Abstract
Although Escherichia coli and Bacillus subtilis are the most prominent bacterial hosts for recombinant protein production by far, additional species are being explored as alternatives for production of difficult-to-express proteins. In particular, for thermostable proteins, there is a need for hosts able to properly synthesize, fold, and excrete these in high yields, and thermophilic Bacillaceae represent one potentially interesting group of microorganisms for such purposes. A number of thermophilic Bacillaceae including B. methanolicus, B. coagulans, B. smithii, B. licheniformis, Geobacillus thermoglucosidasius, G. kaustophilus, and G. stearothermophilus are investigated concerning physiology, genomics, genetic tools, and technologies, altogether paving the way for their utilization as hosts for recombinant production of thermostable and other difficult-to-express proteins. Moreover, recent successful deployments of CRISPR/Cas9 in several of these species have accelerated the progress in their metabolic engineering, which should increase their attractiveness for future industrial-scale production of proteins. This review describes the biology of thermophilic Bacillaceae and in particular focuses on genetic tools and methods enabling use of these organisms as hosts for recombinant protein production.
Highlights
Bacteria are widely used in science and industry as cell factories for production of recombinant proteins
B. subtilis, on the other hand, has become an industrial workhorse for recombinant protein production due to an easy cultivation, the products of its metabolism being generally recognized as safe (GRAS), ease of genetic manipulation, well-characterized expression systems, absence of significant codon bias, and exceptional ability to secrete heterologous proteins allowing cost-effective downstream processing
Thermophilic Bacillaceae are currently not used for commercial production of heterologous proteins; their favorable physiological properties, together with current development of genetic toolboxes as presented in this review, should indicate the feasibility of these organisms as promising hosts for recombinant production in the future
Summary
Bacteria are widely used in science and industry as cell factories for production of recombinant proteins. E. coli is in many cases the most used host due to several advantages, including an extensively developed genetic tool box, well-known genetics and physiology, low-cost media, and rapid protein production in a short fermentation period [1] This species exhibits some limitations as the heterologous proteins are typically expressed intracellularly, which results in problems with formation of inclusion bodies and incorrect protein folding. Thermophilic Bacillaceae are currently not used for commercial production of heterologous proteins; their favorable physiological properties, together with current development of genetic toolboxes as presented in this review, should indicate the feasibility of these organisms as promising hosts for recombinant production in the future. Genome scale model: (M10EXG) [69] and (DL33) [70] Ethanol, 1 L (NCIMB11955) [31]
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