Fusion expression of bovine lactoferricin in Escherichia coli.
The drug resistance problem has been growing with the utilization of current antibiotics in feed and medical industries. LfcinB, a 25-amino acid antibacterial peptide derived from bovine lactoferrin, is one of potential alternatives of antibiotics. According to the bias of codon utilization of Escherichia coli, a fragment encoding LfcinB has been chemically synthesized, inserted into vector pGEX-4T-2 and expressed in E. coli. The antibacterial peptide was fused with GST with a protease cleavage site located between them. Two constructs with different cleavage sites were made. One construct, pGEX-Th-LfcinB, contains a thrombin cleavage site carried by the vector, and the other, pGEX-Th-Xa-LfcinB, contains a Factor Xa cleavage site which was introduced after the thrombin cleavage site. Fusion protein GST-Th-LfcinB protein was efficiently cleaved by thrombin, yielding recombinant LfcinB showing antibacterial activity. However, fusion protein GEX-Th-Xa-Lfcin B containing Factor Xa recognition site could not be cleaved by Factor Xa at the conditions tried in this study. Successful expression of LfcinB in E. coli provides a possible method to produce LfcinB in large amounts.
Feng
Xing-jun
XJ
Gene Engineering Laboratory, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, PR China.
Wang
Jian-hua
JH
Shan
An-shan
AS
Teng
Da
D
Yang
Ya-lin
YL
Yao
Yi
Y
Yang
Guan-pin
GP
Shao
Yan-chun
YC
Liu
Shuo
S
Zhang
Fan
F
D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000096:biosynthesis / Q000138:chemical synthesis*; D001483:Base Sequence; D002417:Cattle; D003001:Cloning, Molecular / Q000379:methods; D024881:Drug Resistance, Bacterial; D004926:Escherichia coli / Q000235:genetics*; D007781:Lactoferrin / Q000096:biosynthesis* / Q000138:chemical synthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000096:biosynthesis / Q000138:chemical synthesis* / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000138:chemical synthesis / Q000235:genetics*
D000900:Anti-Bacterial Agents; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C082171:lactoferricin B; D007781:Lactoferrin