Engineering Halomonas spp. as a low-cost production host for production of bio-surfactant protein PhaP

被引:45
作者
Lan, Lu-Hong [1 ,2 ,3 ]
Zhao, Han [1 ,2 ,3 ]
Chen, Jin-Chun [1 ]
Chen, Guo-Qiang [1 ,2 ,3 ,4 ,5 ]
机构
[1] Tsinghua Univ, Sch Life Sci, Beijing 100084, Peoples R China
[2] Tsinghua Univ, Ctr Synthet & Syst Biol, Beijing, Peoples R China
[3] Tsinghua Univ, Tsinghua Peking Ctr Life Sci, Beijing, Peoples R China
[4] Tsinghua Univ, Ctr Nano & Micro Mech, Beijing, Peoples R China
[5] Tsinghua Univ, Dept Chem Engn, MOE Key Lab Ind Biocatalysis, Beijing, Peoples R China
基金
中国国家自然科学基金;
关键词
Bio-surfactant; Chromosomal expression; Halomonas; Open fermentation; PhaP; Synthetic biology; ESCHERICHIA-COLI; RECOMBINANT PROTEINS; EXPRESSION; GENE; PURIFICATION; SYSTEM; POLYHYDROXYBUTYRATE; GRANULES; PHASIN; TD01;
D O I
10.1002/biot.201600459
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Halomonas spp. have been studied as a low cost production host for producing bulk materials such as polyhydroxyalkanoates (PHA) bioplastics, since they are able to grow at high pH and high NaCl concentration under unsterile and continuous conditions without microbial contamination. In this paper, Halomonas strain TD is used as a host to produce a protein named PHA phasin or PhaP which has a potential to be developed into a bio-surfactant. Four Halomonas TD expression strains are constructed based on a strong T7-family expression system. Of these, the strain with phaC deletion and chromosomal expression system resulted in the highest production of PhaP in soluble form, reaching 19% of total cellular soluble proteins and with a yield of 1.86 g/L in an open fed-batch fermentation process. A simple "heat lysis and salt precipitation" method is applied to allow rapid PhaP purification from a mixture of cellular proteins with a PhaP recovery rate of 63%. It clearly demonstrated that Halomonas TD could be used for high yield expression of a bio-surfactant protein PhaP for industrial application in an economical way.
引用
收藏
页码:1595 / 1604
页数:10
相关论文
共 35 条
[1]   THE STABILIZATION OF PROTEINS BY OSMOLYTES [J].
ARAKAWA, T ;
TIMASHEFF, SN .
BIOPHYSICAL JOURNAL, 1985, 47 (03) :411-414
[2]   DEVELOPMENT OF A GENE REPORTER SYSTEM IN MODERATELY HALOPHILIC BACTERIA BY EMPLOYING THE ICE NUCLEATION GENE OF PSEUDOMONAS-SYRINGAE [J].
ARVANITIS, N ;
VARGAS, C ;
TEGOS, G ;
PERYSINAKIS, A ;
NIETO, JJ ;
VENTOSA, A ;
DRAINAS, C .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1995, 61 (11) :3821-3825
[3]   Comparative genomics study of polyhydroxyalkanoates (PHA) and ectoine relevant genes from Halomonas sp TD01 revealed extensive horizontal gene transfer events and co-evolutionary relationships [J].
Cai, Lei ;
Tan, Dan ;
Aibaidula, Gulsimay ;
Dong, Xin-Ran ;
Chen, Jin-Chun ;
Tian, Wei-Dong ;
Chen, Guo-Qiang .
MICROBIAL CELL FACTORIES, 2011, 10
[4]   Replicon-Free and Markerless Methods for Genomic Insertion of DNAs in Phage Attachment Sites and Controlled Expression of Chromosomal Genes in Escherichia coli [J].
Chiang, Chung-Jen ;
Chen, Po Ting ;
Chao, Yun-Peng .
BIOTECHNOLOGY AND BIOENGINEERING, 2008, 101 (05) :985-995
[5]   SURFACE-ACTIVE AGENTS FROM 2 BACILLUS SPECIES [J].
COOPER, DG ;
GOLDENBERG, BG .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1987, 53 (02) :224-229
[6]   The improvement of fibroblast growth on hydrophobic biopolyesters by coating with polyhydroxyalkanoate granule binding protein PhaP fused with cell adhesion motif RGD [J].
Dong, Ying ;
Li, Ping ;
Chen, Chong-bo ;
Wang, Zhi-hui ;
Ma, Ping ;
Chen, Guo-Qiang .
BIOMATERIALS, 2010, 31 (34) :8921-8930
[7]   Intensified expression and purification of a recombinant biosurfactant protein [J].
Dwyer, Mirjana Dimitrijev ;
Brech, Michael ;
Yu, Lei ;
Middelberg, Anton P. J. .
CHEMICAL ENGINEERING SCIENCE, 2014, 105 :12-21
[8]  
Friehs K, 2004, ADV BIOCHEM ENG BIOT, V86, P47
[9]   Cloning and expression of α-amylase from the hyperthermophilic archaeon Pyrococcus woesei in the moderately halophilic bacterium Halomonas elongata [J].
Frillingos, S ;
Linden, A ;
Niehaus, F ;
Vargas, C ;
Nieto, JJ ;
Ventosa, A ;
Antranikian, G ;
Drainas, C .
JOURNAL OF APPLIED MICROBIOLOGY, 2000, 88 (03) :495-503
[10]   Development of Halomonas TD01 as a host for open production of chemicals [J].
Fu, Xiao-Zhi ;
Tan, Dan ;
Aibaidula, Gulsimay ;
Wu, Qiong ;
Chen, Jin-Chun ;
Chen, Guo-Qiang .
METABOLIC ENGINEERING, 2014, 23 :78-91