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Biotechnol J ; 11(12): 1595-1604, 2016 Dec.
Article in English | MEDLINE | ID: mdl-27687610

ABSTRACT

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.


Subject(s)
Bacterial Proteins/metabolism , DNA-Binding Proteins/metabolism , Genetic Engineering/methods , Halomonas/metabolism , Recombinant Proteins/isolation & purification , Recombinant Proteins/metabolism , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Bacterial Proteins/isolation & purification , Bioreactors , Costs and Cost Analysis , DNA-Binding Proteins/chemistry , DNA-Binding Proteins/genetics , DNA-Binding Proteins/isolation & purification , Gene Expression Regulation, Bacterial , Gene Knockout Techniques , Halomonas/genetics , Protein Engineering/economics , Protein Engineering/instrumentation , Protein Engineering/methods , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Surface-Active Agents/chemistry , Surface-Active Agents/metabolism
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