Engineering Halomonas spp. as A Low-Cost Production Host for Production of Bio-surfactant Protein PhaP.
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.
School of Life Sciences, Tsinghua University, Beijing, China.|Center for Synthetic and Systems Biology, Tsinghua University, Beijing, China.|Tsinghua-Peking Center for Life Sciences, Tsinghua University, Beijing, China.
Lu-Hong
LH
Lan
School of Life Sciences, Tsinghua University, Beijing, China.|Center for Synthetic and Systems Biology, Tsinghua University, Beijing, China.|Tsinghua-Peking Center for Life Sciences, Tsinghua University, Beijing, China.
Han
H
Zhao
School of Life Sciences, Tsinghua University, Beijing, China.
Jin-Chun
JC
Chen
School of Life Sciences, Tsinghua University, Beijing, China.|Center for Synthetic and Systems Biology, Tsinghua University, Beijing, China.|Tsinghua-Peking Center for Life Sciences, Tsinghua University, Beijing, China.|Center for Nano and Micro Mechanics, Tsinghua University, Beijing, China.|MOE Key Lab of Industrial Biocatalysis, Dept Chemical Engineering, Tsinghua University, Beijing, China.
Guo-Qiang
GQ
Chen
D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D019149:Bioreactors; D003365:Costs and Cost Analysis; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D055786:Gene Knockout Techniques; D005818:Genetic Engineering / Q000379:methods*; D020632:Halomonas / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering / Q000191:economics / Q000295:instrumentation / Q000379:methods; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D013501:Surface-Active Agents / Q000737:chemistry / Q000378:metabolism
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; C118828:PHAP protein, Bacteria; D011994:Recombinant Proteins; D013501:Surface-Active Agents
Bio-surfactant; Chromosomal expression; Halomonas; Open Fermentation; PhaP; Synthetic biology