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 Mb BioY(MBOVPG45_0349)基因进行系统发育分析、分子对接分析,筛选M.bovis PG45 Mb BioY突变株,比较PG45和PG45ΔMb BioY的生物素敏感性,异源构建Mb BioY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Mb BioY。进一步验证后发现,当M.bovis缺失Mb BioY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Mb BioY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,Mb BioY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。
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 Mb BioY(MBOVPG45_0349)gene was used for phylogenetic analysis and molecular docking analysis;Screening of M.bovis PG45ΔMb BioY mutant;to compare the biotin sensitivity of PG45 and PG45ΔMb BioY the function of M.bovis was further verified by heterologous construction of Mb BioY functional clone.The results showed that a novel biotin membrane transporter Mb BioY 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 Mb BioY loss 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 Mb BioY 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 Mb BioY 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.
作者
陈启伟
权衡
陈胜利
刘东慧
于永峰
李彩玉
宫晓炜
储岳峰
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)
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2024年第1期282-289,共8页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
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)。
关键词
牛支原体
能量耦合因子转运体
生物素
转运
Mycoplasma bovis
energy-coupled factor transporter
biotin
transport
