pBAD＋6xHis＋T7tag＋SerA＋SerB＋SerC
It contains three enzymes of the serine synthesis pathway. Bacteria harboring this plasmidcan be induced by arabinose to synthesize and secrete serine. A SD sequence (RBS) was added to the front of each coding sequence of SerA, SerB and SerC.
pHisx6＋Lpp-OmpA＋FABP
This plasmid expresses a fatty acid binding protein that can be anchoredas a fused protein on the outer membrane. It consists of four basic parts: the Tac promoter, RBS, Lpp-OmpA, and L-FABP. Lpp-OmpA is a sequence encoding a functional peptide that allows surface anchoring of its fused FABP. FABP can bind long chain fatty acids. By transforming this plasmid, the host bacteria can catch long chain fatty acids in liquid and carry them on its outer membrane.
pHisx6＋PelB＋CALB
This plasmid expresses a fusion protein containing a lipase part and can be transported out the of bacterial membrane. The fusion protein can hydrolyze fat to fatty acids. The expression plasmid consists of four basic parts: the Tac promoter, RBS, PelB and CALB. pelB is a sequence encoding a functional signal peptide that helps the CALB fusion protein to be transported out of the membrane. CALB is a lipase that can hydrolyze fat. Being transformed with this plasmid, the host bacteria can secrete lipase out the membrane to hydrolyze fat in the environment to fatty acids.
REFERENCES:
[1]. Matsumiya Y, Wakita D, Kimura A, et al. Isolation and characterization of a lipid-degrading bacterium and its application to lipid-containing wastewater treatment[J]. Journal of Bioscience & Bioengineering, 2007, 103(4):325-330.
[2]. Cammarota M C, Freire D M G. Hydrolytic Enzymes in the Treatment of Wastewater with High Oil and Grease Content[J]. Cheminform, 2006, 37(46):2195.
[3]. Li J, Xia C, Fang X, et al. Identification and characterization of a long-chain fatty acid transporter in the sophorolipid-producing strain Starmerella bombicola[J]. Applied Microbiology & Biotechnology, 2016, 100(16):1-14.
