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Application of Arthrobacter nicotinovorans in hyaluronic acid degradation

A technology of Arthrobacter nicotineum and hyaluronic acid, applied in the direction of microorganism-based methods, microorganisms, biochemical equipment and methods, etc., can solve the problems that hyaluronidase has not been found in the application of hyaluronic acid degradation, etc.

Inactive Publication Date: 2010-11-17
SHANDONG UNIV AT WEIHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Arthrobacter nicotinovorans has been preserved by others in the General Microorganism Center of China Committee for the Collection of Microbial Cultures. Its preservation date is January 16, 2000, and its preservation number is CGMCC NO1.1933. It is a standard strain and its main use is In order to degrade nicotine, at present, there is no report that it can produce hyaluronidase and apply it to the degradation of hyaluronic acid

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Implementation example 1: identification of strains

[0015] The inventor of the present invention isolates a bacterial strain from hyaluronic acid spoilage liquid, and it is identified by the following method:

[0016] Bacterial chromosomal DNA was extracted, and the 16S rDNA sequence of the strain was amplified by PCR. The forward primer for amplifying 16S rDNA is 27F: 5'-AGAGTTTGATC(C / A)TGGCTCAG-3' (corresponding to the 8th-27th base position of the E.coil 16S rDNA sequence), and the reverse primer 1492R: 5 '-TACGG(C / T)TACCTTGTTACGACTT-3' (corresponding to base positions 1492-1510 of E. coli 16S rDNA).

[0017] 100μL PCR reaction system: 1×PCR buffer, 1.5mmol / L MgCl 2 , 200 μmol / L of each 4×dNTP mixture, 0.5 μmol / L of each primer, 1 μL (5 U / μL) of Taq DNA polymerase, and 2 μL of DNA stock solution. PCR reaction conditions: pre-denaturation at 96°C for 6 min; denaturation at 94°C for 1 min, renaturation at 50°C for 1 min, extension at 72°C for 2 min, 30 cycles; fin...

Embodiment 2

[0018] Implementation Example 2: Degradation of Hyaluronic Acid by Arthrobacter nicotinovorans

[0019] 1% hyaluronic acid solution (molecular weight of hyaluronic acid is 1.4 million Da), inoculated with 1% volume of Arthrobacter nicotinovorans, stirred at 30°C for 24 hours, and the molecular weight of hyaluronic acid was detected after the cultivation It is 1000 Da.

Embodiment 3

[0020] Implementation Example 3: Degradation of Hyaluronic Acid by Arthrobacter nicotinovorans

[0021] 3% hyaluronic acid solution (molecular weight of hyaluronic acid is 1.5 million Da), inoculated with 2% volume of Arthrobacter nicotinovorans, stirred at 30°C for 12 hours, and the molecular weight of hyaluronic acid was detected after the cultivation It is 60000 Da.

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PUM

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Abstract

The invention discloses a new use of Arthrobacter nicotinovorans (the preservation number is CGMCC No.1.1933 and the preservation date is January 16, 2000), and in particular relating to an application of the Arthrobacter nicotinovorans in hyaluronic acid degradation, wherein, the Arthrobacter nicotinovorans has efficient degradation effect on hyaluronic acid under normal temperature. The degradation method of the hyaluronic acid comprises two modes as follows: 1. in the later stage of fermentation production of the hyaluronic acid, adding the Arthrobacter nicotinovorans into a culture, and then carrying out mixed fermentation for 3-12 hours to obtain low molecular weight hyaluronic acid; and 2. adding the Arthrobacter nicotinovorans to a high molecular weight hyaluronic acid solution to act for 2-24 hours to obtain the low molecular weight hyaluronic acid.

Description

technical field [0001] The invention relates to the application of Arthrobacter nicotinovorans CGMCC NO1.1933, which produces hyaluronidase, in the degradation of hyaluronic acid. Background technique [0002] Hyaluronic acid, also known as hyaluronic acid, is an important physiologically active substance in the body. Hyaluronic acid has different biological activities due to its relative molecular mass in different metabolic stages. Low-molecular-weight hyaluronic acid has certain biological activities that high-molecular-weight hyaluronic acid does not have, and even the two have completely opposite effects. Low-molecular-weight hyaluronic acid is easy to absorb, and high-molecular-weight hyaluronic acid can be re-synthesized in the body to increase the content of endogenous hyaluronic acid, which can exert the effects of health care and beauty. At present, the relative molecular weight of hyaluronic acid extracted from fermentation method and animal tissue is generally m...

Claims

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Application Information

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IPC IPC(8): C12P19/00C12P19/04C12R1/06
Inventor 宋淑亮吉爱国
Owner SHANDONG UNIV AT WEIHAI