Background: Kluyveromyces 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. Results: The 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). Conclusions: K. 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.

Thermal adaptability of Kluyveromyces marxianus in recombinant protein production / Stefano, Raimondi; Zanni, Elena; Alberto, Amaretti; Palleschi, Claudio; Uccelletti, Daniela; Maddalena, Rossi. - In: MICROBIAL CELL FACTORIES. - ISSN 1475-2859. - ELETTRONICO. - 12:(2013), p. 34. [10.1186/1475-2859-12-34]

Thermal adaptability of Kluyveromyces marxianus in recombinant protein production

ZANNI, ELENA;PALLESCHI, Claudio;UCCELLETTI, Daniela;
2013

Abstract

Background: Kluyveromyces 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. Results: The 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). Conclusions: K. 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.
2013
Yeast, K.marxianus, recombinant protein production
01 Pubblicazione su rivista::01a Articolo in rivista
Thermal adaptability of Kluyveromyces marxianus in recombinant protein production / Stefano, Raimondi; Zanni, Elena; Alberto, Amaretti; Palleschi, Claudio; Uccelletti, Daniela; Maddalena, Rossi. - In: MICROBIAL CELL FACTORIES. - ISSN 1475-2859. - ELETTRONICO. - 12:(2013), p. 34. [10.1186/1475-2859-12-34]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/523200
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