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 / Raimondi, Stefano; Elena, Zanni; Amaretti, Alberto; Claudio, Palleschi; Daniela, Uccelletti; Rossi, Maddalena. - In: MICROBIAL CELL FACTORIES. - ISSN 1475-2859. - STAMPA. - 12:(2013), pp. 34-40. [10.1186/1475-2859-12-34]

Thermal adaptability of Kluyveromyces marxianus in recombinant protein production

RAIMONDI, Stefano;AMARETTI, Alberto;ROSSI, Maddalena
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.
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34
40
Thermal adaptability of Kluyveromyces marxianus in recombinant protein production / Raimondi, Stefano; Elena, Zanni; Amaretti, Alberto; Claudio, Palleschi; Daniela, Uccelletti; Rossi, Maddalena. - In: MICROBIAL CELL FACTORIES. - ISSN 1475-2859. - STAMPA. - 12:(2013), pp. 34-40. [10.1186/1475-2859-12-34]
Raimondi, Stefano; Elena, Zanni; Amaretti, Alberto; Claudio, Palleschi; Daniela, Uccelletti; Rossi, Maddalena
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11380/982510
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