Dominik L Siebert, Frank Sargent, Ammar Al-Shameri, Volker Sieber
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引用次数: 0
Abstract
Bidirectional [Ni-Fe]-hydrogenases are useful tools for integrating hydrogen into existing chemical processes by utilizing H2 to regenerate expensive cofactors such as NAD(P)H. One enzyme broadly applied to this purpose is the soluble [Ni-Fe]-hydrogenase from Cupriavidus necator (CnSH). However, the homologous production of CnSH suffers from slow growth rates and complex growth medium requirements of the native host. In the present study, we developed a simple approach for the production of CnSH in Escherichia coli based on the coexpression of the maturation factors from C. necator. By optimizing the artificial operons coding for the hydrogenase proteins as well as the maturation factors, we were able to produce CnSH with similar yields and activities compared to the native host. Additionally, we used our system to express three functional novel soluble [Ni-Fe]-hydrogenases, demonstrating its applicability for future enzyme screening and discovery.
期刊介绍：
The journal is particularly interested in studies on the design and synthesis of new genetic circuits and gene products; computational methods in the design of systems; and integrative applied approaches to understanding disease and metabolism.
Topics may include, but are not limited to:
Design and optimization of genetic systems
Genetic circuit design and their principles for their organization into programs
Computational methods to aid the design of genetic systems
Experimental methods to quantify genetic parts, circuits, and metabolic fluxes
Genetic parts libraries: their creation, analysis, and ontological representation
Protein engineering including computational design
Metabolic engineering and cellular manufacturing, including biomass conversion
Natural product access, engineering, and production
Creative and innovative applications of cellular programming
Medical applications, tissue engineering, and the programming of therapeutic cells
Minimal cell design and construction
Genomics and genome replacement strategies
Viral engineering
Automated and robotic assembly platforms for synthetic biology
DNA synthesis methodologies
Metagenomics and synthetic metagenomic analysis
Bioinformatics applied to gene discovery, chemoinformatics, and pathway construction
Gene optimization
Methods for genome-scale measurements of transcription and metabolomics
Systems biology and methods to integrate multiple data sources
in vitro and cell-free synthetic biology and molecular programming
Nucleic acid engineering.
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