Trichoderma fungus having mutant-type bxl1 gene and method for producing xylooligosaccharide and glucose by using same

A Trichoderma, mutant technology, applied in the field of manufacturing xylo-oligosaccharides and glucose, can solve the problem of unknown function of Fn3-like domain, and achieve the effect of high-efficiency manufacturing, high-efficiency glucose and xylo-oligosaccharides

Active Publication Date: 2018-12-21
TORAY IND INC
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, β-xylosidase 1 has an Fn3-like domain in addition to...

Method used

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  • Trichoderma fungus having mutant-type bxl1 gene and method for producing xylooligosaccharide and glucose by using same
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  • Trichoderma fungus having mutant-type bxl1 gene and method for producing xylooligosaccharide and glucose by using same

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Embodiment

[0063] The following examples will be given to specifically describe the present invention. However, the present invention is not limited to these examples.

reference example 1

[0064] Reference Example 1 Protein Concentration Determination Method

[0065] A commercially available protein concentration measurement reagent (Quick Start Bradford Protein Assay, manufactured by Bio-Rad) was used. 5 μL of the diluted filamentous fungus-derived cellulase solution was added to 250 μL of the protein concentration measurement reagent returned to room temperature, and after standing at room temperature for 5 minutes, the absorbance at 595 nm was measured with a microplate reader. Protein concentrations were calculated against a standard curve using BSA as a standard.

reference example 2

[0066] Reference example 2β-xylosidase activity assay method

[0067] β-Xylosidase activity measurement method Specifically, 10 μL of enzyme dilution solution was used in 90 μL of 50 mM acetate buffer containing 1 mM p-nitrophenyl-β-xylopyranoside (manufactured by Sigma Aldrich Japan) at 30°C. After 30 minutes of reaction, add 10 μL of 2M sodium carbonate and mix well to stop the reaction, and measure the increase of absorbance at 405 nm. The activity of liberating 1 μmol of p-nitrophenol per minute was defined as 1U. For the blank, 10 μL of 2M sodium carbonate was added to 90 μL of 50 mM acetate buffer containing 1 mM p-nitrophenyl-β-xylopyranoside and mixed well, then 10 μL of enzyme diluent was added and reacted at 30°C for 30 minutes. Then, the increase in absorbance at 405 nm was measured.

[0068] Reference example 2β-glucosidase activity assay method

[0069] β-glucosidase activity measurement method Specifically, 10 μL of enzyme diluent was used in 90 μL of 50 mM ac...

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Abstract

Disclosed are a novel Trichoderma reesei fungus with which xylan in a biomass is not decomposed to xylose even when a cellulase derived from a Trichoderma reesei fungus is used for the hydrolysis of acellulose-based biomass; and a method for producing glucose and xylooligosaccharide from a cellulose-containing biomass by using said Trichoderma reesei fungus. This Trichoderma fungus includes an N-terminal domain and a C-terminal domain of a beta-xylosidase 1 (BXL1) gene, wherein a Fn3-like domain is disrupted. By using this Trichoderma fungus, beta-xylosidase activity is deleted and beta-glucosidase activity is increase. Thus, xylan in a biomass is not decomposed to xylose even when a cellulose-containing biomass is hydrolyzed, and glucose and xylooligosaccharide can be produced efficiently.

Description

technical field [0001] The present invention relates to a Trichoderma fungus having a mutant BXL1 gene lacking in β-xylosidase (BXL1) activity and a method for producing xylo-oligosaccharides and glucose from cellulosic biomass using the fungus. Background technique [0002] Xylooligosaccharides are a general term for oligosaccharides formed by combining multiple xylose units through β-glycosidic bonds. Xylo-oligosaccharides are also used as materials for functional foods because they exhibit an excellent gut-regulating action (Non-Patent Document 1). [0003] Xylo-oligosaccharides can be obtained by hydrolyzing xylan contained in cellulosic biomass. As hydrolysis methods, hydrothermal treatment (Non-Patent Document 2), acid hydrolysis (Non-Patent Document 3), Enzyme treatment method (Patent Document 1). [0004] On the other hand, when biomass is hydrolyzed by enzymes, filamentous bacteria are very suitable because of their high production capacity of cellulase, an enzyme...

Claims

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Application Information

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IPC IPC(8): C12N15/09C12N1/14C12N1/15C12N15/01C12P19/14C13K1/02C13K13/00
CPCC12P19/14C12N1/14C13K1/02C13K13/00C12N15/01C12N15/09C12N9/2402C12Y302/01037C12N1/145C12R2001/885C12P19/04
Inventor 小林宏治平松绅吾山田胜成
Owner TORAY IND INC
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