Functional Analysis of Biotin Uptake by the Energy-coupled Factor Transporter S Component MbBioY of Mycoplasma bovis
M.bovis infection and survival depend on various nutritional and metabolic factors pro-vided by the external microenvironment.The study of the mechanism of exogenous biotin uptake by M.bovis will be of substantial significance to M.bovis prevention and control.An examina-tion of the M.bovis PG45 MbBioY(MBOVPG45_0349)gene was used for phylogenetic analysis and molecular docking analysis;Screening of M.bovis PG45 ΔMbBioY mutant;to compare the biotin sensitivity of PG45 and PG45 ΔMbBioY the function of M.bovis was further verified by heterologous construction of MbBioY functional clone.The results showed that a novel biotin membrane transporter MbBioY was found on the chromosome of PG45.It was 996 bp in length,30.95％GC content,and 332 amino acids in length.The protein consists of seven transmem-brane domains.Further verification revealed that MbBioYloss affected M.bovis growth and showed phenotypic biotin metabolism deficiencies.By heterologous knock-in of biotin-metaboliz-ing nutrient-deficient E.coli MG1655,it was shown that MbBioY alone could restore the physi-ological function of the nutrient-deficient E.coli strain with biotin metabolism.It has been pre-liminarily demonstrated that the S component of the energy coupling factor(ECF)transporter encoded by MbBioY can absorb exogenous biotin.This research result will contribute to the di-versity and complexity of bacterial biotin metabolism regulation.M.bovis prevention and control will be provided from an innovative perspective.
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