Molecular cloning of the glucoamylase gene of Aspergillus shirousami and its expression in Aspergillus oryzae.

Shibuya, Ichiro; Gomi, Katsuya; Iimura, Yuzuru; Takahashi, Koichi; Tamura, Gakuzo; Hara, Shodo

doi:10.1271/bbb1961.54.1905

Cited by 31 publications

(11 citation statements)

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“…Subsequently, the 1.4-kb DNA fragment containing the coding and terminator region of cgsG was inserted immediately downstream of the actA promoter in pKW13000 to yield pKW12203, a hygromycin resistance-conferring vector capable of overexpressing cgsG (Figure 1b). Transformation was carried out as described elsewhere 6 with the following modification: the recipient C. globosum strain was initially cultivated in MYG medium containing 0.8 M sorbitol and 200 mg ml À1 hygromycin. The plasmid pKW12203 was linearized by XbaI and transformed into C. globosum.…”

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confidence: 99%

Overexpressing transcriptional regulator in Chaetomium globosum activates a silent biosynthetic pathway: evaluation of shanorellin biosynthesis

et al. 2012

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confidence: 99%

Overexpressing transcriptional regulator in Chaetomium globosum activates a silent biosynthetic pathway: evaluation of shanorellin biosynthesis

et al. 2012

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“…The amino acid sequence of GA I was similar to glucoamylase from A. shirousamii (Shibuya et al, 1990) and A. niger (Boel et al, 1984) with arrange of homology from 95 to 97%. It is also known that the amino acid sequences of the four domains that form the active center is highly conserved at glucoamylases.…”

Section: Resultsmentioning

confidence: 88%

“…N-terminal amino acid sequence of GA I was ATLDSWLSNEATVARTA. This sequence showed complete homology with glucoamylase from A. awamori (Nunberg et al, 1984), A. niger (Boel et al, 1984) and A. shirousamii (Shibuya et al, 1990). However, only 11 residues were homologous with that of glucoamylase from A. oryzae (Hata et al, 1991).…”

Section: Resultsmentioning

confidence: 90%

Molecular Cloning and Determination of the Nucleotide Sequence of Raw Starch Digesting α-Amylase from Aspergillus awamori KT-11

Matsubara¹,

Ammar²,

Anindyawati³

et al. 2004

BMB Reports

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Complementary DNAs encoding α-amylases (Amyl I, Amyl III) and glucoamylase (GA I) were cloned from Aspergillus awamori KT-11 and their nucleotide sequences were determined. The sequence of Amyl III that was a raw starch digesting α-amylase was found to consist of a 1,902 bp open reading frame encoding 634 amino acids. The signal peptide of the enzyme was composed of 21 amino acids. On the other hand, the sequence of Amyl I, which cannot act on raw starch, consisted of a 1,500 bp ORF encoding 499 amino acids. The signal peptide of the enzyme was composed of 21 amino acids. The sequence of GA I consisted of a 1,920 bp ORF that encoded 639 amino acids. The signal peptide was composed of 24 amino acids. The amino acid sequence of Amyl III from the N-terminus to the amino acid number 499 showed 63.3% homology with Amyl I. However, the amino acid sequence from the amino acid number 501 to C-terminus, including the rawstarch-affinity site and the TS region rich in threonine and serine, showed 66.9% homology with GA I.

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“…3), and four putative glycosylation sites were at amino acid positions 93 (Asn-Pro-Ser), 170 (Asn-Gln-Thr), 181 (Asn-Gly-Ser), and 394 (Asn-GlySer), in the glucoamylase. 10 ) Some of these sites may be glycosylated in the ex-amylase and glucoamylase secreted from the S. cerevisiae transfomants.…”

Section: Discussionmentioning

confidence: 99%

Cloning of the α-Amylase cDNA ofAspergillus shirousamiiand Its Expression inSaccharomyces cerevisiae

Shibuya

Tamura

Ishikawa

et al. 1992

Bioscience, Biotechnology, and Biochemistry

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~-Amylase cDNA was cloned and sequenced from Aspergillus shirousamii RIB2504. The putative protein deduced from the cDNA open reading frame (ORF) consisted of 499 amino acids with a molecular weight of 55,000. The amino acid sequence was identical to that of the ORF of the Taka-amylase A gene of Aspergillus oryzae, while the nucleotide sequence was different at two and six positions in the cDNA ORF and 3' non-coding regions, respectively, so far determined. The ~-amylase cDNA was expressed in Saccharomyces cerevisiae under the control of the yeast ADHl promoter using a YEp-type plasmid, pYcDEl. The cDNA of glucoamylase, which was previously cloned from the same organism, was also expressed under the same conditions. Consequently, active ~-amylase and glucoamylase were efficiently secreted into the culture medium. The amino acid sequence of the N-terminal regions of these enzymes purified from the yeast culture medium confirmed that the signal sequences of these enzymes were cleaved off at the same positions as those of the native enzymes of A. shirousamii.

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Molecular cloning of the glucoamylase gene of Aspergillus shirousami and its expression in Aspergillus oryzae.

Cited by 31 publications

References 2 publications

Overexpressing transcriptional regulator in Chaetomium globosum activates a silent biosynthetic pathway: evaluation of shanorellin biosynthesis

Overexpressing transcriptional regulator in Chaetomium globosum activates a silent biosynthetic pathway: evaluation of shanorellin biosynthesis

Molecular Cloning and Determination of the Nucleotide Sequence of Raw Starch Digesting α-Amylase from Aspergillus awamori KT-11

Cloning of the α-Amylase cDNA ofAspergillus shirousamiiand Its Expression inSaccharomyces cerevisiae

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