High level expression, purification, and characterization of the shrimp antimicrobial peptide, Ch-penaeidin, in Pichia pastoris

Li, Lei; Wang, Jin-Xing; Zhao, Xiao‐Fan; Kang, Cui-Jie; Liu, Ning; Xiang, Jianhai; Li, Fuhua; Sueda, Shinji; Kondo, Hiroki

doi:10.1016/j.pep.2004.09.006

Cited by 78 publications

(40 citation statements)

References 18 publications

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“…A Saccharomyces cerevisiae based expression system proved useful for the production of penaeidin class 3 with respect to the antimicrobial activity spectrum of this class, but non-native glycosylation [46] was a limitation that required troubleshooting [37] and the final product was not C-terminally amidated like native penaeidins. Recently, a penaeidin from Fenneropenaeus chinensis was successfully produced using a methylotrophic yeast expression system [47]. In this case the product included additional residues at the N-terminus which may have affected overall function.…”

Section: Penaeidin Discovery and Evaluation Of Functionmentioning

confidence: 99%

Diversity in penaeidin antimicrobial peptide form and function

Cuthbertson

Deterding

Williams

et al. 2008

Developmental & Comparative Immunology

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Penaeidins are a diverse family of two-domain antimicrobial peptides expressed in shrimp. Variation in penaeidin sequence results in functional diversity, which was discovered using synthetic reproductions of native penaeidins. An isoform of penaeidin class 3 from L. setiferus (Litset Pen3 −4) was synthesized using native ligation and compared directly with the synthetic penaeidin class 4 known to be expressed in the same organism. New antimicrobial activity data is included in this review that emphasizes differences in effectiveness that are apparent from a direct comparison of two classes. A novel approach to intact penaeidin analysis is presented in the form of Fourier Transform Ion-Cyclotron Resonance Mass Spectrometry, which has implications for the identification of individual penaeidin isoforms without chemical modification or enzymatic cleavage. The new information included in this review helps gather the perspective on relevance of penaeidin diversity to antimicrobial function, the use of synthetic peptides as tools to evaluate specific immune functions and the application of high mass resolution, top-down sequencing methods to the intact analysis of individual penaeidin isoforms. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. Keywords NIH Public Access Author ManuscriptDev Comp Immunol. Author manuscript; available in PMC 2009 January 1. Published in final edited form as:Dev Comp Immunol. 2008 ; 32(3): 167-181. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author Manuscript OverviewAntimicrobial peptides (anti-MPs) are a diverse group of innate immune effector molecules that are utilized by multicellular organisms to prevent or combat infection by microbes. AntiMPs have been the subject of extensive review and several categories of anti-MPs have been described based on structural characteristics with certain themes being repeated throughout the living world [1][2][3][4]. Two prominent themes of anti-MP form that are relevant to this particular review include linear, proline-rich peptides, and cysteine-rich peptides that fold into a tertiary structure stabilized by disulfide bonds. These two separate themes provide a lead-in to the peptide family that is the primary focus of this review, the penaeidin family of anti-MPs.Many examples of short anti-MPs found in the literature contain a considerable percentage of Pro residues associated with other specific amino acids, particularly Arg, in repetitive motifs [5][6][7]. Such peptides have been well characterized in insects and mammals. Apidaecin, abaecin, drosocin, formaecin, pyrrhocoricin and lebo...

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Section: Penaeidin Discovery and Evaluation Of Functionmentioning

confidence: 99%

Diversity in penaeidin antimicrobial peptide form and function

Cuthbertson

Deterding

Williams

et al. 2008

Developmental & Comparative Immunology

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“…The translated peptide fowlicidin-2 was successfully released by Kex2, and was shown to have the predicted amino acid sequence and molecular weight. Furthermore, sequencing revealed that there were no additional residues at the N-terminus of the peptide following Kex2 digestion, which was important since the natural N-terminus is critical for the activity of AMPs (25,26).…”

Section: Discussionmentioning

confidence: 99%

Recombinant expression and biological characterization of the antimicrobial peptide fowlicidin-2 in Pichia pastoris

Xing

Tian

Yang

et al. 2016

Experimental and Therapeutic Medicine

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Abstract. Fowlicidins are a group of cathelicidin antimicrobial peptides that were initially identified in chickens. Fowlicidin-2, which is composed of 31 amino acids, is widely expressed in the majority of tissues in chickens and has an important role in innate immunity. In the present study, a recombinant expression system for fowlicidin-2 was successfully constructed using Pichia pastoris X-33 and the expression vector pPICZα-A. Under the optimized fermentation conditions, 85.6 mg fowlicidin-2 with >95% purity was obtained from 1 liter culture medium following purification by ion exchange chromatography and reversed phase high performance liquid chromatography. The recombinant fowlicidin-2 exhibited broad spectrum antimicrobial activity and had a minimum inhibitory concentration ranging from 1 to 4 µM. Furthermore, recombinant fowlicidin-2 exhibited hemolytic activity, promoting 50% human erythrocyte hemolysis in the concentration range of 128-256 µM, and anticancer activity, resulting in the death of 50% of A375 human malignant melanoma cells in the concentration range of 2-4 µM. The results of the present study suggest that recombinant fowlicidin-2 may be a promising candidate for therapeutic applications.

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“…The yeast growth is fast, low cost and as eukaryotes, they have the machinery for post-translational modifications [19]. Recently, many antibacterial peptides have been expressed in Pichia pastoris, including Pisum sativum defensin [6], penaeidin [20], and anti-lipopolysaccharide factor [21].…”

Section: Introductionmentioning

confidence: 99%

Control of apple blue mold by Pichia pastoris recombinant strains expressing cecropin A

Ren

Kong

Wang

et al. 2011

Bioprocess Biosyst Eng

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Recombinant Pichia pastoris yeasts expressing cecropin A (GS115/CEC), was evaluated for the control of the blue mold of apple caused by Penicillium expansum due to cecropin A peptide's effective antimicrobial effects on P. expansum spores by the thiazolyl blue (MTT) assay. Then, the protein concentration was determined and it was expressed at high levels up to 14.2 mg/L in the culture medium. Meanwhile, the population growth was assayed in vivo. The population growth of recombinant strain GS115/CEC was higher than that of non-transformed strain GS115 in red Fuji apples wounds. Recombinant yeast strains GS115/CEC significantly inhibited growth of germinated P. expansum spores in vitro and inhibited decay development caused by P. expansum in apple fruits in vivo when compared with apple fruits inoculated with sterile water or the yeast strain GS115/pPIC (plasmid pPIC9k transformed in GS115). This study demonstrated the potential of expression of the antifungal peptide in yeast for the control of postharvest blue mold infections on pome fruits.

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High level expression, purification, and characterization of the shrimp antimicrobial peptide, Ch-penaeidin, in Pichia pastoris

Cited by 78 publications

References 18 publications

Diversity in penaeidin antimicrobial peptide form and function

Diversity in penaeidin antimicrobial peptide form and function

Recombinant expression and biological characterization of the antimicrobial peptide fowlicidin-2 in Pichia pastoris

Control of apple blue mold by Pichia pastoris recombinant strains expressing cecropin A

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