analog.

引用

页码：779 / 785

页数：7

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