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Strain of bacillus pumilus BP-09 tolerant to high-concentration tea residues and application of bacillus pumilus BP-09

A technology of Bacillus pumilus, BP-09, applied in the direction of application, bacteria, microbial-based methods, etc.

Active Publication Date: 2020-10-27
清远一生自然生物研究院有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Aiming at the deficiencies in the prior art, the technical problem to be solved in the present invention is to use ARTP to mutagenize Bacillus pumilus, to screen high-performance mutants of Bacillus pumilus and to use them in the fermentation of tea dregs and the high-value utilization of tea dregs resources

Method used

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  • Strain of bacillus pumilus BP-09 tolerant to high-concentration tea residues and application of bacillus pumilus BP-09
  • Strain of bacillus pumilus BP-09 tolerant to high-concentration tea residues and application of bacillus pumilus BP-09
  • Strain of bacillus pumilus BP-09 tolerant to high-concentration tea residues and application of bacillus pumilus BP-09

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Experimental program
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Embodiment 1

[0055] The ARTP mutagenesis of embodiment 1 bacterial strain

[0056] 1. Strain activation

[0057] Take the glycerol bacteria of Bacillus pumilus and inoculate them on the slant medium of the bacterial culture medium, and incubate at 30°C for 24 hours. After the culture is over, take a ring of the strain and put it into a fresh slant medium, and incubate at 30°C for 16 hours to further strengthen the vitality of the strain , rejuvenate the strain to achieve the purpose of strain activation.

[0058] 2. Determination of strain ARTP mutagenesis parameters

[0059] Add sterile physiological saline to the slant that has been activated and cultivated, elute, prepare a bacterial suspension, and control the OD of the bacterial suspension 600nm The value is between 0.5-0.7. Take 10 μL of the bacterial suspension and evenly spread it on the surface of the metal slide. After drying, use sterilized forceps to transfer the plate with the sample slide to the ARTP operating chamber. Us...

Embodiment 2

[0064] The domestication screening of embodiment 2 mutant strains

[0065] Take the ARTP-treated bacterial suspension and inoculate it directly into the acclimatization medium, the liquid volume is 50mL of 250mL triangular flask, 30°C, 220rpm, culture for 72h, and investigate the growth of the strain in the medium with tea residue as the main carbon source and reproductive capacity. And by gradually increasing the content of tea residue in the acclimation medium, combined with plate separation, an excellent bacterial strain with strong growth ability, tolerance and utilization of high-concentration tea residue is obtained, preserved, and used for future use. details as follows:

[0066] (1) Domestication of mutant strains by low concentration of tea dregs

[0067] The ARTP-treated bacterial suspension was directly transferred to the acclimation medium for cultivation, and the untreated strain was used as a control to observe the color and odor changes of the fermentation bro...

Embodiment 3

[0077] The identification of embodiment 3 bacillus pumilus BP-09

[0078] 1. Morphological characteristics

[0079] The Bacillus pumilus BP-09 strain was prepared into a bacterial suspension and spread on TSA medium after dilution. After culturing at 30°C for 48 hours, the colonies and cell morphology were observed.

[0080] The results showed that: after being cultured on TSA medium for 48 hours, the colony was round, flat, opaque, light yellow, wrinkled on the surface, and the edges were irregular. Observed under the microscope, the cells were rod-shaped, with blunt rounded ends.

[0081] 2. Physiological and biochemical characteristics

[0082] According to the bacterial identification manual, the physiological and biochemical characteristics of Bacillus pumilus BP-09, such as VP reaction, carbon source utilization, and indole test, were determined.

[0083] The results showed that: the bacterial strain was Gram-positive bacteria, which could not reduce nitrate, the citra...

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Abstract

The invention relates to the technical field of microbial mutation breeding, in particular to a strain of bacillus pumilus BP-09 tolerant to high-concentration tea residues and an application of the bacillus pumilus BP-09. The bacillus pumilus BP-09 is preserved in the Guangdong Microbiological Culture Collection Center (GDMCC) on June 16, 2020, and a preservation number of the bacillus pumilus BP-09 is GDMCC No: 61062. In order to prepare functional bacterial tea biological feed from development area-derived green tea residues, breeding of functional microorganisms is carried out by adoptingARTP mutagenesis and domestication screening methods, so that the bacillus pumilus BP-09 which is resistant to the high-concentration tea residues, good in biomass and good in metabolite activity is obtained. The strain can be used for tea residue fermentation, can change nutritional ingredients in the tea residues, can be applied to development of tea residue feed, and is of great significance tohigh-value utilization of tea residue resources.

Description

technical field [0001] The invention relates to the technical field of microbial mutation breeding, in particular to a bacillus pumilus BP-09 resistant to high-concentration tea dregs and its application. Background technique [0002] my country is a big tea-producing country. In 2018, the output of dry raw tea exceeded 2.6 million tons, of which green tea accounted for more than 65%. It is the main producer of green tea in the world, and its total output exceeds 75% of the total global green tea output. Green tea is a traditional beverage, and its edible part only accounts for a small part of the tea. At present, with the extension of the tea industry chain, more dry tea enters the tea deep-processing enterprises, and prepares instant tea powder and tea concentrate through high-temperature water extraction. The total amount of dry matter after deep processing of the current tea leaves is only about 3% of the dry weight of the tea leaves, and the content of active ingredient...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/36C12N15/01A23K10/12A23K10/37C12R1/07
CPCC12N1/36C12N15/01A23K10/12A23K10/37C12R2001/07C12N1/205Y02P60/87
Inventor 蒋顺进吴松刚张文黄炜乾何海峰唐谢芳
Owner 清远一生自然生物研究院有限公司
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