Strain capable of degrading collagen and application of strain

A technology of collagen and collagenase, applied in applications, animal/human protein, bacteria, etc., can solve the problems of low enzyme production activity and lack of bacterial strains, and achieve good economic benefits, increase added value, and small molecular weight.

Active Publication Date: 2021-08-06
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, people have obtained collagenase from microorganisms such as Bacillus subtilis, Bacillus cereus, actinomycetes, pseudoalteromonas, etc., but most strains have low enzyme activity or require special induction conditions to secrete high-efficiency collagen Protease strains that degrade collagen are still lacking

Method used

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  • Strain capable of degrading collagen and application of strain
  • Strain capable of degrading collagen and application of strain
  • Strain capable of degrading collagen and application of strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Screening and isolation of embodiment 1 bacterial strain

[0039] (1) Sample collection

[0040] Sediments were collected from hydrothermal vents (E 49.34627°, S 37.89947°) in the polymetallic sulphide area of ​​the Southwest Indian Ocean, placed in an ice box and brought back to the laboratory.

[0041] (2) Enrichment and domestication

[0042] Draw 500 μL of the sediment sample and add it to 50 mL of the only carbon source liquid medium, and culture it in a shaker at 25 ° C and 180 rpm for one week to obtain an enriched bacterial liquid.

[0043] (3) Screening and isolation of strains

[0044] The bacterium solution enriched in step (2) according to 10 1 , 10 2 , 10 3 , 10 4 , 10 5 Then they were inoculated on the screening medium plate respectively, cultured at 25°C for 48 hours, and further purified and inoculated on the gelatin medium by plate streaking method; then the degraded circles were formed on the gelatin medium. The strains were picked out to obtai...

Embodiment 2

[0049] The identification of embodiment 2 bacterial strain morphology

[0050] The bacterial strain screened and isolated in Example 1 was streaked onto the TYS solid medium plate, and the temperature was cultivated for 24h under the condition of 25°C, and the growth situation of the bacterial colony on the plate was observed and recorded, and the transmission electron microscope (TEM) figure of the bacterial colony morphology ,Such as figure 1 shown.

[0051] Depend on figure 1 It can be seen that the bacterial strain is on the TYS solid medium plate, and the colonies are transparent (yellow in the later stage), round, and the edges are neat, indicating that it is smooth and moist; Gram staining is red, indicating that it is a Gram-negative bacterium; transmission electron microscopy (TEM) In the figure, the cells are rod-shaped, with a length and width of 1.7-2.4×0.5-1.1 μm, single cells, and can secrete a large amount of extracellular polymers.

[0052] TYS solid medium:...

Embodiment 3

[0053] Physiological and biochemical identification of the bacterial strain of embodiment 3

[0054] The physiological and biochemical characteristics of the strains screened and isolated in Example 1 were identified by routine physiological and biochemical experiments and API 20NE and ZYM reagent strips.

[0055] The identification analysis results are shown in Table 1.

[0056] Physiological and biochemical characteristics of the bacterial strains isolated in Table 1 Example 1 and the strains of the genus Rhineheimer's close relatives

[0057]

[0058]

[0059]

[0060]

[0061] In the table: 1 is the isolated bacterial strain of embodiment 1; 2 is bacterial classification Rheinheimera tuosuensis CGMCC1.12461 T ; 3 is strain Rheinheimera longhuensis LH2-2 T ; 4 is strain Rheinheimera aestuariH29 T ; 5 is strain Rheinheimera pacifica KMM 1406 T ; 6 is strain Rheinheimera perlucidaBA131 T . + stands for positive; w stands for weak positive; -, negative reacti...

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Abstract

The invention relates to a strain capable of degrading collagen and application of the strain. The strain is preserved in China Centre for Type Culture Collection on May 8, 2021; the preservation number is CCTCC M 2021506; and the address is Wuhan University, Wuhan, China. The strain has collagen degradation capability and can be used for preparing collagenase; and the collagenase can be applied to enzymolysis of collagen and production of collagen oligopeptide. Particularly, the strain has very strong degradation capability on collagen raw materials such as ox bones, and can be used for further enzymolysis treatment on by-products of meat processing and fishery processing such as ox bones and fish skins. According to the invention, the ox bone is adopted as the raw material; the peptide fragment with the molecular weight smaller than 5000Da of the prepared collagen oligopeptide reaches 77% or above; the molecular weight is small; the uniformity is high; the collagen oligopeptide is easily absorbed and utilized by a human body; the additional value of collagen by-products is effectively improved; and the collagen oligopeptide has good economic benefits.

Description

technical field [0001] The invention relates to a bacterial strain capable of degrading collagen and its application, belonging to the technical field of biotechnology. Background technique [0002] Collagen is the most abundant protein in animals and a major structural protein in the extracellular matrix. Collagen is widely distributed and is the main component of mammalian connective tissue. It is also rich in collagen in the skin, bone, scale and other parts of aquatic and marine animals, mainly in the form of insoluble fibrous protein. These collagens are often treated as leftovers in the food processing process, which not only causes waste of biological resources, but also causes adverse effects on the environment. The full and reasonable use of these wastes in food processing can not only promote the development of animal husbandry and aquatic product processing industries, but also reduce environmental pollution. [0003] Utilizing collagen raw materials to produce ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/52C12N15/57C12P21/06C12R1/01
CPCC12N9/52C12P21/06C07K14/78
Inventor 张玉忠陈秀兰王珍张晓玉程俊慧宋晓妍
Owner SHANDONG UNIV
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