Sequence Analysis and Overexpression of a Pectin Lyase Gene (pel1) from Aspergillus oryzae KBN616

Kitamoto, Noriyuki; Yoshino-Yasuda, Shoko; Ohmiya, Kunio; Tsukagoshi, Norihiro

doi:10.1271/bbb.65.209

Cited by 40 publications

(27 citation statements)

References 12 publications

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“…6C). Expression of the antisense pepB led to a reduction in the presence of aspergillopepsin B ranging from 10% in transformant asX4 to 70% in transformant asH5, values similar to those obtained in Aspergillus with other antisense strategies (6,24,31,40). However, the small increment in the increase of thaumatin production observed after antisense RNA formation indicated that the remaining aspergillopepsin B still degraded thaumatin; therefore, it was necessary to disrupt the pepB gene in the thaumatin-producing strain TB2b1-44-pyrG45 to remove completely aspergillopepsin B from culture broths.…”

Section: Discussionsupporting

confidence: 78%

Silencing of the Aspergillopepsin B ( pepB ) Gene of Aspergillus awamori by Antisense RNA Expression or Protease Removal by Gene Disruption Results in a Large Increase in Thaumatin Production

Moralejo

Cardoza

Gutiérrez

et al. 2002

Appl Environ Microbiol

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Section: Discussionsupporting

confidence: 78%

Silencing of the Aspergillopepsin B ( pepB ) Gene of Aspergillus awamori by Antisense RNA Expression or Protease Removal by Gene Disruption Results in a Large Increase in Thaumatin Production

Moralejo

Cardoza

Gutiérrez

et al. 2002

Appl Environ Microbiol

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“…The native xlnD contains a predicted signal peptide of 26 or 27 amino acids, and the mature protein has a predicted molecular mass of 88 kDa. It also exhibits significant levels of similarity at the amino acid level to the A. nidulans (66% identity), A. oryzae (64% identity), and T. reesei (63% identity) ␤-xylosidases (20,33,35). It is noteworthy that the Bxl1 of T. reesei also exhibited ␣-L-arabinofuranosidase and ␣-L-arabinopyranosidase activities (33).…”

Section: Discussionmentioning

confidence: 98%

“…The protein is N glycosylated and contains 15 potential Nglycosylation sites (48). Sequence similarity was found to ␤-glycosidases (␤-xylosidase and ␤-glucosidases) of family 3, which include enzymes from both bacterial and fungal origins (20,33,35,48). The condensation reaction of this ␤-xylosidase has been used for the synthesis of disaccharides such as ␤,␤-1,1-xylodisaccharide, ␤-1,4-xylodisaccharide (xylobiose), ␤-1,2-xylodisaccharide, ␣-1,4-xylodisaccharide, and ␤-1,3-xylodisaccharide (17).…”

mentioning

confidence: 99%

Degradation of Xylan to d -Xylose by Recombinant Saccharomyces cerevisiae Coexpressing the Aspergillus niger β-Xylosidase ( xlnD ) and the Trichoderma reesei Xylanase II ( xyn2 ) Genes

Grange

Pretorius

Claeyssens

et al. 2001

Appl Environ Microbiol

117

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The ␤-xylosidase-encoding xlnD gene of Aspergillus niger 90196 was amplified by the PCR technique from first-strand cDNA synthesized on mRNA isolated from the fungus. The nucleotide sequence of the cDNA fragment was verified to contain a 2,412-bp open reading frame that encodes a 804-amino-acid propeptide. The 778-amino-acid mature protein, with a putative molecular mass of 85.1 kDa, was fused in frame with the Saccharomyces cerevisiae mating factor ␣1 signal peptide (MF␣1 s ) to ensure correct posttranslational processing in yeast. The fusion protein was designated Xlo2. The recombinant ␤-xylosidase showed optimum activity at 60°C and pH 3.2 and optimum stability at 50°C. The K i(app) value for D-xylose and xylobiose for the recombinant ␤-xylosidase was determined to be 8.33 and 6.41 mM, respectively. The XLO2 fusion gene and the XYN2 ␤-xylanase gene from Trichoderma reesei, located on URA3-based multicopy shuttle vectors, were successfully expressed and coexpressed in the yeast Saccharomyces cerevisiae under the control of the alcohol dehydrogenase II gene (ADH2) promoter and terminator. These recombinant S. cerevisiae strains produced 1,577 nkat/ml of ␤-xylanase activity when expressing only the ␤-xylanase and 860 nkat/ml when coexpressing the ␤-xylanase with the ␤-xylosidase. The maximum ␤-xylosidase activity was 5.3 nkat/ml when expressed on its own and 3.5 nkat/ml when coexpressed with the ␤-xylanase. Coproduction of the ␤-xylanase and ␤-xylosidase enabled S. cerevisiae to degrade birchwood xylan to D-xylose.

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“…XYNII represents more than 50% of the total xylanolytic activity of T. reesei cultivated on xylan (39). Meanwhile, the industrial koji mold Aspergillus oryzae is also known to be an efficient producer of xylandegrading enzymes and secretes ␤-xylosidase (21). The gene encoding the major ␤-xylosidase, XylA, comprises 2,397 bp with no intron and encodes a protein consisting of 798 amino acids.…”

mentioning

confidence: 99%

Construction of a Xylan-Fermenting Yeast Strain through Codisplay of Xylanolytic Enzymes on the Surface of Xylose-Utilizing Saccharomyces cerevisiae Cells

Katahira¹,

Fujita²,

Mizuike

et al. 2004

Appl Environ Microbiol

151

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Hemicellulose is one of the major forms of biomass in lignocellulose, and its essential component is xylan. We used a cell surface engineering system based on ␣-agglutinin to construct a Saccharomyces cerevisiae yeast strain codisplaying two types of xylan-degrading enzymes, namely, xylanase II (XYNII) from Trichoderma reesei QM9414 and ␤-xylosidase (XylA) from Aspergillus oryzae NiaD300, on the cell surface. In a high-performance liquid chromatography analysis, xylose was detected as the main product of the yeast strain codisplaying XYNII and XylA, while xylobiose and xylotriose were detected as the main products of a yeast strain displaying XYNII on the cell surface. These results indicate that xylan is sequentially hydrolyzed to xylose by the codisplayed XYNII and XylA. In a further step toward achieving the simultaneous saccharification and fermentation of xylan, a xylan-utilizing S. cerevisiae strain was constructed by codisplaying XYNII and XylA and introducing genes for xylose utilization, namely, those encoding xylose reductase and xylitol dehydrogenase from Pichia stipitis and xylulokinase from S. cerevisiae. After 62 h of fermentation, 7

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Sequence Analysis and Overexpression of a Pectin Lyase Gene (pel1) from Aspergillus oryzae KBN616

Cited by 40 publications

References 12 publications

Silencing of the Aspergillopepsin B ( pepB ) Gene of Aspergillus awamori by Antisense RNA Expression or Protease Removal by Gene Disruption Results in a Large Increase in Thaumatin Production

Silencing of the Aspergillopepsin B ( pepB ) Gene of Aspergillus awamori by Antisense RNA Expression or Protease Removal by Gene Disruption Results in a Large Increase in Thaumatin Production

Degradation of Xylan to d -Xylose by Recombinant Saccharomyces cerevisiae Coexpressing the Aspergillus niger β-Xylosidase ( xlnD ) and the Trichoderma reesei Xylanase II ( xyn2 ) Genes

Construction of a Xylan-Fermenting Yeast Strain through Codisplay of Xylanolytic Enzymes on the Surface of Xylose-Utilizing Saccharomyces cerevisiae Cells

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