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A method for improving the thermal stability of linear maltooligosaccharide producing enzyme

A technology of maltooligosaccharides and thermal stability, which is applied in the field of enzyme engineering, can solve problems affecting the normal progress of the reaction, achieve the effects of improving thermal stability, low price, and increasing industrial application value

Active Publication Date: 2020-06-09
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently commonly used stabilizers include sugars and polyols, such as trehalose, sucrose, polyethylene glycol, etc., but the addition of the above stabilizers sometimes affects the normal progress of the reaction

Method used

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  • A method for improving the thermal stability of linear maltooligosaccharide producing enzyme
  • A method for improving the thermal stability of linear maltooligosaccharide producing enzyme
  • A method for improving the thermal stability of linear maltooligosaccharide producing enzyme

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Straight-chain maltopentaose-hexaose-generating enzyme from Bacillus stearothermophilus STB04 (Genbank accession number: AIV43245.1) was combined with CaCl 2 The solution is mixed homogeneously so that Ca 2+ The final concentrations were 1, 2, 5, 10, 20mM respectively, and the enzyme concentration was 20U / mL. The thermal stability of the enzyme solution was measured at 80°C, and the results were as follows: figure 1 As shown, the half-life of the straight-chain maltopentaose-hexaose generating enzyme at 80°C was less than 5min. Compared with the blank control, adding Ca 2+ Afterwards, its thermal stability was significantly improved, and the final concentration of 1mM Ca 2+ The half-life of the enzyme at 80°C can be extended to more than 30 minutes, and the final concentration of 10mM Ca 2+ The best effect can increase the half-life to more than 80min.

Embodiment 2

[0026] Add NaCl solution to the linear maltopentaose-hexasaccharide generating enzyme solution derived from Bacillus stearothermophilus STB04 to make Na + The final concentration is 5-80mM, and the final concentration of the enzyme is 100U / mL. The thermal stability of the enzyme solution is measured at 80°C, and the results are as follows: figure 2 As shown, compared with the blank control, the addition of Na + Afterwards, its thermal stability was significantly improved, and the final concentration of 5mM Na + The half-life of the enzyme can be extended to 20min at 80°C with a final concentration of 40mM Na + The half-life of the enzyme can be extended to 69min at 80°C.

Embodiment 3

[0028] Add CaCl 2 solution makes Ca 2+ The final concentration was 10mM, and NaCl solution was added to make Na + The final concentration is 40mM, and the final concentration of enzyme is 20U / mL. The thermal stability of the enzyme solution was measured at 80°C, and the results were as follows: image 3 As shown, compared with the blank control without adding additives, only adding 10mM Ca 2+ or 40mM Na + Compared with the control, the thermal stability of linear maltohexaose generating enzyme was significantly improved. After being incubated at 80°C for 3 hours, its relative activity still reached 71.1%, which was 4.7 times that of adding calcium ions alone, and 5.8 times that of adding sodium ions alone. The blank control without additives was completely inactivated.

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Abstract

The invention discloses a method for improving the heat stability of linear maltooligosacchaeide generating enzyme, belonging to the technical field of enzyme engineering. By synergetically adding calcium ions and sodium ions, the heat stability of the linear maltooligosacchaeide generating enzyme is remarkably improved, the relatively activity of the enzyme after being subjected to temperature maintenance at 60-90 DEG C for 30min-3h is 2-10 times of that of the enzyme which is not added with an aid and is increased by at least two times relatively to the enzyme which is independently added with Ca<2+> or Na<+>. The method has very important significance to the increase of the industrial application value of the enzyme. Furthermore, the method is simple in operation, an adopted reagent islow in cost, safe and non-toxic, and the method has very good application prospects.

Description

technical field [0001] The invention relates to a method for improving the thermal stability of an enzyme producing linear maltooligosaccharides, and belongs to the technical field of enzyme engineering. Background technique [0002] Linear maltooligosaccharide-forming enzymes refer to a class of enzymes that can hydrolyze starch to produce different types of linear maltooligosaccharides, and are widely used in food, medicine, textile, paper and other industries. During the enzymatic production of linear maltooligosaccharides, the thermostability of the enzyme is a key factor. The high thermal stability of the enzyme is not only beneficial to prolong the storage time of the enzyme, reduce the loss of enzyme activity during storage and transportation, but also enable the enzyme to maintain a higher activity at a higher temperature, thereby improving the reaction efficiency and shortening the production time. cycle, thereby reducing production costs. [0003] The methods tha...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/96
CPCC12N9/00C12N9/96
Inventor 李兆丰顾正彪潘思惠李才明程力洪雁
Owner JIANGNAN UNIV