A kind of pseudomonas roseri pr415 and its application

A technology of Pseudomonas roseri and Pseudomonas strains, applied in the field of microorganisms, can solve the problems of limited application range, variability and inactivation, and achieve the effects of high salt tolerance, short time and great application value

Active Publication Date: 2021-11-30
深圳中绿环境集团有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, amylases that can meet the needs of industrial applications are mainly derived from bacteria of the genus Bacillus and fungi of the genus Aspergillus. It has enzymolytic activity only under the conditions of neutrality, acidity and narrow temperature range, and it is very easy to denature and inactivate under alkaline conditions or conditions with large temperature range changes, which seriously limits its application range

Method used

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  • A kind of pseudomonas roseri pr415 and its application
  • A kind of pseudomonas roseri pr415 and its application
  • A kind of pseudomonas roseri pr415 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] 1. Isolation and purification of Pseudomonas roseri PR415

[0029] Weigh 10g of soil sample (taken from the mangrove soil of Hailing Island Mangrove National Wetland Park, Guangdong Province) in the ultra-clean workbench, and add 90ml sterile water to the soil sample, place it on a shaker for 60min, and make the soil The sample is evenly dispersed in the diluent to become a soil suspension; after the soil is dispersed, draw 100ul of the soil suspension into 900ul of sterile water to obtain a 10-fold dilution, and then dilute 10 times in sequence to obtain 100 times, 1000 times, 10000 times, 100,000-fold and 1,000,000-fold dilutions, the entire process is carried out in a clean bench.

[0030] Take 100ul of dilutions with different dilution factors and spread them on three LB solid culture plates, and then place the culture plates in an incubator at 37°C for 2-3 days at a constant temperature until plaques grow on the culture plates.

[0031] According to the growth of ...

Embodiment 2

[0049] Cultivation of Pseudomonas roserii PR415

[0050] The Pseudomonas roseri PR415 seed liquid that above-mentioned embodiment 1 obtains, with 1.0 * 10 6 Individuals / mL inoculum concentration and 1% inoculation volume were inoculated in a 1000ml Erlenmeyer flask containing 300ml LB liquid medium, and cultivated in a constant temperature shaker at 30°C and 180rpm for 48h to obtain the Pseudomonas rosenbergii Bacteria PR415 bacteria solution. After testing, it was found that the obtained bacterial liquid had higher amylase activity.

[0051] The formula and preparation method of LB liquid medium are the same as in Example 1.

Embodiment 3

[0053] Utilize Pseudomonas roseri PR415 isolated in Example 1 to produce amylase

[0054] Inoculation: the seed solution obtained in Example 1 was mixed with 1.0×10 6 Individual / mL inoculum concentration and 1% inoculation volume were inoculated in a 1000ml Erlenmeyer flask containing 300ml LB liquid medium;

[0055] Cultivation: The culture medium inoculated with the above seed liquid was cultured in a constant temperature shaker at 30° C. and 180 rpm for 24 hours to obtain a Pseudomonas roserii PR415 bacterial liquid.

[0056] Centrifugal separation: After the culture is over, centrifuge to remove the bacterial precipitate in the bacterial liquid, and keep the supernatant.

[0057] The supernatant obtained above is the crude enzyme solution containing amylase.

[0058] Wherein, the formula and preparation method of the LB liquid medium are the same as those in Example 1.

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Abstract

The invention relates to the technical field of microorganisms, and specifically discloses a Pseudomonas roserii PR415 and an application thereof. The strain preservation number of the Pseudomonas roserii PR415 is: CCTCC NO: M2021331. The Pseudomonas roserii PR415 can be used to prepare amylase. The Pseudomonas rosenbergii PR415 provided by the present invention breaks through the limitation that the amylase that meets the needs of industrial applications is mainly derived from the genus Bacillus and the genus Aspergillus, and can be fermented to obtain a highly active amylase solution that meets the needs of industrial applications. The obtained amylase solution has a wide pH and temperature adaptation range, and can maintain high enzyme activity under the conditions of pH 4-11 and temperature 30-80°C. Moreover, the Pseudomonas roserii PR415 also has the excellent characteristics of fast growth and reproduction, high salt tolerance and short amylase production time, and has great application value in the fields of industry, agriculture and medicine.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a Pseudomonas roserii PR415 and an application thereof. Background technique [0002] Amylase is the general term for enzymes that hydrolyze starch and glycogen, and are widely used in industries such as food, textile, paper, detergent and agricultural waste treatment. With the discovery and use of more new amylases, the application range of amylases has expanded from traditional industrial fields to clinical, pharmaceutical and agricultural fields. Amylase has a wide range of sources and can be isolated from animals, plants and microorganisms. Among them, amylase derived from microorganisms has the characteristics of abundant sources, diverse performances and easy industrial production, which can meet the needs of various industrial applications and is the most widely used in industry. In the process of starch treatment in modern industry, the hydrolysis method of micro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/26C12R1/38
CPCC12N1/20C12N9/2411
Inventor 王磊黄捷黄雅婷王辉宋雪
Owner 深圳中绿环境集团有限公司
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