Arthrobacter sp. PL-410 and application thereof in producing chondroitin lyase

A technology of PL-410 and Arthrobacter, which is applied in the field of microorganisms and achieves the effect of broad application prospects

Pending Publication Date: 2022-05-13
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few high-activity chondroitin lyases with application value at present, so it is of great significance to screen new chondroitin lyase-producing bacteria, and to isolate and identify new stable and high-activity chondroitin lyases

Method used

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  • Arthrobacter sp. PL-410 and application thereof in producing chondroitin lyase
  • Arthrobacter sp. PL-410 and application thereof in producing chondroitin lyase
  • Arthrobacter sp. PL-410 and application thereof in producing chondroitin lyase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 Acquisition of Pseudarthrobacter sp. of the present invention

[0049] Take a little soil into a conical flask, add 9 mL of sterile saline, shake fully, draw 1 mL of the supernatant into the enriched medium, and shake at 30°C and 200 rpm for 18 hours to obtain an enriched bacterial solution. Take 1mL of enriched bacterial solution and dilute to 10 -6 Concentration, take a suitable concentration gradient of bacterial liquid evenly spread on the separation medium, cultivate at 30 °C for 3 days, pick colonies of different shapes and carry out streak purification to obtain a single colony. A single colony was planted on the primary screening medium, and after culturing at 30°C for 3 days, 5 mL of 2 mol / L acetic acid solution was added to the primary screening medium, and immersed for 5 minutes to observe whether a transparent circle was produced. Rescreen. The strains with transparent circles were inoculated into the re-screening medium respectively, cultured wi...

Embodiment 2

[0069] The cultivation of embodiment 2 pseudoarthrobacter of the present invention

[0070] The pseudoarthrobacter PL-410 strain was picked and inoculated into the liquid medium, and the seed solution was prepared by shaking culture at 26° C. and 200 rpm overnight. Draw the seed liquid, inoculate it in the liquid medium at a volume percentage of 1%, 26 ° C, 200 rpm shaking culture, the growth curve is as follows image 3 As shown, the strain entered the logarithmic growth phase after 6h, the stable phase after 28-44h, and the decay phase after 44h.

[0071] The components per liter of the above liquid medium are as follows:

[0072] Tryptone 10g, yeast extract 5g, NaCl 10g, the balance is deionized water, pH value is 7.2.

Embodiment 3

[0073] Example 3 Whole Genome Sequencing and Sequence Analysis of Pseudomonas Pseudomonas of the Present Invention

[0074] According to the culture method described in Example 2, the PL-410 strain was cultured for 18 hours, 1 L of the bacterial solution was taken, centrifuged at 10,000 rpm for 10 minutes, and the bacterial pellet was collected. Sequencing. The BLAST (Basic Local Alignment Search Tool) of the NCBI (National Center for Biotechnology Information) website was used to analyze the sequencing results. The NCBI analysis results showed that the Pseudomonas PL-410 genome carries a chondroitin lyase gene 1410, the encoding of the gene 1410. The length of the region is 2397bp, and the nucleotide sequence is shown in SEQ ID NO.2. The chondroitin lyase L410 encoded by the gene 1410 contains 798 amino acids in total, and its amino acid sequence is shown in SEQ ID NO.3.

[0075] Using the BLASTp online analysis of the NCBI website, the results show that the most similar am...

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Abstract

The invention relates to the technical field of microorganisms, in particular to arthrobacter pseudoarthrobacter PL-410 and application of the arthrobacter pseudoarthrobacter PL-410 in production of chondroitin lyase. The preservation number of the pseudoarthrobacter sp. PL-410 provided by the invention is CGMCC (China General Microbiological Culture Collection Center) No.22736, and the preservation number of the pseudoarthrobacter sp. PL-410 is CGMCC No.22736. The pseudoarthrobacter sp. PL-410 disclosed by the invention can be used for preparing chondroitin lyase (AC type), and the chondroitin lyase can be used for researching the structure of chondroitin sulfate and constructing a chondroitin sulfate oligosaccharide library, can also be applied to the field of medicines, and has a wide application prospect.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a pseudoarthrobacter PL-410 and its application in producing chondroitin lyase. Background technique [0002] Chondroitin sulfate (CS) is a class of glycosaminoglycans extracted from animal tissues, widely present in the cell surface and extracellular matrix of vertebrate connective tissues, and has a variety of biological activities, such as repairing articular cartilage. , anti-oxidation, anti-aging, anti-coagulation, anti-thrombosis, hypolipidemic, anti-atherosclerosis and anti-tumor, etc., are widely used in medicine, food and cosmetics industries. Common CSs have high molecular weight and high charge density, resulting in low bioavailability and limiting their biological efficacy. More and more evidences show that low-molecular-weight CS has the advantages of low viscosity, good solubility, and easy absorption, which can effectively overcome the shortcomings of low b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/88C40B50/06C12P19/12C12P19/04C12P19/00A61K35/74A61P19/00A61P19/08C12R1/01
CPCC12N1/20C12N9/88C40B50/06C12P19/12C12P19/04C12P19/00C12Y402/02005A61K35/74A61P19/00A61P19/08
Inventor 尚楠肖梦圆武瑞赟李平兰
Owner CHINA AGRI UNIV
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