Display of adenoregulin with a novel Pichia pastoris cell surface display system.
Two Pichia pastoris cell surface display vectors were constructed. The vectors consisted of the flocculation functional domain of Flo1p with its own secretion signal sequence or the alpha-factor secretion signal sequence, a polyhistidine (6xHis) tag for detection, an enterokinase recognition site, and the insertion sites for target proteins. Adenoregulin (ADR) is a 33-amino-acid antimicrobial peptide isolated from Phyllomedusa bicolor skin. The ADR was expressed and displayed on the Pichia pastoris KM71 cell surface with the system reported. The displayed recombinant ADR fusion protein was detected by fluorescence microscopy and confocal laser scanning microscopy (CLSM). The antimicrobial activity of the recombinant adenoregulin was detected after proteolytic cleavage of the fusion protein on cell surface. The validity of the Pichia pastoris cell surface display vectors was proved by the displayed ADR.
Ren
Ren
R
State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai 200237, PR China.
Jiang
Zhengbing
Z
Liu
Meiyun
M
Tao
Xinyi
X
Ma
Yushu
Y
Wei
Dongzhi
D
D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D001001:Anura / Q000235:genetics / Q000378:metabolism; D001709:Biotechnology; D002462:Cell Membrane / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005800:Genes, Fungal; D005822:Genetic Vectors; D066298:In Vitro Techniques; D037241:Mannose-Binding Lectins / Q000235:genetics; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009479:Neuropeptides / Q000235:genetics* / Q000378:metabolism* / Q000494:pharmacology; D010843:Pichia / Q000235:genetics* / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics
D023181:Antimicrobial Cationic Peptides; C087578:FLO1 protein, S cerevisiae; D037241:Mannose-Binding Lectins; D009479:Neuropeptides; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; C078563:adrenoregulin