Counterintuitive method improves yields of isotopically labeled proteins expressed in flask-cultured Escherichia coli
Copyright policy )Counterintuitive method improves yields of isotopically labeled proteins expressed in flask-cultured Escherichia coli
Abstract NMR is a powerful tool for the structural and dynamic study of proteins. One of the necessary conditions for the study of these proteins is their isotopic labeling with 13C, 15N and sometimes 2H. One of the most widely used methods to obtain these labeled proteins is heterologous expression of the proteins in E. coli using 13C-D-glucose and 15NH4Cl as the sole nutrient sources. In recent years, the price of 13C-D-glucose has almost tripled, making it essential to develop labeling methods that are as cost effective as possible. In this work, different parameters were studied to achieve the most rational use of 13C-D-glucose, and an optimized method was developed to obtain labeled proteins with high labeling and low 13C-D-glucose consumption. Surprisingly, the optimized method is also simple and does not require monitoring of culture growth.
Carbon Isotopes, Nitrogen Isotopes, Minimal active time, http://metadata.un.org/sdg/3, Article, Recombinant Proteins, Cost reduction, Glucose, 13C labelling, Glucose consumption, Isotope Labeling, Escherichia coli, Protein expression, Nuclear Magnetic Resonance, Biomolecular, Ensure healthy lives and promote well-being for all at all ages
Carbon Isotopes, Nitrogen Isotopes, Minimal active time, http://metadata.un.org/sdg/3, Article, Recombinant Proteins, Cost reduction, Glucose, 13C labelling, Glucose consumption, Isotope Labeling, Escherichia coli, Protein expression, Nuclear Magnetic Resonance, Biomolecular, Ensure healthy lives and promote well-being for all at all ages
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