Thermal adaptability of Kluyveromyces marxianus in recombinant protein production

Microbial Cell Factories

Research

Elena Zanni¹  Claudio Palleschi¹  Daniela Uccelletti¹  Maddalena Rossi²  Stefano Raimondi²  Alberto Amaretti² 

[1]Department of Biology and Biotechnology Charles Darwin, University of Rome “La Sapienza”, Piazzale Aldo Moro 5, 00185, Rome, Italy

[2]Department of Life Sciences, University of Modena and Reggio Emilia, Via Campi 183, 41125, Modena, Italy

关键词: Lactis;    Human Serum Albumin;    Specific Growth Rate;    Biomass Yield;    Heterologous Protein;   

DOI  :  10.1186/1475-2859-12-34

 received in 2012-12-21, accepted in 2013-03-26,  发布年份 2013

来源: Springer

【 摘 要 】

BackgroundKluyveromyces marxianus combines the ease of genetic manipulation and fermentation with the ability to efficiently secrete high molecular weight proteins, performing eukaryotic post-translational modifications. It is able to grow efficiently in a wide range of temperatures. The secretion performances were analyzed in the host K. marxianus L3 in the range between 5°C and 40°C by means of 3 different reporter proteins, since temperature appears a key parameter for production and secretion of recombinant proteins.ResultsThe recombinant strains were able to grow up to 40°C and, along the tested temperature interval (5-40°C), the specific growth rates (μ) were generally lower as compared to those of the untransformed strain. Biomass yields were slightly affected by temperature, with the highest values reached at 15°C and 30°C. The secretion of the endogenous β-fructofuranosidase, used as an internal control, was efficient in the range of the tested temperature, as evaluated by assaying the enzyme activity in the culture supernatants. The endogenous β-fructofuranosidase production was temperature dependent, with the highest yield at 30°C. The heterologous proteins HSA, GAA and Sod1p were all successfully produced and secreted between 5°C and 40°C, albeit each one presented a different optimal production temperature (15, 40, 5-30°C for HSA, GAA and Sod1p, respectively).ConclusionsK. marxianus L3 has been identified as a promising and flexible cell factory. In a sole host, the optimization of growth temperatures for the efficient secretion of each individual protein can be carried out over a wide range of temperatures.

【 授权许可】

Unknown   
© Raimondi et al.; licensee BioMed Central Ltd. 2013. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.