Magnetic nanoparticles mediate the transformation of antimicrobial peptides HeM into Chlorella ellipsoidea
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Magnetic nanoparticles mediate the transformation of antimicrobial peptides HeM into Chlorella ellipsoidea Sushan Cao 1,2 & Jing Xue 1 & Xuqing Chen 1 & Xianhui An 2 & Xiuhai Zhang 1 Received: 28 November 2019 / Revised and accepted: 5 March 2020 # Springer Nature B.V. 2020
Abstract Antimicrobial peptides are short peptides with broad-spectrum antimicrobial properties. Heliomicin, a peptide from Heliothis virescens, was re-engineered to have both antifungal and bacterial activities and renamed as HeM. Chlorella is a perfect bioreactor candidate due to fast growth, high photosynthetic efficiency, and easy large-scale culture. This study describes the successful transformation of HeM into Chlorella ellipsoidea F962. A binary expression vector pGcp4 was constructed that carried the antimicrobial peptide gene HeM and the gene encoding neomycin phosphotransferase II (NPT II). The pGcp4 was introduced into C. ellipsoidea F962 by magnetic nanoparticle-mediated transformation method. The positive transformants were obtained by G418 resistance screening. Molecular and genetic analyses on transformants revealed that the antimicrobial peptide gene and the NPT II gene had successfully entered C. ellipsoidea and expressed antimicrobial activity in transgenic cells. This method can provide a new way to produce transgenic Chlorella as well as explore wide utilization of Chlorella through magnetic transformation methods to mediate the production of antimicrobial peptides by C. ellipsoidea. Keywords Antimicrobial peptides . Chlorella ellipsoidea . Transformation . Magnetic nanoparticles
Introduction As a short peptide substance derived from natural organisms, antimicrobial peptides are widely found in animals, plants, and microorganisms. More than a thousand of antimicrobial peptides have been discovered (Wang et al. 2015). After intensive research, it was found that a variety of antimicrobial peptides generally have the characteristics of short peptide chain and good thermal stability (Li et al. 2012). Antimicrobial peptides have different metabolism and action mechanisms than traditional antibiotics and exhibit broad-spectrum antibacterial properties against bacteria, fungus, and viruses, and hardly produce drug resistance (Epan and Vogel 1999). As the most promising active substance currently expected to replace antibiotics, they have the
* Xiuhai Zhang [email protected] 1
Beijing Key Laboratory of Agricultural Genetic Resources and Biotechnology, Beijing Agro-Biotechnology Research Center, Beijing Academy of Agriculture and Forestry Sciences, Beijing 10097, China
2
Jiangsu Key Laboratory of Marine Bioresources and Eco-environment Lianyungang, College of Marine Life and Fisheries, Jiangsu Ocean University, Lianyungang 222005, Jiangsu, China
characteristics that antibiotics do not have. The structure, mechanism of action, and safety of various antimicrobial peptides have been studied (Hong et al. 2016; Gagnon et al. 2016; Kocourková et al. 2017; Jiang et al. 2018; Santos et al. 2018). More and more researchers have
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