Expression of recombinant antibacterial lactoferricin-related peptides from Pichia pastoris expression system.
Four recombinant antimicrobial peptide (rAMP) cDNAs, constructed from two goat lactoferricin-related peptide cDNAs (GLFcin and GLFcin II) with/without (His)(6)-Tag, were cloned into pPICZalphaC and transformed into Pichia pastoris SMD1168H. After methanol induction, these rAMPs were expressed and secreted into broth. They were purified after CM-Sepharose (without His-tg), HisTrap (with His-tg) and Sephadex G-25 chromatographies. The yield of purified rAMP was 0.15 mg/mL of broth. These 4 rAMPs were thermal-stable and with high antibacterial activity against Escherichia coli BCRC 11549, Pseudomonas aeruginosa BCRC 12450, Bacillus cereus BCRC 10603, Staphylococcus aureus BCRC 25923, Propioni bacterium acnes BCRC 10723, and Listera monocytogenes BCRC 14845. The minimum inhibitory concentration (MIC) of rAMPs against these indicators ranged from 4.07 to 16.00 mg/mL.
Department of Cosmetic Science, Providence University, 200 Chung-Chi Road, Taichung 43301, Taiwan.
Gen-Hung
GH
Chen
Wei-Ming
WM
Chen
Guo-Ting
GT
Huang
Yu-Wen
YW
Chen
Shann-Tzong
ST
Jiang
D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D001419:Bacteria / Q000187:drug effects; D015870:Gene Expression; D006041:Goats; D007781:Lactoferrin / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology
D023181:Antimicrobial Cationic Peptides; D011993:Recombinant Fusion Proteins; C082171:lactoferricin B; D007781:Lactoferrin