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Stimulation of Bovicin HC5 Production and Selection of Improved Bacteriocin-Producing Streptococcus equinus HC5 Variants

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Abstract

Bovicin HC5 is a peptide that has inhibitory activity against various pathogenic microorganisms and food spoilage bacteria. Aiming to improve the productivity of this bacteriocin, we evaluated several potential factors that could stimulate the synthesis of bovicin HC5 and selected variants of Streptococcus equinus (Streptococcus bovis) HC5 with enhanced bacteriocin production by adaptive laboratory evolution (ALE). The highest production of the bacteriocin (1.5-fold) was observed when Strep. equinus HC5 was cultivated with lactic acid (100 mmol/L). For the ALE experiment, Strep. equinus HC5 cells were subjected to acid-shock (pH 3.0 for 2 h) and maintained in continuous culture for approximately 140 generations (40 days) in media with lactic acid (100 mmol/L) and pH-controlled at 5.5 ± 0.2. An adapted variant was selected showing a distinct phenotype (sedimentation, pigmentation) compared with the parental strain. Bacteriocin production increased 2-fold in this adapted Strep. equinus HC5 variant, which appears to be associated with changes in the cell envelope of the adapted variant and enhanced bacteriocin release into the culture media. In addition, the adapted variant showed higher levels of expression of all bovicin HC5 biosynthetic genes compared with the parental strain during the early and late stages of growth. Results presented here indicate that ALE is a promising strategy for selecting strains of lactic acid bacteria with increased production of bacteriocins.

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Funding

This research was funded by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq; Brasília, Brazil, Grant #432920/2018-8), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES; Brasília, Brazil, Grant #0001), Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG; Belo Horizonte, Brazil), and INCT Ciência Animal.

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  1. Departamento de Microbiologia, Universidade Federal de Viçosa, Vicosa, MG, 36570-900, Brazil

    Sofia Magalhães Moreira & Hilário Cuquetto Mantovani

  2. Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Vicosa, MG, Brazil

    Tiago Antônio de Oliveira Mendes

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  1. Sofia Magalhães Moreira
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Correspondence to Hilário Cuquetto Mantovani.

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All procedures were performed following a protocol approved by the Ethics and Production Animal Care and Use Committee of the Universidade Federal de Viçosa (CEUAP-UFV, Protocol 01/2019).

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Moreira, S.M., de Oliveira Mendes, T.A. & Mantovani, H.C. Stimulation of Bovicin HC5 Production and Selection of Improved Bacteriocin-Producing Streptococcus equinus HC5 Variants. Probiotics & Antimicro. Prot. 13, 899–913 (2021). https://doi.org/10.1007/s12602-020-09703-1

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