Purification, kinetic characterization and properties of a novel thermo-tolerant extracellular protease from Kluyveromyces marxianus IFO 0288 with potential biotechnological interest

Foukis, Athanasios; Stergiou, Panagiota‐Yiolanda; Theodorou, Leonidas G.; Papagianni, Maria; Papamichael, Emmanuel M.

doi:10.1016/j.biortech.2012.06.090

Cited by 31 publications

(19 citation statements)

References 28 publications

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“…Decomposition of whey proteins by K. marxianus has been repeatedly studied; the results varied from absent hydrolysis [26], over 20% to 33% [28,29,56], up to 80% hydrolysis [33]. Recent studies confirmed an extracellular serine protease for K. marxianus [57], and Yadav et al [28] proved modification of whey proteins by K. marxianus via electrophoresis. Indigenous proteases in milk [58] and the proteolytic activity of lactobacilli [56] could also contribute some to protein modification during milk processing.…”

Section: Resultsmentioning

confidence: 99%

Efficient growth of Kluyveromyces marxianus biomass used as a biocatalyst in the sustainable production of ethyl acetate

et al. 2015

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Background: Whey is just turning from a waste of milk processing to a renewable raw material in biotechnology for producing single-cell protein, bio-ethanol, or ethyl acetate as an economic alternative. Conversion of whey-borne sugar into ethyl acetate requires yeast biomass as a biocatalyst. A high cell concentration results in a quick ester synthesis, but biomass growth means consumption of sugar at the expense of ester production. Efficient and cost-saving biomass production is thus a practical requirement. Whey is poor in nitrogen and has therefore to be supplemented with a bioavailable N source. Methods: Several aerobic growth tests were performed with Kluyveromyces marxianus DSM 5422 as a potent producer of ethyl acetate in whey-borne media supplemented with various N sources. Preliminary tests were done in shake flasks while detailed studies were performed in a stirred bioreactor.

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Section: Resultsmentioning

confidence: 99%

Efficient growth of Kluyveromyces marxianus biomass used as a biocatalyst in the sustainable production of ethyl acetate

et al. 2015

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“…K. marxianus can also produce eukaryotic proteins since it modifies foreign proteins according to a general eukaryotic scheme. Moreover, K. marxianus has been exploited as a cell factory to obtain valuable enzymes, showing retention of activity in a large temperature interval [11] . …”

Section: Introductionmentioning

confidence: 99%

Thermal adaptability of Kluyveromyces marxianus in recombinant protein production

et al. 2013

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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.

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“…Moreover, K. marxianus has been exploited as a cell factory to produce valuable enzymes, showing retention of the activity in a broad temperature range [120]. The 30X condition showed high expression of INU1 for inulinase, which is useful for the production of recombinant proteins [108,109,121].…”

Section: Transcriptomic Analysismentioning

confidence: 99%

Potential of Thermotolerant Ethanologenic Yeasts Isolated from ASEAN Countries and Their Application in High- Temperature Fermentation

Kosaka¹,

Lertwattanasakul²,

NadchanokRodrussamee³

et al. 2019

Fuel Ethanol Production From Sugarcane

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Thermotolerant ethanologenic yeasts receive attention as alternative bio-ethanol producers to traditionally used yeast, Saccharomyces cerevisiae. Their utilization is expected to provide several benefits for bio-ethanol production due to their characteristics and robustness. They have been isolated from a wide variety of environments in a number of ASEAN countries: Thailand, Vietnam, Laos, and Indonesia. One of these yeasts, Kluyveromyces marxianus has been investigated regarding characteristics. Some strains efficiently utilize xylose, which is a main component of the 2nd generation biomass. In addition, the genetic basis of K. marxianus has been revealed by genomic sequencing and is exploited for further improvement of the strains by thermal adaptation or gene engineering techniques. Moreover, the glucose repression of K. marxianus and its mechanisms has been investigated. Results suggest that K. marxianus is an alternative to S. cerevisiae in next-generation bio-ethanol production industry. Indeed, we have succeeded to apply K. marxianus for bio-ethanol production in a newly developed process, which combines high-temperature fermentation with simultaneous fermentation and distillation under low pressure. This chapter aims to provide valuable information on thermotolerant ethanologenic yeasts and their application, which may direct the economic bioproduction of ethanol and other useful materials in the future.Recently, thermotolerant microorganisms were found among mesophiles with optimum growth temperatures that are 5-10°C higher than those of the typical mesophilic strains Fuel Ethanol Production from Sugarcane 122 Potential of Thermotolerant Ethanologenic Yeasts Isolated from ASEAN Countries… http://dx.doi.org/10.5772/intechopen.79144 123Recently, there have been several reports on ethanol production at high temperatures using P. kudriavzevii (formerly known as I. orientalis). Several P. kudriavzevii strains were reported to grow and produce high levels of ethanol at high temperatures. The strain DMKU 3-ET15 was isolated from traditional fermented pork sausage in Thailand by an enrichment technique in a medium supplemented with 4% ethanol at 40°C. The strain produced 78.6 g/L ethanol from 180 g/L glucose at 40°C [20]. The strain KVMP10 that was isolated from soil located beneath apple trees for ethanol production from orange peel achieved 54 g/L ethanol at 42°C [48]. Strain RZ8-1 that was recently isolated from various samples collected from plant orchards in Thailand produced 33.8 g/L ethanol from 160 g/L glucose at 40°C [49].

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Purification, kinetic characterization and properties of a novel thermo-tolerant extracellular protease from Kluyveromyces marxianus IFO 0288 with potential biotechnological interest

Cited by 31 publications

References 28 publications

Efficient growth of Kluyveromyces marxianus biomass used as a biocatalyst in the sustainable production of ethyl acetate

Efficient growth of Kluyveromyces marxianus biomass used as a biocatalyst in the sustainable production of ethyl acetate

Thermal adaptability of Kluyveromyces marxianus in recombinant protein production

Potential of Thermotolerant Ethanologenic Yeasts Isolated from ASEAN Countries and Their Application in High- Temperature Fermentation

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