Cloning, characterization, and expression of two ?-amylase genes from Aspergillus niger var. awamori

Korman, D. R.; Bayliss, Frank; Barnett, Christopher C.; Carmona, Cynthia L.; Kodama, Katherine H.; Royer, Theresa J.; Thompson, Sheryl A.; Ward, Michael P.; Wilson, Lori J.; Berka, Randy M.

doi:10.1007/bf00312611

Cited by 56 publications

(19 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Overall, AsTaaG1 showed significant sequence identity to other Aspergilli α-amylases. AsTaaG1 showed 97.4% identity to Aspergillus awamori AmyA (Korman et al, 1990), 98.4% to A. flavus AFLA_026140A (Payne et al, 2006), 78.2% to Aspergillus fumigatus amylase (Nierman et al, 2005), 97.8% to Aspergillus kawachii AmyA (Nagamine et al, 2003), 69.5% to Aspergillus niger An04g06930 (Pel et al, 2007), 69.5% to A. nidulans AN2018.2 (Galagan et al, 2005), 98.2% to A. oryzae TaaG1 (Tsukagoshi et al, 1989), 97.8% to A. oryzae TaaG2 (Tsukagoshi et al, 1989), 97.6% to Aspergillus shirousami Amy (Shibuya. et al, 1992).…”

Section: Amplification Of the Astaag1 Gene And Its Cdna By Pcr And Rtmentioning

confidence: 99%

Molecular Analysis of the α-Amylase Gene, AstaaG1, from Shoyu Koji Mold, Aspergillus sojae KBN1340

Yoshino-Yasuda¹,

Fujino²,

Matsui³

et al. 2013

FSTR

View full text Add to dashboard Cite

Aspergillus sojae generally has only one ortholog of the Aspergillus oryzae taa (α-amylase) gene. The AstaaG1 gene from a shoyu koji mold, A. sojae KBN1340, comprised 2,063 bp with eight introns. AsTaaG1 consisted of 498 amino acid residues possessing high identity to other Aspergilli α-amylase sequences. Disruption of the AstaaG1 gene resulted in no detectable α-amylase production in starch medium. Promoter activity of the AstaaG1 gene, monitored by xylanase activity, was upregulated with replacement of the CCAAT-like sequence. Site-directed mutation of the CCAAT-like sequence increased xylanase production approximately four times higher than that of the wild type. These results clearly demonstrate that the decreased copy number of the taa gene and the low affinity binding sequence to the Hap complex lead to the lower amylolytic activity of A. sojae compared to that of A. oryzae.

show abstract

Section: Amplification Of the Astaag1 Gene And Its Cdna By Pcr And Rtmentioning

confidence: 99%

Molecular Analysis of the α-Amylase Gene, AstaaG1, from Shoyu Koji Mold, Aspergillus sojae KBN1340

Yoshino-Yasuda¹,

Fujino²,

Matsui³

et al. 2013

FSTR

View full text Add to dashboard Cite

show abstract

“…Since the N-terminal amino acid was not methionine but leucine, it turns out that N-terminal of Amyl III was processed. When the identified N-terminal amino acid sequence was compared to the existing sequences in the database, acid α-amylase of A. niger (Korman et al, 1990) showed 7 identical residues. Internal amino acid sequences of the fragments produced by peptidase digestion of Amyl III were TITYDWDADLV and SLSDALHRGMWL.…”

Section: Resultsmentioning

confidence: 99%

“…Amino acid sequence similarity The amino acid sequence of Amyl III has 93% homology with the acid α-amylase from A. niger (Korman et al, 1990). On the other hand, the homology with α-amylase from Aspergillus sp.…”

Section: Resultsmentioning

confidence: 99%

“…The nucleotide sequences and the amino acid sequences of α-amylases from Aspergillus sp. have been reported (Tada et al, 1989;Wirsel et al, 1989;Korman et al, 1990), however almost all of these α-amylases cannot hydrolyze raw starch. The acid stable α-amylase from A. kawachii is the sole raw-starch-digesting α-amylase so far reported from Aspergillus sp.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

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

View full text Add to dashboard Cite

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.

show abstract

“…niger var. awarnori [44] which show very high sequence identity in both exons and introns. The three highly similar Mucor racernosus EF-la genes share one intron that also shows a high sequence identity, whereas a second intron, that is present at identical positions but is not conserved in sequence, is found in only two of these genes [45].…”

Section: Discussionmentioning

confidence: 99%

The polygalacturonases of Aspergillus niger are encoded by a family of diverged genes

Bussink

Buxton

Fraaye

et al. 1992

European Journal of Biochemistry

117

View full text Add to dashboard Cite

Aspergillus niger produces several polygalacturonases that, with other enzymes, are involved in the degradation of pectin. One of the two previously characterized genes coding for the abundant polygalacturonases T and I1 (PGI and PGII) found in a commercial pectinase preparation was used as a probe to isolate five more genes by screening a genomic DNA library in phage lEMBL4 using conditions of moderate stringency. The products of these genes were detected in the culture medium of Aspergilh nidulans transformants on the basis of activity measurements and Western-blot analysis using a polyclonal antibody raised against PGI. These transformants were, with one exception, constructed using phage DNA. A . niduluns transformants secreted high amounts of PGI and PGII in comparison to the previously characterized A. niger transformants and a novel polygalacturonase (PGC) was produced at high levels by A. nidulans transformed with the subcloned pguC gene. This gene was sequenced and the protein-coding region was found to be interrupted by three introns; the different intronlexon organization of the three sequenced A. niger polygalacturonase genes can be explained by the gain or loss of two single introns. The pgaC gene encodes a putative 383-amino-acid prepro-protein that is cleaved after a pair of basic amino acids and shows approximately 60% amino acid sequence similarity to the other polygalacturonases in the mature protein. The N-terminal amino acid sequences of the A. niger polygalacturonases display characteristic amino acid insertions or deletions that are also observed in polygalacturonases of phytopathogenic fungi. In the upstream regions of the A . niger polygalacturonase genes, a sequence of ten conserved nucleotides comprising a CCAAT sequence was found, which is likely to rcpresent a binding site for a regulatory protein as it shows a high similarity to the yeast CYCl upstream activation site recognized by the HAP2/3/4 activation complex.Polygalacturonases are involved in the degradation orpectin, a complex polysaccharide that is primarily found in the middle lamella and primary cell wall of higher plants [I]. Polygalacturonases as well as other pectolytic enzymes are produced by many organisms (reviewed in [2]), and microbial pectolytic enzymes have been extensively studied in relation to plant pathogenesis. The molecular biology of the bacterial enzymes, especially the pectate lyases of Erwiniu, is well developed compared to that of the pectolytic enzymes of fungal pathogens [3, 41. Recently, genomic or cDNA clones coding for pectinesterase [5], pectin lyases [6, 71, pectate lyase [8] and polygalacturonases [9 -121 have been obtained from Aspei-gillus species, which are considered to be saprophytes. The availability of commercial pectinase preparations derived from Aspergillus niger has contributed to the rapid progress in gene Correspondence to J. Visser, Section

show abstract

Cloning, characterization, and expression of two ?-amylase genes from Aspergillus niger var. awamori

Cited by 56 publications

References 24 publications

Molecular Analysis of the α-Amylase Gene, AstaaG1, from Shoyu Koji Mold, Aspergillus sojae KBN1340

Molecular Analysis of the α-Amylase Gene, AstaaG1, from Shoyu Koji Mold, Aspergillus sojae KBN1340

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

The polygalacturonases of Aspergillus niger are encoded by a family of diverged genes

Contact Info

Product

Resources

About