Comparison of simple expression procedures in novel expression host Vibrio natriegens and established Escherichia coli system
Autor: | Ján Turňa, Eva Struhárňanská, Ľubica Kormanová, Silvia Rybecká, Jaroslav Blaško, Zdenko Levarski, Lenka Levarská, Stanislav Stuchlík |
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Rok vydání: | 2020 |
Předmět: |
0106 biological sciences
0301 basic medicine Microorganism Bioengineering Vibrio natriegens medicine.disease_cause 01 natural sciences Applied Microbiology and Biotechnology law.invention Metabolic engineering 03 medical and health sciences law 010608 biotechnology Escherichia coli medicine Molecular Biology Vibrio biology Human Growth Hormone Chemistry General Medicine biology.organism_classification Recombinant Proteins 030104 developmental biology Biochemistry Recombinant DNA Fermentation Bacteria Biotechnology |
Zdroj: | Journal of Biotechnology. 321:57-67 |
ISSN: | 0168-1656 |
Popis: | Marine bacterium Vibrio natriegensis a novel host platform for different applications in molecular biology and biotechnology. It has one of the fastest growth rates of any known microorganisms and its extremely short doubling time indicates a high level of proteosynthetic activity. Regarding the necessity of developing new high-level protein expression systems it represents an extremely interesting subject. V. natriegens fulfills many important features for a suitable host including non- pathogenicity, easy scale-up process, potential for using alternative carbon sources (compared to E. coli), growth media and potential for further genetic and metabolic engineering with employment of a wide range of genetic tools. This work compares V. natriegens as an expression host for production of recombinant human growth hormone (hGH), yeast alcohol dehydrogenase (ADH) and archaeal catalase-peroxidase (AfKatG) to E. coliand establishes the basis for future development of this platform. The selected proteins are of different origins, sizes and intended applications. Our results have shown that cultures of V. natriegens using sucrose as a main carbon source can be used for the production of industrially applicable proteins, where it offers higher biomass productions compared to E. coli. In case of human growth hormone production, produced amounts were lower compared to those of E. coli (38 % of total cell protein (TCP) for V. natriegens vs. 58 % of TCP for E. coli, with similar solubility of around 40 % in both cases). In case of yeast alcohol dehydrogenase, V. natriegens produced 26 % of TCP vs. 42 % of TCP in E. coli, but with severely decreased solubility in case of V. natriegens cultures. Finally V. natriegens cultures were able to produce catalase-peroxidase AfKatG at the level of 33 % of TCP compared to 26 % of TCP in E. coli. Obtained results suggest that there are still significant differences in reliability and ease of use between E. coli and V. natriegens, with latter being more susceptible to condition changes and producing inconsistent results. |
Databáze: | OpenAIRE |
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