Bacillus velezensis and application thereof

A Bacillus and enrichment culture technology, applied in the field of microorganisms, can solve the problems of many by-products, environmental pollution, and high saponification costs, and achieve the effects of high enzyme production efficiency, fast fermentation process, and efficient degradation.

Active Publication Date: 2020-10-16
CHINA AGRI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The composition of astaxanthin ester is complex, it is not easy to be absorbed and utilized in the body, and it needs to be hydrolyzed to obtain astaxanthin monomer
The most commonly used method for the conversion of astaxanthin esters is the alkali saponification method, but astaxanthin monomers and esters are unstable under alkaline conditions, astaxanthin monomers are easily oxidized, and saponification costs are high, with many by-products, It is difficult to recover astaxanthin monomer and seriously pollutes the environment, so it is not conducive to industrial production; the use of biocatalyst-lipase not only has high conversion efficiency, but also is pollution-free

Method used

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  • Bacillus velezensis and application thereof
  • Bacillus velezensis and application thereof
  • Bacillus velezensis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Screening of high-yield strains of astaxanthin esterase

[0056] Weigh 1g of soil sample, add it to 10mL 0.9% sterile normal saline, fully shake it under the condition of 200r / min, take 1mL supernatant in 50mL enrichment medium after standing still, keep constant temperature at 37℃ and 200r / min Shake culture for 24 hours, transfer 1 mL to a new enrichment medium for the second enrichment, and cultivate for 24 hours under the same conditions for the third enrichment to obtain the enriched bacterial liquid.

[0057] The enrichment medium includes the following components: 0.02% of sucrose, 0.02% of yeast extract powder, 0.05% of sodium chloride, 0.35% of disodium hydrogen phosphate, 0.15% of ammonium sulfate, 0.15% of dipotassium hydrogen phosphate, and 0.05% of anhydrous magnesium sulfate %, olive oil emulsion 1%.

[0058] Dilute the enriched bacterial solution to 10 -6 Double the concentration, the bacteria solution is evenly spread on the Rhodamine B selection medium...

Embodiment 2

[0064] Enzyme Activity Measurement of High-Producing Astaxanthin Esterase Strain

[0065] The method used in the measurement of enzyme activity in this embodiment is titration.

[0066] Olive oil emulsion was used as the substrate for the reaction. Add 50mL phosphate buffer solution and 40mL olive oil emulsion to the reaction system, mix well and react in a water bath at 40°C for 5 minutes, then take it out, add 10mL centrifuged fermentation supernatant to the reaction system, mix well and react in a water bath at 40°C Take it out after 15 minutes, add 15mL of 95% absolute ethanol to terminate the reaction, add two drops of 1% phenolphthalein indicator, and titrate with 0.05mmol / L sodium hydroxide standard solution until the end point of the titration is reddish and does not fade for 30s, record the consumption The volume of 0.05mmol / L sodium hydroxide standard solution.

[0067] x=(Vt–V0) N / Vm t

[0068] In the formula, x is the enzyme activity of the sample, in U / mL; Vt i...

Embodiment 3

[0070] Morphological identification of a strain with high astaxanthin esterase production

[0071] A small amount of the bacterial strains screened in Example 1 were fermented and cultured in the fermentation medium, cultured at 37°C for 24 hours, diluted, spread on a solid plate, cultured at 37°C, observed single colony shape, size, edge, surface, transparency, etc., and Gram staining was carried out, and the morphological characteristics of the strains were observed under an optical microscope.

[0072] Morphological characteristics of colonies: round and neat, smooth and moist colonies, slightly raised. The bacteria are rod-shaped, Gram staining is positive, spores are formed, and they do not expand. Gram stain see figure 1 , spore staining see figure 2 .

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Abstract

The invention relates to the technical field of microorganisms, in particular to bacillus velezensis and an application thereof. The preservation number of the bacillus velezensis Lpl-wx is CGMCC No.17045. The bacterial strain can be applied to preparation of astaxanthin esterases. The bacillus velezensis belongs to prokaryotes, so that the fermentation process is quick, enzyme production efficiency is high, and the astaxanthin esterases can be quickly and efficiently obtained. The bacterial strain can be further applied to preparation of an astaxanthin single body and can effectively degradeastaxanthin esters, the astaxanthin single body can be obtained, a green efficient catalyzing way few in by-products is provided, and a technical basis is provided for preparation and promotion of theastaxanthin single body.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a bacillus velei and its application. Background technique [0002] Astaxanthin (also known as 3,3’-dihydroxy-4,4’-diketone-β,β-carotene) is an oxygen-containing derivative of carotene, with the structural formula C 40 h 52 o 4 , Molecular weight 596.86, molecular structure contains very long conjugated double bond and α-hydroxy ketone, widely exists in organisms, especially aquatic organisms such as shrimp, crab, algae and fish. Astaxanthin extracted from Haematococcus pluvialis mainly exists in three states: free astaxanthin monomer accounts for only 5%, astaxanthin monoester accounts for 70%, astaxanthin diester accounts for 25%, and in the extract The fatty acid composition is mainly in the form of long-chain saturated and unsaturated fatty acids, including C16:0 (7%), C18:0 (7%), C19:0 (7%), C20:0 (7%) and C18:1 (56%), mainly oleic acid, which is similar in struct...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02C12N9/16C12P23/00C12R1/07
CPCC12N1/20C12N1/02C12N9/16C12P23/00C12R2001/07C12N1/205
Inventor 李平兰汪伯良刘力王瑶谢清张莹武瑞赟
Owner CHINA AGRI UNIV
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