Beta-mannase mutant Man5AS11R with improved heat resistance and specific activity and encoding gene thereof

A mannanase and specific activity technology, applied in the fields of biotechnology and enzyme engineering, can solve the problems of low enzyme activity, unsatisfactory heat resistance and enzyme activity, and high price
CN110669748AInactive Publication Date: 2020-01-10TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
Publication Date
2020-01-10
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention, which relates to the fields of biotechnology and enzyme engineering, particularly discloses a beta-mannase mutant Man5AS11R with improved heat resistance and specific activity and an encoding gene thereof. The preparation method comprises the following steps: carrying out double mutation of K16C and D296C on beta-mannase Man5A of which the amino acid sequence is shown as SEQ ID NO.1to obtain a mutant Man5AS11R that enables the heat resistance of the mutant to be improved by 5 DEG C by being compared with Man5A; for the Man5ASamino acid sequence shown by SEQ ID NO. 2, carrying out simultaneous mutation of three points of R21V, I80D and I218E continuously to obtain a mutant Man5AS11R with the amino acid sequence shown by SEQ ID NO.4. The obtained mutant Man5AS11R has the activity that is improved by 22.5% and 31.4% respectively be being compared with the Man5A and the Man5AS on the premise that the heat resistance of the mutant Man5AS11R is kept to be unchanged. The over80% of enzyme activity of the Man5AS can be left when the Man5AS is kept for 3min at a high temperature of 80 DEG C but the Man5A can only keep 80% of enzyme activity when being kept for 3min at a temperature of 75 DEG C. Therefore, the beta-mannase mutant Man5AS11R and the encoding gene thereof have the broad development and application prospects.
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Description

technical field

[0001] The invention belongs to the fields of biotechnology and enzyme engineering, and relates to a β-mannanase mutant with improved heat resistance and specific activity and its coding gene. Background technique

[0002] At present, β-mannanase is one of the 12 kinds of feed-grade enzyme preparations approved for use in my country, and is widely used in the feed industry as a high-efficiency feed additive. Since the 1990s, enzyme preparations have been added to 90% of poultry feed in Europe and the United States, and enzyme preparations have been added to 60% of pig rations. my country started late, but it has always been the focus of feed production and application research. As an important feed enzyme, mannanase has received more and more attention in recent years. Adding mannanase to chicken (poultry) feed can effectively increase the growth of broiler chickens, increase the laying rate of laying hens, improve the utilization rate of feed, reduce the ex...

Claims

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