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High-temperature-resistant bacillus licheniformis capable of producing alpha-amylase

A technology of Bacillus licheniformis and amylase, applied in the direction of bacteria, microorganism-based methods, microorganisms, etc., can solve the problems of engineering strains that are not resistant to high temperature

Active Publication Date: 2015-02-18
INST OF MICROBIOLOGY HEILONGJIANG ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention aims to solve the problem that the existing α-amylase-producing engineering strains are not resistant to high temperature, and provides a high-temperature-resistant Bacillus licheniformis producing α-amylase

Method used

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  • High-temperature-resistant bacillus licheniformis capable of producing alpha-amylase

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specific Embodiment approach 1

[0018] Specific Embodiment 1: In this embodiment, Bacillus licheniformis HWyb1401 is deposited in the General Microorganism Center of China Committee for the Collection of Microorganisms, and the preservation number is CGMCC No.8718.

[0019] In this embodiment, Bacillus licheniformis (Bacillus licheniformis) HWyb1401 is isolated from compost manure in Gannan County, Qiqihar City.

[0020] Filtering is done in the following steps:

[0021] Weigh 10.0g of compost fertilizer and put it into a 100ml sterile water triangular flask containing glass beads, and shake it at 200r / min for 30min to make a bacterial suspension; take 1ml of the suspension and add it to 100mL beef extract peptone medium, at 40 Under the conditions of ℃ and 180r / min, shake culture for 18h-20h, then carry out gradient dilution, spread on the starch agar powder screening medium, put it in the environment of 40℃ and culture for 48h, select the single colony plate of each strain, and use Lu Cover the plate with...

specific Embodiment approach 2

[0023] Specific embodiment two: In this embodiment, Bacillus licheniformis (Bacillus licheniformis) HWyb1401 is inoculated in beef extract peptone fermentation medium with an inoculum size of 1%, cultivated at a temperature of 55°C and 200r / min for 72h, and then at 10000rr / min, refrigerated and centrifuged at 4°C for 30 min, and the supernatant was collected to measure amylase activity by iodine colorimetry.

[0024] The amylase activity in the fermentation broth of Bacillus licheniformis HWyb1401 was 70U / mL.

[0025] In the present embodiment, the starch hydrolysis circle of Bacillus licheniformis (Bacillus licheniformis) HWyb1401 is as follows: figure 1 shown.

specific Embodiment approach 3

[0026] Specific embodiment three: In this embodiment, Bacillus licheniformis (Bacillus licheniformis) HWyb1401 is inoculated in the beef extract peptone fermentation medium under different temperature conditions according to the inoculum size of 1%, and cultivated under the condition of 200r / min for 72h; then at 10000r / min refrigerated and centrifuged at 4°C for 30 min, and the supernatant was collected to measure amylase activity by iodine colorimetry.

[0027] Experiments have found that they are active and produce α-amylase under the culture conditions of 55°C to 70°C.

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Abstract

The invention relates to high-temperature-resistant bacillus licheniformis capable of producing alpha-amylase and relates to bacillus licheniformis, solving the problem that existing engineering strains for producing alpha-amylase are unable to resist high temperature. The high-temperature-resistant bacillus licheniformis is bacillus licheniformis HWyb1401 which is preserved in CGMCC with the preservation number of CGMCC No. 8718. The bacillus licheniformis HWyb1401 provided by the invention has a high-temperature-resistant characteristic and still can survive at 70 DEG C and can be metabolized to generate alpha-amylase. The enzyme activity of the alpha-amylase in a fermentation liquid at 55 DEG C can reach 70U / mL.

Description

technical field [0001] The invention relates to a strain of Bacillus licheniformis. Background technique [0002] α-amylase can hydrolyze α-1,4 glycosidic bonds from the non-reducing end of the glucose chain, and can endocut randomly, and the product is a mixture of glucose, maltose, maltotriose and α-cyclodextrin. α-amylase has excellent high temperature resistance performance, especially high temperature resistant α-amylase can be used under the condition of 90°C~95°C, and the instantaneous temperature in the jet liquefaction process reaches 105°C~110°C, high temperature resistant α-amylase It can still effectively hydrolyze starch. α-amylase is widely used in starch, alcohol, beer, monosodium glutamate, brewing, textile desizing and other industries due to its excellent high temperature resistance. [0003] Although α-amylase can withstand high temperature, the current engineering strains producing α-amylase are not resistant to high temperature. If the purified α-amyl...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/10
CPCC12N1/205C12R2001/10
Inventor 郭丽姝牛彦波孙宇峰曹亚彬殷博安琦杨旭升樊川迟君道张欣王慧民吴皓琼刘雷明王玉文
Owner INST OF MICROBIOLOGY HEILONGJIANG ACADEMY OF SCI
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