Multifunctional fusion enzyme XABT gene and construction method and application thereof

Abstract

The invention discloses a multifunctional fusion enzyme XABT gene and a construction method and application thereof; the multifunctional fusion enzyme XABT gene can simultaneously express xylanase, phytase, glucanase and cellulase and has high enzyme activity; the multifunctional fusion enzyme enhances glucanase activity and, therefore, allows for a greater ability to digest barley, oat, rye, wheat and their bran rations with higher glucan content. The multifunctional fusion enzyme XABT gene can also be used for producing grain-saving environment-friendly transgenic economic animals; main anti-nutritional factors in feed are eliminated, so that the purposes of increasing feed digestibility and reducing pollution emission are achieved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a multifunctional fusion enzyme XABT gene and its construction method and application. Background technique [0002] Feed contains a large amount of cellulose, hemicellulose (arabinoxylan, β-glucan, mannan, etc.), pectin and other non-starch polysaccharide (NSP) anti-nutritional factors, which affect the nitrogen and energy utilization of feed. major antinutritional factor. Phytic acid is the main form of phosphorus in plants. Pigs lack phytase and cannot digest and utilize phosphorus in it, resulting in waste of plant-sourced phosphorus and environmental pollution. [0003] Although the application of non-starch multienzymes and phytases in the feed industry has achieved initial success, there are still a series of problems in their practical production and application. The enzyme preparation industry mainly relies on microbial fermentation to produce industrial enzymes. There are...

AI Technical Summary

Problems solved by technology

In the patent publication number CN106086068A, it is also disclosed that the fusion expression of pectinase-xylanase-phytase-cellulase is realized by using 2A connecting peptide, but the expression efficiency is not high

However, these existing technologies have the following problems: single gene transfer, high expression level, but few types of expressed enzymes, elimination of individual feed nutritional factors has limited improvement in animal performance, and the economic value is not large

2A-mediated multi-gene transfer still has the problems of low co-expression efficiency, inconsistent expression of the first and last genes of the multi-gene fusion enzyme, and difficulty in gene arrangement

However, it is more difficult to choose a suitable connecting peptide, and it is easy to be ignored in the design process of fusion protein

If the functional domains are directly fused without a linker peptide, it will lead to some undesirable results, such as misfolding of the fusion protein, low yield or impaired activity, etc.

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