Laboratorio de Microbiología Molecular, Departamento de Cienciay Tecnología, Universidad Nacional de Quilmes, Bernal, Argentina.
Biotechnol Lett. 2012 Jul;34(7):1275-81. doi: 10.1007/s10529-012-0887-6. Epub 2012 Mar 4.
Prokaryotes commonly present outer cell wall structures composed of a crystalline array of proteinaceous subunits, known as surface layers (S-layers). The ORF encoding the S-layer protein (SlpA) of Lactobacillus brevis was cloned into Lactococcus lactis under the transcriptional control of the xylose-inducible expression system (XIES). SlpA was secreted into the extracellular medium, as determined by immunoblotting, and assays on the kinetics of SlpA production revealed that repression of the system with glucose did not require the depletion of xylose from the medium that allows transitory ORF expression. The successful use of XIES to express S-layer proteins in the versatile and generally recognized as safe species L. lactis opens new possibilities for an efficient production and isolation of SlpA S-layer protein for its various applications in biotechnology and importantly as an antigen-carrying vehicle.
原核生物通常具有由蛋白质亚基组成的结晶状细胞外细胞壁结构,称为表面层 (S-layer)。短小乳杆菌的 S-层蛋白 (SlpA) 的开放阅读框 (ORF) 被克隆到乳球菌 lactis 中,受木糖诱导表达系统 (XIES) 的转录控制。通过免疫印迹测定,SlpA 被分泌到细胞外培养基中,并且对 SlpA 产生动力学的测定表明,葡萄糖对系统的抑制并不需要从允许瞬时 ORF 表达的培养基中耗尽木糖。XIES 成功地在多功能且被普遍认为安全的乳球菌 lactis 中表达 S-层蛋白,为高效生产和分离 SlpA S-层蛋白及其在生物技术中的各种应用(重要的是作为抗原载体)开辟了新的可能性。