Creation of Recombinant Biocontrol Agents by Genetic Programming of Yeast

Acta Naturae

article

Sofiya O. Pipiya¹  Nisso Z. Mirzoeva¹  Margarita N. Baranova¹  Igor E. Eliseev¹  Yuliana A. Mokrushina²  Olga V. Shamova³  Alexander G. Gabibov²  Ivan V. Smirnov⁴  Stanislav S. Terekhov² 

[1]Institute of Bioorganic Chemistry, Russian Academy of Sciences, academicians M.M. Shemyakin and Yu.A. Ovchinnikov, Russian Academy of Sciences

[2]Institute of Bioorganic Chemistry, Russian Academy of Sciences, academicians M.M. Shemyakin and Yu.A. Ovchinnikov, Russian Academy of Sciences, Lomonosov Moscow State University M.V. Lomonosov

[3]Institute of Experimental Medicine

[4]Institute of Bioorganic Chemistry, Russian Academy of Sciences, academicians M.M. Shemyakin and Yu.A. Ovchinnikov, Russian Academy of Sciences, Federal State Budgetary Institution “National Medical Research Center of Endocrinology” of the Ministry of Health of the Russian Federation

关键词: Antimicrobial peptides (AMPs);    yeast Pichia pastoris;    heterologous expression;    protegrin-1 (PG-1);    microfluidic compartmentalization;    emulsion microcultivation;   

DOI  :  10.32607/actanaturae.11878

学科分类：生物技术

来源: Moskovskii Gosudarstvennyi Universitet im.M.V.Lomonosova/M.V.Lomonosov Moscow State University

【 摘 要 】

Bacterial infections caused by antibiotic-resistant pathogens pose an extremely serious and elusive problem in healthcare. The discovery and targeted creation of new antibiotics are today among the most important public health issues. Antibiotics based on antimicrobial peptides (AMPs) are of particular interest due to their genetically encoded nature. A distinct advantage of most AMPs is their direct mechanism of action that is mediated by their membranolytic properties. The low rate of emergence of antibiotic resistance associated with the killing mechanism of action of AMPs attracts heightened attention to this field. Recombinant technologies enable the creation of genetically programmable AMP producers for large-scale generation of recombinant AMPs (rAMPs) or the creation of rAMP-producing biocontrol agents. The methylotrophic yeast Pichia pastoris was genetically modified for the secreted production of rAMP. Constitutive expression of the sequence encoding the mature AMP protegrin-1 provided the yeast strain that effectively inhibits the growth of target gram-positive and gram-negative bacteria. An antimicrobial effect was also observed in the microculture when a yeast rAMP producer and a reporter bacterium were co-encapsulated in droplets of microfluidic double emulsion. The heterologous production of rAMPs opens up new avenues for creating effective biocontrol agents and screening antimicrobial activity using ultrahigh-throughput technologies.

【 授权许可】

Unknown