Strain for producing high-efficiency collagenase and application thereof

A collagen and strain technology, applied in the direction of animal/human proteins, microorganism-based methods, enzymes, etc., can solve the problems of lack of efficient collagenase strains, low enzyme production activity, etc., and achieve easy absorption and utilization, good economic benefits, molecular weight small effect

Active Publication Date: 2021-02-05
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, people have obtained collagenase from microorganisms such as Bacillus subtilis, Bacillus cereus, actinomycetes, and pseudoalteromonas, but most strains have low enzyme activity or require special induction conditions to produce high-efficiency collagenase strains are still lacking

Method used

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  • Strain for producing high-efficiency collagenase and application thereof
  • Strain for producing high-efficiency collagenase and application thereof
  • Strain for producing high-efficiency collagenase and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Screening and isolation of embodiment 1 bacterial strain

[0035] (1) Sample collection

[0036] P. spinosa was collected from the rocky beach in the intertidal zone (N36.22°, E120.41°) along the coast of Qingdao, placed in an ice box and brought back to the laboratory.

[0037] (2) Enrichment and domestication

[0038] Use sterilized seawater to gently wash the algae body, cut the algae body into 1 / 10TYS liquid medium with sterile scissors, and shake it for 10 minutes at 25 ° C and 180 rpm in a shaker to obtain enriched bacteria.

[0039] (3) Screening and isolation of strains

[0040] The bacterium solution enriched in step (2) according to 10 1 , 10 2 , 10 3 , 10 4 , 10 5 Stratified dilutions were carried out, and then inoculated on the screening medium plate, cultured at 25°C for 48 hours, and the strains that formed the degradation circle were picked out, and further purified by plate streaking method to obtain the highly efficient ability to degrade collag...

Embodiment 2

[0044] The identification of embodiment 2 bacterial strain morphology

[0045]Streak the bacterial strain screened and isolated in Example 1 onto a TYS solid medium plate, then invert the plate, and cultivate it at a temperature of 25°C for 24 hours, observe and record the growth of the colonies on the plate, and the atomic force microscope of the colony morphology (AFM) images and transmission electron microscope (TEM) images, such as figure 1 shown.

[0046] Depend on figure 1 It can be seen that on the TYS medium plate, the bacterial colony is white (yellow in the later stage), round, with neat edges, and the colony is wet and sticky (hard in the later stage), indicating that it is smooth and moist and opaque; Gram staining is red, indicating that It is a Gram-negative bacterium; the cells in the atomic force microscope (AFM) and transmission electron microscope (TEM) images are rod-shaped, with a length and width of 0.8-1.5×0.3-0.5 μm, single cells, polar flagella, and c...

Embodiment 3

[0047] Physiological and biochemical identification of the bacterial strain of embodiment 3

[0048] 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.

[0049] The identification analysis results are shown in Table 1.

[0050] Comparison of physiological and biochemical characteristics of the bacterial strains isolated in Table 1 Example 1 and the closely related bacterial strains of Pseudoalteromonas family

[0051]

[0052]

[0053]

[0054]

[0055]

[0056] In the table: 1 is the isolated bacterial strain of embodiment 1; 2 is bacterial classification Pseudoalteromonas mariniglutinosaKMM 3635 T ; 3 is strain Psychrosphaera aestuarii PSC101 T ; 4 is bacterial strain Psychrosphaeraaquimarina SW33 T ; 5 is the strain Psychrosphaera haliotis KDW4 T ; 6 is strain Psychrosphaerasaromensis SA4-48 T ; 7 is t...

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Abstract

The invention relates to a strain for producing high-efficiency collagenase and an application thereof. The strain is preserved in China Center for Type Culture Collection on October 9th, 2020, the preservation number is CCTCC M2020574, and the address is Wuhan University, Wuhan, China. The strain has collagen degradation capacity, and the produced collagenase can be applied to collagen degradation and collagen oligopeptide production. In particular, the strain has high degradation capability on collagen raw materials such as cattle bones, fish skin and the like and can be used for further enzymolysis treatment on meat processing and fishery processing byproducts such as the cattle bones and the fish skin, cod skin is adopted as a raw material, a peptide fragment with the molecular weightsmaller than 3000Da of the prepared collagen oligopeptide is more than 95%, the molecular weight is small, the uniformity is high, and the collagen is easy to absorb and utilize by a human body, effectively improves the added value of the collagen byproduct, and has good economic benefit.

Description

technical field [0001] The invention relates to a bacterial strain producing high-efficiency collagenase 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. It is widely distributed in connective tissues such as tendons, bones, ligaments, skin, blood vessels of mammals and fish skin and scales of aquatic animals. In tissues such as fish bones and fish bones, it mainly exists 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 c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/52C12P21/06C07K14/78C12R1/01
CPCC12N1/20C12P21/06C07K14/78C12N9/52C12Y304/24003C12R2001/01C12N1/205
Inventor 张玉忠李健程俊慧张熙颖陈秀兰宋晓妍
Owner SHANDONG UNIV
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