Journal
BIOTECHNOLOGY JOURNAL
Volume 11, Issue 12, Pages 1595-1604Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/biot.201600459
Keywords
Bio-surfactant; Chromosomal expression; Halomonas; Open fermentation; PhaP; Synthetic biology
Funding
- State Basic Science Foundation 973 [2012CB725201]
- National Natural Science Foundation of China [31430003, 31270146]
- special Tsinghua President Grant dedicated to this project [2015THZ10]
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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.
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