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Mutant gene of xylanase in high acid-base reaction range and point mutation process thereof

A technology of xylanase and mutant gene, applied in genetic engineering, plant genetic improvement, botanical equipment and methods, etc., can solve the problem that the acid-base reaction range of xylanase cannot be effectively improved, and achieve the improvement of acid-base resistance. receptive effect

Inactive Publication Date: 2008-10-22
NAT PINGTUNG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional point mutation method can only improve the acid-resistant properties of xylanase, but still cannot effectively improve the acid-base reaction range of xylanase

Method used

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  • Mutant gene of xylanase in high acid-base reaction range and point mutation process thereof
  • Mutant gene of xylanase in high acid-base reaction range and point mutation process thereof
  • Mutant gene of xylanase in high acid-base reaction range and point mutation process thereof

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Effect test

Embodiment 1

[0027] see figure 1 As shown, Example 1 of the present invention mainly utilizes a one-point mutation method to mutate the 41st amino acid of a xylanase gene from asparagine to aspartic acid to form a polyxylanase with a high acid-base reaction range. Mutant gene for carbohydrase. More specifically, the present invention uses the point mutation method to further form a mutant gene of xylanase with a high acid-base reaction range through the following steps:

[0028] The first step S1 is to produce the xylanase gene using a vector to form a first recombinant plastid, and then transform the first recombinant plastid into a competent cell to further form a wild type expression vector;

[0029] The second step S2 is to mix the first recombinant plasmid with a dNTP, a buffer solution [reactionbuffer], a forward primer [forward primer], a reverse primer [reverse primer] and a polymerase [ polymerase] carry out a polymerase chain reaction [polymerase chain reaction] to form a seco...

Embodiment 2

[0044] see figure 2 and Figure 7 Shown, the gene sequence of the mutant gene of xylanase of embodiment 2 of the present invention is to contain such as Figure 7 The sequence of [SEQ ID NO: 4, SEQ ID NO: 4] is shown.

[0045] Figure 7 In , the region marked with a box line is the mutation position, which is the 21st amino acid of Sequence ID number: 4, and is aspartic acid. Wherein, because the sequence identification number: 4 is the gene sequence formed for the 21st to 200th amino acids in the sequence identification number: 3. Furthermore, it can be understood that although the mutated positions are different from the positions of Sequence Identification Number: 3 and Sequence Identification Number: 4, the positions of the mutated amino acids are substantially the same. Therefore, it can be found that the mutant gene of xylanase containing the gene sequence of SEQ ID NO: 4 also has the effect of improving the acid-base tolerance, acid-base reaction range and activity...

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Abstract

The invention relates to a xylanase mutant gene with a high acid-base reaction range and a point mutation method for the same. The invention is to utilize the point mutation method to mutate the forty-first amino acid of a xylanase gene from asparagines to asparagus amino acids so as to form the xylanase mutant gene with the high acid-base reaction range. Therefore, the method can effectively improve the acid-base tolerance of xylanase, the acid-base reaction range and the activity.

Description

technical field [0001] The present invention relates to a mutant gene of xylanase in a high acid-base reaction range and a point mutation method thereof, which uses a point mutation method to convert the 41st amino acid of a xylanase gene from asparagine [asparagine 〕mutated to aspartic acid [aspartic acid] to form a mutant gene of xylanase with a high acid-base reaction range. Background technique [0002] Generally speaking, xylan (xylan) mostly exists in the structural polysaccharides of plants, which has the function of protecting the plant's fiber, so xylan is regarded as an obstacle for human beings to exploit plant resources. For example, in the pulp formed during the papermaking process, xylan will adhere to the lignin of the plant on the surface of the fiber of the plant, and it is usually necessary to bleach the pulp with monochloride. After the bleaching operation is completed, the chemical by-products produced by chlorides are toxic and carcinogenic, and they wi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/56C12N9/24
Inventor 郑雪玲陈又嘉江育全
Owner NAT PINGTUNG UNIV OF SCI & TECH
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