Molecular Biotechnology · 2019 · 11 citations · 72 references
Life SciencesBioengineeringBiotechnologyGene ExpressionMolecular BiotechnologyProtein EngineeringBacterial ToxinsProtein Expression EngineeringProtein PurificationProtein SynthesisRecombinant Protein ProductionToxin ProductionBacterial Protein ProductionBacterial Toxin ProductionCation ExchangeCation-exchange ChromatographyEscherichia Coli Expression SystemsProtein ProductionScalable ProductionToxin-antitoxin Systems
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.
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Green Fluorescent Protein as a Marker for Gene Expression
Martin Chalfie, Yuan Tu, Ghia Euskirchen et al. · Science · 1994 · 6.8K citations