Cellulase gene
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Solution Overview
Problem
The challenge is to identify and isolate endoglucanase and β-glucosidase genes from Acremonium cellulolyticus to optimize the composition of cellulase components produced by this microorganism, as only two types of cellobiohydrolases and one type of β-glucosidase have been isolated, limiting the potential for enhanced expression or disruption using gene recombination techniques.
Innovation Solution
The approach involves comparing amino acid sequences of known endoglucanases and β-glucosidases to find conserved regions, designing primers, and using PCR to amplify and sequence gene fragments, ultimately leading to the identification of specific endoglucanase and β-glucosidase genes, which can be used to produce recombinant proteins and microorganisms with enhanced cellulase activity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If gene recombination techniques are used to overexpress specific enzyme components, then cellulase activity can be optimized for specific uses, but the technique cannot be applied because most endoglucanase and β-glucosidase genes have not been isolated
Solution Approach 1:
The patent applies preliminary action by isolating and characterizing multiple endoglucanase and β-glucosidase genes from Acremonium cellulolyticus before gene recombination can be performed. The systematic isolation of genes ACC3, ACC5, ACC6, ACC7, ACC8, ACC9, ACC10, BGLC, and BGLD creates the necessary genetic resources that enable subsequent overexpression studies and optimization of cellulase activity for specific industrial applications.
2Adaptability or versatility
If only two types of cellobiohydrolases and one type of β-glucosidase are isolated, then the cellulase composition cannot be optimized using gene recombination, but this limits the ability to enhance cellulase activity for specific uses
Solution Approach 1:
The patent applies segmentation by dividing the cellulase system into distinct gene components (ACC3, ACC5, ACC6, ACC7, ACC8, ACC9, ACC10 for endoglucanases and BGLC, BGLD for β-glucosidases). This gene-level segmentation allows independent manipulation and overexpression of specific enzyme components, enabling tailored optimization of cellulase composition for different industrial applications such as saccharification, textile treatment, or feed additives.
Data Source
AI summary
An object is to identify endoglucanase and β-glucosidase genes by isolating genomic DNA containing cellulase genes, which are classified into endoglucanases or β-glucosidases, from Acremonium cellulolyticus, and sequencing the nucleotide sequences thereof. The inventors intensively compared the amino acid sequences of known endoglucanases and β-glucosidases with each other to find conserved region of amino acid sequences in Acremonium cellulolyticus, and various primers were designed based on the information. PCR was carried out using the various primers thus designed and genomic DNA or cDNA as a template. As a result, gene fragments of endoglucanases and β-glucosidases were obtained. Primers were designed based on the gene fragments, and PCR was carried out to amplify nine genes of endoglucanases and β-glucosidases. The nucleotide sequences thereof were sequenced, and the present invention was completed.