Glucose-resistant beta-glucosidase as well as expression gene and application thereof

A technology of glucosidase and glucose, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problems of waste or incineration, low utilization rate, environmental pollution, etc.

Pending Publication Date: 2022-01-28
HAINAN UNIVERSITY
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

my country is a traditional agricultural country, and straw resources are very rich, but the utilization rate is e

Method used

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  • Glucose-resistant beta-glucosidase as well as expression gene and application thereof
  • Glucose-resistant beta-glucosidase as well as expression gene and application thereof
  • Glucose-resistant beta-glucosidase as well as expression gene and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] (1) Experimental plan

[0057] 1. Cloning and sequence analysis of the bgl gene of Aspergillus polypolyphaga

[0058] 1.1 Polymerum RNA-seq sequencing

[0059] Polypolygonum was inoculated in the fermentation medium and cultured for 5 days to extract samples for RNA-seq. The sample names are marked as in the table, and the samples sent were prepared in three independent repetitions.

[0060] Table 1

[0061]

[0062] 1.2 Cloning and sequence analysis of bgAs1 gene from Aspergillus polypolyphaga

[0063] Analyze the results after RNA-seq, screen the glycoside hydrolase family genes according to the condition of up-regulated gene expression and log2(FC)>2.0, and analyze the sequence of the screened genes, and verify by RT-PCR and Q-PCR its gene expression.

[0064] 1.2.1 Primer design for bgl gene

[0065] The nucleotide sequence of bgAs1 was analyzed, and homologous sequences at both ends of the linearized pPICZαA vector were introduced into the 5' ends of the f...

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Abstract

The invention relates to the technical field of biology, and in particular, relates to glucose-resistant beta-glucosidase as well as an expression gene and application thereof. Aspergillus sydowii L1 capable of producing beta-glucosidase when glucose is used as a substrate is screened from mangrove forest soil, genes bgAs1 with similar expression quantities when glucose, microcrystalline cellulose and cellobiose are used as unique carbon sources for induction are screened by a transcriptome comparison method, in-vitro expression is carried out by a pichia pastoris expression system, and the recombinant beta-glucosidase BgAs1 is obtained after purification. The beta-glucosidase has relatively high glucose tolerance, salt tolerance and wide substrate specificity, shows the potential of replacing a commercial enzyme Nov188 when being cooperated with cellulase to hydrolyze BgAs1 on bagasse, and has a good industrial application prospect.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a glucose-tolerant beta-glucosidase, its expression gene and application. Background technique [0002] Cellulose, as an abundant renewable energy source on earth, has great development potential. Cellulose is a macromolecule formed by connecting glucosides with β-1,4 glycosidic bonds. As the main component of plant polysaccharides, cellulose plays an indispensable role in plant structure and interacts with hemicellulose and lignin. Cross-links constitute the main skeleton of the cell wall. my country is a traditional agricultural country, and straw resources are very rich, but the utilization rate is extremely low, and they are often discarded or burned in agricultural practice, thus causing environmental pollution. The rapid development of animal husbandry has increased the pressure on food, and my country is facing a situation where humans and animals compete for food. This prob...

Claims

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Application Information

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IPC IPC(8): C12N9/42C12N15/56C12N15/70C12N15/81C12N1/21C12N1/19C12P19/02C12P19/04C12P19/12C12P19/14C12P7/10C12R1/19C12R1/84
CPCC12N9/2445C12N15/70C12N15/815C12P19/02C12P19/04C12P19/12C12P19/14C12P7/10C12P2203/00Y02E50/10
Inventor 王学梅刘晓烜孙楠张冰溪张海文管庆丰
Owner HAINAN UNIVERSITY
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