title: High-level SUMO-mediated fusion expression of ABP-dHC-cecropin A from multiple joined genes in Escherichia coli.
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, evaluation of this peptide has been limited due to the low expression of cecropin proteins in Escherichia coli. To improve the expression level of ABP-dHC-cecropin A in E. coli, tandem repeats of the ABP-dHC-cecropin A gene were constructed and expressed as fusion proteins (SUMO-nABP-dHC-cecropin, n = 1, 2, 3, 4) via pSUMO-nABP-dHC-cecropin A vectors (n = 1, 2, 3, 4). Comparison of the expression levels of soluble SUMO-nABP-dHC-cecropin A fusion proteins (n = 1, 2, 3, 4) suggested that BL21 (DE3)/pSUMO-3ABP-dHC-cecropin A is an ideal recombinant strain for ABP-dHC-cecropin A production. Under the selected conditions of cultivation and isopropylthiogalactoside (IPTG) induction, the expression level of ABP-dHC-cecropin A was as high as 65 mg/L, with ∼21.3% of the fusion protein in soluble form. By large-scale fermentation, protein production reached nearly 300 mg/L, which is the highest yield of ABP-dHC-cecropin A reported to date. In antibacterial experiments, the efficacy was approximately the same as that of synthetic ABP-dHC-cecropin A. This method provides a novel and effective means of producing large amounts of ABP-dHC-cecropin A.
authors.affiliation: Co-Innovation Center for Sustainable Forestry in Southern China, Key Laboratory of Forest Genetics & Biotechnology, Ministry of Education, 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, Key Laboratory of Forest Genetics & Biotechnology, Ministry of Education, 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, Key Laboratory of Forest Genetics & Biotechnology, Ministry of Education, Nanjing Forestry University, Nanjing 210037, China.
authors.forename: Zhiheng
authors.initials: Z
authors.lastname: Wei
authors.affiliation: Co-Innovation Center for Sustainable Forestry in Southern China, Key Laboratory of Forest Genetics & Biotechnology, Ministry of Education, Nanjing Forestry University, Nanjing 210037, China.
authors.forename: Ming
authors.initials: M
authors.lastname: Sang
authors.affiliation: Co-Innovation Center for Sustainable Forestry in Southern China, Key Laboratory of Forest Genetics & Biotechnology, Ministry of Education, 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, Key Laboratory of Forest Genetics & Biotechnology, Ministry of Education, 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, Key Laboratory of Forest Genetics & Biotechnology, Ministry of Education, Nanjing Forestry University, Nanjing 210037, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Wei
authors.affiliation: Co-Innovation Center for Sustainable Forestry in Southern China, Key Laboratory of Forest Genetics & Biotechnology, Ministry of Education, Nanjing Forestry University, Nanjing 210037, China.
authors.forename: Huixin
authors.initials: H
authors.lastname: Pan
authors.affiliation: Co-Innovation Center for Sustainable Forestry in Southern China, Key Laboratory of Forest Genetics & Biotechnology, Ministry of Education, 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, Key Laboratory of Forest Genetics & Biotechnology, Ministry of Education, Nanjing Forestry University, Nanjing 210037, China. Electronic address: qzhuge@njfu.edu.cn.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Zhuge
mesh_terms: D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D025842:SUMO-1 Protein / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D023181:Antimicrobial Cationic Peptides; D011993:Recombinant Fusion Proteins; D025842:SUMO-1 Protein; C031161:cecropin A
keywords: ABP-dHC-cecropin A; Antimicrobial peptide; Hydroxylamine hydrochloride; Large-scale fermentation; Multimers; SUMO