Highly efficient assimilation of lactose by a metabolically engineered strain ofSaccharomyces cerevisiae

Rubio‐Texeira, Marta; Castrillo, Juan I.; Adam, Ana Cristina; Ugalde, Unai; Polaina, Julio

doi:10.1002/(sici)1097-0061(19980630)14:9<827::aid-yea281>3.0.co;2-n

Cited by 43 publications

(5 citation statements)

References 36 publications

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“…1). This plasmid contains the STA1 gene under control of a galactoseinducible promoter (Latorre-García et al 2005) and a fragment of S. cerevisiae rDNA that directs the integration of the plasmid to the RDN1 locus (Adam et al 1995;Rubio-Texeira et al 1998;Adam et al 1999). Plasmid pSI2 was used to transform strain BY4741 to obtain transformant Y453.…”

Section: Resultsmentioning

confidence: 99%

“…Episomal plasmid pKS2 derives from YEp195 (Wach et al 1994) to which the same 3.5 kb StuI/XbaI DNA insert with the STA1 gene used for pSI2 was added. Yeast DNA was purified by the procedure described by Polaina and Adam (1991) with the modifications described by Rubio-Texeira et al (1998). Further details of plasmid constructions are given in Adam et al (2004) and Latorre-García et al (2005).…”

Section: Plasmid Construction and Dna Techniquesmentioning

confidence: 99%

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Overexpression of the glucoamylase-encoding STA1 gene of Saccharomyces cerevisiae var. diastaticus in laboratory and industrial strains of Saccharomyces

Latorre-García

Adam

Polaina

2008

World J Microbiol Biotechnol

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Production of glucoamylase encoded by the Saccharomyces cerevisiae (var. diastaticus) STA1 gene has been assayed in laboratory S. cerevisiae strains of different ploidy and in different industrial Saccharomyces strains, in which STA1 was expressed under control of an inducible promoter. Highest enzyme activity was achieved with a tetraploid strain constructed by crossing preselected parental strains. Maximal glucoamylase production correlated with heterogeneity in enzyme mass, likely due to incomplete glycosylation, suggesting that the secretionglycosylation process is the limiting step in the production of the STA-encoded glucoamylase by Saccharomyces. Industrial strains showed quite different capacity to produce glucoamylase. High production was achieved with a S. pastorianus brewer's strain. Overall, our results allowed the selection of strains capable of yielding a high level of glucoamylase and suggest specific approaches for further enhancing this capability.

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Section: Resultsmentioning

confidence: 99%

Section: Plasmid Construction and Dna Techniquesmentioning

confidence: 99%

Overexpression of the glucoamylase-encoding STA1 gene of Saccharomyces cerevisiae var. diastaticus in laboratory and industrial strains of Saccharomyces

Latorre-García

Adam

Polaina

2008

World J Microbiol Biotechnol

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“…The engineering of S. cerevisiae for lactose utilization has been addressed over the past 20 years by different strategies [25]. However, most recombinant strains obtained displayed no ideal characteristics (such as slow growth, genetic instability or problems derived from the use of glucose/galactose mixtures) or were ineffective for ethanol production [24,26,27]. There is only one published example of efficient ethanol production with a recombinant S. cerevisiae strain expressing the LAC4 (β-galactosidase) and LAC12 (lactose permease) genes of K. lactis [28].…”

Section: Introductionmentioning

confidence: 99%

A new cold-adapted β-D-galactosidase from the Antarctic Arthrobacter sp. 32c – gene cloning, overexpression, purification and properties

2009

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BackgroundThe development of a new cold-active β-D-galactosidases and microorganisms that efficiently ferment lactose is of high biotechnological interest, particularly for lactose removal in milk and dairy products at low temperatures and for cheese whey bioremediation processes with simultaneous bio-ethanol production.ResultsIn this article, we present a new β-D-galactosidase as a candidate to be applied in the above mentioned biotechnological processes. The gene encoding this β-D-galactosidase has been isolated from the genomic DNA library of Antarctic bacterium Arthrobacter sp. 32c, sequenced, cloned, expressed in Escherichia coli and Pichia pastoris, purified and characterized. 27 mg of β-D-galactosidase was purified from 1 L of culture with the use of an intracellular E. coli expression system. The protein was also produced extracellularly by P. pastoris in high amounts giving approximately 137 mg and 97 mg of purified enzyme from 1 L of P. pastoris culture for the AOX1 and a constitutive system, respectively. The enzyme was purified to electrophoretic homogeneity by using either one step- or a fast two step- procedure including protein precipitation and affinity chromatography. The enzyme was found to be active as a homotrimeric protein consisting of 695 amino acid residues in each monomer. Although, the maximum activity of the enzyme was determined at pH 6.5 and 50°C, 60% of the maximum activity of the enzyme was determined at 25°C and 15% of the maximum activity was detected at 0°C.ConclusionThe properties of Arthrobacter sp. 32cβ-D-galactosidase suggest that this enzyme could be useful for low-cost, industrial conversion of lactose into galactose and glucose in milk products and could be an interesting alternative for the production of ethanol from lactose-based feedstock.

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“…Yeast could also be used for this purpose. However, the considered as one of the best yeast workhorses, Saccharomyces cerevisiae , requires genetic modifications to metabolize lactose (Rubio‐Texeira et al, 1998 ; Sreekrishna & Dickson, 1985 ) and has strong difficulties growing in moderate osmotic media (Norkrans & Kylin, 1969 ). In contrast, other species like some non‐conventional yeasts (e.g., Kluyveromyces lactis , Kluyveromuces marxianus or Candida pseudotripicalis ) have the inherent ability to assimilate and ferment lactose, but none of them can survive in high salinity environments (Karim et al, 2020 ; Sampaio et al, 2020 ; Shen et al, 2019 ).…”

Section: Introductionmentioning

confidence: 99%

Utilization of salt‐rich by‐products from the dairy industry as feedstock for recombinant protein production by Debaryomyces hansenii

Estrada

Navarrete

Møller

et al. 2022

Microbial Biotechnology

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The dairy industry processes vast amounts of milk and generates high amounts of secondary by‐products, which are still rich in nutrients (high Chemical Oxygen Demand (COD) and Biochemical Oxygen Demand (BOD) levels) but contain high concentrations of salt. The current European legislation only allows disposing of these effluents directly into the waterways with previous treatment, which is laborious and expensive. Therefore, as much as possible, these by‐products are reutilized as animal feed material and, if not applicable, used as fertilizers adding phosphorus, potassium, nitrogen, and other nutrients to the soil. Finding biological alternatives to revalue dairy by‐products is of crucial interest in order to improve the utilization of dry dairy matter and reduce the environmental impact of every litre of milk produced. Debaryomyces hansenii is a halotolerant non‐conventional yeast with high potential for this purpose. It presents some beneficial traits – capacity to metabolize a variety of sugars, tolerance to high osmotic environments, resistance to extreme temperatures and pHs – that make this yeast a well‐suited option to grow using complex feedstock, such as industrial waste, instead of the traditional commercial media. In this work, we study for the first time D. hansenii's ability to grow and produce a recombinant protein (YFP) from dairy saline whey by‐products. Cultivations at different scales (1.5, 100 and 500 ml) were performed without neither sterilizing the medium nor using pure water. Our results conclude that D. hansenii is able to perform well and produce YFP in the aforementioned salty substrate. Interestingly, it is able to outcompete other microorganisms present in the waste without altering its cell performance or protein production capacity.

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Highly efficient assimilation of lactose by a metabolically engineered strain ofSaccharomyces cerevisiae

Cited by 43 publications

References 36 publications

Overexpression of the glucoamylase-encoding STA1 gene of Saccharomyces cerevisiae var. diastaticus in laboratory and industrial strains of Saccharomyces

Overexpression of the glucoamylase-encoding STA1 gene of Saccharomyces cerevisiae var. diastaticus in laboratory and industrial strains of Saccharomyces

A new cold-adapted β-D-galactosidase from the Antarctic Arthrobacter sp. 32c – gene cloning, overexpression, purification and properties

Utilization of salt‐rich by‐products from the dairy industry as feedstock for recombinant protein production by Debaryomyces hansenii

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