牛支原体能量耦合因子转运蛋白S组件MbBioY摄取外源生物素的功能分析

doi:10.11843/j.issn.0366-6964.2024.01.026

畜牧兽医学报 ›› 2024, Vol. 55 ›› Issue (1): 282-289.doi: 10.11843/j.issn.0366-6964.2024.01.026

牛支原体能量耦合因子转运蛋白S组件MbBioY摄取外源生物素的功能分析

陈启伟, 权衡, 陈胜利, 刘东慧, 于永峰, 李彩玉, 宫晓炜*, 储岳峰*

中国农业科学院兰州兽医研究所/兰州大学动物医学与生物安全学院, 动物疫病防控全国重点实验室甘肃省病原生物学基础学科研究中心, 兰州 730046

收稿日期:2023-03-27 出版日期:2024-01-23 发布日期:2024-01-24
通讯作者: 储岳峰,主要从事动物细菌病的防控技术基础及应用研究,E-mail:chuyuefeng@caas.cn;宫晓炜,主要从事细菌病防控技术及耐药机制研究,E-mail:gongxiaowei@caas.cn
作者简介:陈启伟(1984-),男,甘肃兰州人,副研究员,博士,主要从事细菌病防控技术及耐药机制研究,E-mail:chenqiwei@caas.cn;权衡(1996-),男,陕西宝鸡人,博士生,主要从事细菌病防控技术及耐药机制研究,E-mail:quanheng0517@163.com;陈启伟和权衡为同等贡献作者
基金资助:
中国农业科学院科技创新工程（CAAS-ASTIP-LVRI-04）所级重点任务项目（CAAS-ASTIP-JBGS-20210701）；国家自然科学基金面上项目（32172857）；甘肃省科技重大专项（22ZD6NA001）

Functional Analysis of Biotin Uptake by the Energy-coupled Factor Transporter S Component MbBioY of Mycoplasma bovis

CHEN Qiwei, QUAN Heng, CHEN Shengli, LIU Donghui, YU Yongfeng, LI Caiyu, GONG Xiaowei*, CHU Yuefeng*

Gansu Province Research Center for Basic Disciplines of Pathogen Biology, State Key Laboratory for Animal Disease Control and Prevention, College of Animal Medicine and Biosafety, Lanzhou University/Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China

Received:2023-03-27 Online:2024-01-23 Published:2024-01-24

摘要/Abstract

摘要： 牛支原体（M. bovis）感染与生存依赖于外界微环境提供的各类营养代谢因子。开展M.bovis外源生物素摄取的机制研究，将对M. bovis防控具有极其重要的意义。通过对M. bovis PG45 MbBioY（MBOVPG45_0349）基因进行系统发育分析、分子对接分析，筛选M. bovis PG45 MbBioY突变株，比较PG45 和PG45ΔMbBioY的生物素敏感性，异源构建MbBioY功能性克隆验证其功能。结果表明，PG45染色体上存在一种崭新的生物素膜转运蛋白，全长996 bp，GC含量30.95%，蛋白长度为332个氨基酸，是由7个跨膜结构域组成的能量偶合因子（ECF）转运蛋白 S成分MbBioY。进一步验证后发现，当M.bovis缺失MbBioY会影响其生长，表现出生物素代谢营养缺陷的表型。通过异源敲入生物素代谢营养缺陷大肠杆菌MG1655，表明MbBioY单独存在时，就能够恢复生物素代谢营养缺陷大肠杆菌缺失株的生理功能。初步证明，MbBioY编码的是能量偶合因子（ECF）转运蛋白的S成分具有摄取外源生物素的能力，该研究结果将补充细菌生物素代谢调控的多样性和复杂性，为M.bovis的有效防控提供独特视角。

关键词: 牛支原体, 能量耦合因子转运体, 生物素, 转运

Abstract: M. bovis infection and survival depend on various nutritional and metabolic factors provided 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 examination 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 transmembrane domains. Further verification revealed that MbBioYloss affected M. bovis growth and showed phenotypic biotin metabolism deficiencies. By heterologous knock-in of biotin-metabolizing nutrient-deficient E. coli MG1655, it was shown that MbBioY alone could restore the physiological function of the nutrient-deficient E. coli strain with biotin metabolism. It has been preliminarily 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 diversity and complexity of bacterial biotin metabolism regulation. M. bovis prevention and control will be provided from an innovative perspective.

Key words: Mycoplasma bovis, energy-coupled factor transporter, biotin, transport

中图分类号:

S852.62

陈启伟, 权衡, 陈胜利, 刘东慧, 于永峰, 李彩玉, 宫晓炜, 储岳峰. 牛支原体能量耦合因子转运蛋白S组件MbBioY摄取外源生物素的功能分析[J]. 畜牧兽医学报, 2024, 55(1): 282-289.

CHEN Qiwei, QUAN Heng, CHEN Shengli, LIU Donghui, YU Yongfeng, LI Caiyu, GONG Xiaowei, CHU Yuefeng. Functional Analysis of Biotin Uptake by the Energy-coupled Factor Transporter S Component MbBioY of Mycoplasma bovis[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 282-289.

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牛支原体能量耦合因子转运蛋白S组件MbBioY摄取外源生物素的功能分析

Functional Analysis of Biotin Uptake by the Energy-coupled Factor Transporter S Component MbBioY of Mycoplasma bovis

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