Thermal adaptability of Kluyveromyces marxianus in recombinant protein production

Abstract

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.

Developing a broad-range promoter set for metabolic engineering in the thermotolerant yeast Kluyveromyces marxianus

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The Model System Saccharomyces cerevisiae Versus Emerging Non-Model Yeasts for the Production of Biofuels

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Challenging the workhorse: Comparative analysis of eukaryotic micro‐organisms for expressing monoclonal antibodies

“…The methylotrophic yeasts Pichia pastoris (reclassified as Komagataella phaffii and Komagataella pastoris; Cereghino & Cregg, 2007) and Hansenula polymorpha (reclassified as Ogataea polymorpha and Ogataea parapolymorpha; Stockmann et al, 2009) were developed as expression systems with inducible promoters activated by a change in carbon source. Kluyveromyces marxianus has a long history of use in food fermentation and can grow efficiently on inexpensive lactose-based media at a wide variety of temperatures (Raimondi et al, 2013). Arxula adeninivorans (reclassified as Blastobotrys adeninivorans ) is a dimorphic yeast that can grow on a wide variety of carbon and nitrogen sources (Stockmann et al, 2009).…”