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Cloning of a new allelic variant of aSaccharomyces diastaticus glucoamylase gene and its introduction into industrial yeasts

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Abstract

A new allelic variant of theSTA2 gene, designated asSTA2 K, coding for a secreted glucoamylase, was cloned. Differences were revealed both in the structural gene and in the promoter region, as compared to otherSTA genes. The most peculiar structural features ofSTA2 K are 1. a 1.1-kb natural deletion in its promoter located 189 nucleotides upstream of the translation start codon; and2. an Asn→ Asp single amino acid change within the putative active site of the encoded glucoamylase. Neither the presence of glucose in the medium nor the host cell's mating type constellation affected the expression level ofSTA2 K inS. cerevisiae.

Self-replicating yeast plasmids containingSTA2 K were constructed and used to transform a laboratory yeast strain and various brewing strains. Pilot brewing tests with glucoamylase-secreting transformants of a brewing strain produced superattenuated beers at accelerated fermentation rates.

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Author notes

  1. Keun Kim

    Present address: Institute of Genetic Engineering, Suwon University, Suwon, Korea

  2. Finn Knudsen

    Present address: Beverage Consult International, Inc., Golden, CO

Authors and Affiliations

  1. Biotechnology Center, University of Colorado, 80933-7150, Colorado Springs, CO

    Keun Kim, George Bajszár & James R. Mattoon

  2. Coors Brewing Company, Golden, CO

    Sün Y. Lee & Finn Knudsen

Authors
  1. Keun Kim
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  2. George Bajszár
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  3. Sün Y. Lee
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  4. Finn Knudsen
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  5. James R. Mattoon
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Kim, K., Bajszár, G., Lee, S.Y. et al. Cloning of a new allelic variant of aSaccharomyces diastaticus glucoamylase gene and its introduction into industrial yeasts. Appl Biochem Biotechnol 44, 161–185 (1994). https://doi.org/10.1007/BF02921653

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  • DOI: https://doi.org/10.1007/BF02921653

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