Department of Analytical Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387, Kraków, Poland.
Malopolska Centre of Biotechnology, Jagiellonian University, Gronostajowa 7a, 30-387, Kraków, Poland.
Mol Biotechnol. 2019 Oct;61(10):774-782. doi: 10.1007/s12033-019-00203-4.
Lysostaphin is a staphylolytic protein of growing interest from biotechnological and pharmaceutical industry due to its potential use in preventing and combating staphylococcal infections. Here, we describe an optimized method for production of lysostaphin in an inductionless system utilizing constitutive promoter from staphylococcal toxin-antitoxin system PemIK-Sa1. We investigated the influence of ribosome-binding site sequence, Escherichia coli producer strain and growth media on yield and kinetics of recombinant protein production. Lysostaphin was purified in its native active form using one-step cation-exchange chromatography. The system provides a method for cost-efficient and scalable protein production, and can be applied to produce other biotechnologically significant proteins.
溶葡萄球菌酶是一种备受生物技术和制药行业关注的葡萄球菌溶素蛋白，因其在预防和治疗葡萄球菌感染方面的潜在用途而受到关注。在这里，我们描述了一种优化的方法，利用葡萄球菌毒素-抗毒素系统 PemIK-Sa1 的组成型启动子，在无诱导系统中生产溶葡萄球菌酶。我们研究了核糖体结合位点序列、大肠杆菌生产菌株和生长培养基对产量和重组蛋白生产动力学的影响。溶葡萄球菌酶在其天然活性形式下通过一步阳离子交换层析进行纯化。该系统提供了一种经济高效且可扩展的蛋白质生产方法，可用于生产其他具有生物技术意义的蛋白质。
