title: In vitro production and antifungal activity of peptide ABP-dHC-cecropin A.
abstract: The antimicrobial peptide ABP-dHC-cecropin A is a small cationic peptide with potent activity against a wide range of bacterial species. Evidence of antifungal activity has also been suggested; however, testing of this peptide has been limited due to the low expression of cecropin proteins in Escherichia coli. To improve expression of this peptide in E. coli, ABP-dHC-cecropin A was cloned into a pSUMO vector and transformed into E. coli, resulting in the production of a pSUMO-ABP-dHC-cecropin A fusion protein. The soluble form of this protein was then purified by Ni-IDA chromatography, yielding a total of 496-mg protein per liter of fermentation culture. The SUMO-ABP-dHC-cecropin A fusion protein was then cleaved using a SUMO protease and re-purified by Ni-IDA chromatography, yielding a total of 158-mg recombinant ABP-dHC-cecropin A per liter of fermentation culture at a purity of ≥94%, the highest yield reported to date. Antifungal activity assays performed using this purified recombinant peptide revealed strong antifungal activity against both Candida albicans and Neurospora crassa, as well as Rhizopus, Fusarium, Alternaria, and Mucor species. Combined with previous analyses demonstrating strong antibacterial activity against a number of important bacterial pathogens, these results confirm the use of ABP-dHC-cecropin A as a broad-spectrum antimicrobial peptide, with significant therapeutic potential.
authors.affiliation: Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210037, China.
authors.forename: Jiaxin
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210037, China.
authors.forename: Ali
authors.initials: A
authors.lastname: Movahedi
authors.affiliation: Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210037, China.
authors.forename: Junjie
authors.initials: J
authors.lastname: Xu
authors.affiliation: Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210037, China.
authors.forename: Mengyang
authors.initials: M
authors.lastname: Wang
authors.affiliation: Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210037, China.
authors.forename: Xiaolong
authors.initials: X
authors.lastname: Wu
authors.affiliation: Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210037, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Xu
authors.affiliation: Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210037, China.
authors.forename: Tongming
authors.initials: T
authors.lastname: Yin
authors.affiliation: Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210037, China. Electronic address: qzhuge@njfu.edu.cn.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Zhuge
mesh_terms: D000528:Alternaria / Q000187:drug effects; D000935:Antifungal Agents / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology*; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry*; D002176:Candida albicans / Q000187:drug effects; D004926:Escherichia coli / Q000378:metabolism; D008826:Microbial Sensitivity Tests; D009492:Neurospora crassa / Q000187:drug effects; D010455:Peptides / Q000737:chemistry*; D011993:Recombinant Fusion Proteins; D012233:Rhizopus / Q000187:drug effects
chemical_list: D000935:Antifungal Agents; D023181:Antimicrobial Cationic Peptides; D010455:Peptides; D011993:Recombinant Fusion Proteins; C031161:cecropin A
keywords: ABP-dHC-cecropin A; Antibacterial peptide; Antifungal activity; Escherichia coli expression; SUMO