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

共 13 条

[1] Curti E, 2014, HUM VACCINES, V9, P2342
[2] Peptide agonist of the thrombopoietin receptor as potent as the natural cytokine
Cwirla, SE
Balasubramanian, P
Duffin, DJ
Wagstrom, CR
Gates, CM
Singer, SC
Davis, AM
Tansik, RL
Mattheakis, LC
Boytos, CM
Schatz, PJ
Baccanari, DP
Wrighton, NC
Barrett, RW
Dower, WJ
[J]. SCIENCE, 1997, 276 (5319) : 1696 - 1699
[3] High-level expression of a xylanase gene from the thermophilic fungus Paecilomyces thermophila in Pichia pastoris
Fan, Guangsen
Katrolia, Priti
Jia, Huiyong
Yang, Shaoqing
Yan, Qiaojuan
Jiang, Zhengqiang
[J]. BIOTECHNOLOGY LETTERS, 2012, 34 (11) : 2043 - 2048
[4] Human serum albumin: From bench to bedside
Fanali, Gabriella
di Masi, Alessandra
Trezza, Viviana
Marino, Maria
Fasano, Mauro
Ascenzi, Paolo
[J]. MOLECULAR ASPECTS OF MEDICINE, 2012, 33 (03) : 209 - 290
[5] Recovery of major royal jelly protein 1 expressed in Pichia pastoris in aqueous two-phase systems
Ibarra-Herrera, Celeste C.
Torres-Acosta, Mario A.
Mendoza-Ochoa, Gonzalo I.
Aguilar-Yanez, Jose M.
Rito-Palomares, Marco
[J]. JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY, 2014, 89 (07) : 941 - 947
[6] The molecular mechanisms that control thrombopoiesis
Kaushansky, K
[J]. JOURNAL OF CLINICAL INVESTIGATION, 2005, 115 (12) : 3339 - 3347
[7] New thrombopoietic growth factors
Kuter, David J.
[J]. BLOOD, 2007, 109 (11) : 4607 - 4616
[8] The biology of thrombopoietin and thrombopoietin receptor agonists
Kuter, David J.
[J]. INTERNATIONAL JOURNAL OF HEMATOLOGY, 2013, 98 (01) : 10 - 23
[9] Expression of recombinant proteins in Pichia pastoris
Li, Pingzuo
Anumanthan, Anukanth
Gao, Xiu-Gong
Ilangovan, Kuppusamy
Suzara, Vincent V.
Duezguenes, Nejat
Renugopalakrishnan, V.
[J]. APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY, 2007, 142 (02) : 105 - 124
[10] Heterologous protein production using the Pichia pastoris expression system
Macauley-Patrick, S
Fazenda, ML
McNeil, B
Harvey, LM
[J]. YEAST, 2005, 22 (04) : 249 - 270

← 1 2 →