Abstract
Genomic DNA and cDNA encoding the β-amylase from the oomycete, Saprolegnia ferax, were cloned into Saccharomyces cerevisiae and analyzed. The Spl. ferax β-amylase gene consisted of a 1350 bp open reading frame, encoding a protein of 450 amino acids with a calculated mass of 49 353 Da, and was not interrupted by any intron. The deduced amino acid sequence of the β-amylase gene had 42% similarity to the β-amylase of Arabidopsis thaliana. The β-amylase gene was expressed in Sacc. cerevisiae and its product was secreted into the culture medium.
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Bai S, Cho NC, Chun SB (1997) Production and characterization of β-amylase from Saprolegnia ferax. Kor. J. Appl. Microbiol. Biotechnol. 25: 115–120.
Bostein D, Falco SC, Stewart SE, Brennan M, Scherer S, Stinchomb DT, Struhl K, Davis RW (1979) Sterile host yeast (SHY). A eukaryotic system of biological containment for recombinant DNA experiments. Gene 8: 17–24.
Chatterjee B, Ghosh A, Das A (1988) Beta-amylolytic activity of Emericella nidulans. Biotechnol. Lett. 10: 143–147.
D'Aguanno VS, Pretorius IS (1994) Optimization of α-amylase and glucoamylase production by recombinant strains of Saccharomyces cerevisiae. Biotechnol. Lett. 16: 727–732.
Gautam SP, Gupta A, Hasija SK, Kekre S (1991) Production and properties of alpha-and beta-amylases from Phaffia rhodozyma and Brettanomyces naardensis. Indian J. Microbiol. 31: 431–434.
Hill J, Ian KA, Donald G, Griffiths DE (1991) DMSO-enhanced whole cell yeast transformation. Nucl. Acids Res. 19: 5791.
Kang HA, Hershey JWB (1994) Effect of initiation factor elF-5A depletion on protein synthesis and proliferation of Saccharomyces cerevisiae. J. Biol. Chem. 269: 3934–3940.
Kim K, Lee JW (1994) Construction of transformed yeast strain secreting both α-amylase and glucoamylase for direct starchfermentation. J. Microbiol. Biotechnol. 4: 7–12.
Kozak M (1984) Compilation and analysis of sequence upstream from the translational start site in eukaryotic mRNAs. Nucl Acids Res. 12: 857–872.
Li XL, Ljungdahl LG (1996) Expression of Aureobasidium pullulans xynA in, and secretion of the xylanase from, Saccharomyces cerevisiae. Appl. Environ. Microbiol. 62: 209–213.
Monroe JD, Salminen MD, Preiss J (1991) Nucleotide sequence of a cDNA clone encoding a β-amylase from Arabidopsis thaliana. Plant Physiol. 97: 1599–1601.
Nanmori T, Nagai M, Shimizu Y, Shinke R, Mikami B (1993) Cloning of the β-amylase gene from Bacillus cereus and characteristics of the primary structure of the enzyme. Appl. Environ. Microbiol. 59: 623–627.
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: A Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. USA 74: 5463–5467.
Shin DJ, Park JC, Lee HB, Chun SB, Bai S (1998) Construction of a secretory expression vector producing an α-amylase of yeast, Schwanniomyces occidentalis in Saccharomyces. J. Microbiol. Biotechnol. 8: 625–630.
von Heijne G (1986) A new method for predicting signal sequence cleavage sites. Nucl. Acids Res. 14: 4683–4690.
Zhu H, Qu F, Zhu LH (1993) Isolation of genomic DNAs from plant, fungi and bacteria using benzyl chloride. Nucl. Acids Res. 21: 5279–5280.
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Kim, HO., Park, JN., Sohn, HJ. et al. Cloning and expression in Saccharomyces cerevisiae of a β-amylase gene from the oomycete Saprolegnia ferax. Biotechnology Letters 22, 1493–1498 (2000). https://doi.org/10.1023/A:1005646013313
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DOI: https://doi.org/10.1023/A:1005646013313