Photobacterium sp. QA16 as well as culture method and application thereof

A technology of photobacteria and QA16, which is applied in the field of microorganisms, can solve problems such as sources, and achieve the effect of broad application prospects

Active Publication Date: 2020-12-25
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, currently there are few highly active CS / DS degrading enzymes with application value and most of them come from land, and there are even fewer CS / DS degrading enzymes from marine sources. Therefore, it is of great significance to find new CS / DS degrading bacteria from marine sources.

Method used

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  • Photobacterium sp. QA16 as well as culture method and application thereof
  • Photobacterium sp. QA16 as well as culture method and application thereof
  • Photobacterium sp. QA16 as well as culture method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1, the acquisition of Photobacterium sp. QA16

[0042] Take the sea mud leaching solution, add 1 mL of the supernatant to 9 mL of sterile water, and dilute to 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 Concentration gradient, then inoculate and spread on the only carbon source solid medium by conventional dilution method or streaking method, culture at 30°C for 1 day, count the colonies, then select the colonies with obvious differences in colony morphology, and repeat the streaking Inoculate on the corresponding full nutrient agar plate until a single colony is obtained after purification, and then transfer to the corresponding agar slant for later use.

[0043] The cultured strains were inoculated on the only carbon source liquid medium for cultivation. Cultivate at 200 rpm and 30°C for 72 hours, observe the turbidity of the bacterial solution, and take the culture supernatant for carbazole reaction to detect the consumption of carbon sources. The enzy...

Embodiment 2

[0048] Embodiment 2, the cultivation of Photobacterium QA16 bacterium liquid

[0049] The cultivation method of Photobacterium QA16 bacterium liquid, the steps are as follows:

[0050] (1) Inoculate the Photobacterium QA16 strain into the liquid medium, and culture it on a shaking table for 10 hours at a temperature of 28-30° C. and a rotation speed of 200 rpm to obtain a seed solution;

[0051] (2) Get the seed liquid that step (1) makes, inoculate in the liquid culture medium by 7% volume percentage, be 25~30 ℃ in temperature, under the condition that dissolved oxygen (being dissolved oxygen saturation) is 30% , expanded culture for 2 hours, and obtained Photobacterium QA16 bacteria liquid.

[0052] The components per liter of the above-mentioned liquid culture medium are as follows:

[0053] K 2 HPO 4 1g, MgSO 4 ·7H 2 O 0.5g, NaCl 30g, CaCl 2 2H 2 O 0.1g, FeSO 4 0.001g, NH 4 Cl1g, tryptone 10g, yeast extract 5g, water 1000mL, pH value is 7.0.

Embodiment 3

[0054] Example 3, Application of Photobacterium QA16 in the Preparation of Chondroitin Sulfate Lyase

[0055] The application of Photobacterium QA16 in the preparation of chondroitin sulfate lyase, the steps are as follows:

[0056] (1) Inoculate the Photobacterium QA16 strain into a liquid medium, and culture it on a shaking table for 16 hours at a temperature of 28-30° C. and a rotation speed of 300 rpm to obtain a seed solution;

[0057] (2) Get the seed liquid that step (1) makes, inoculate in liquid culture medium by 8% volume percentage, be 25~30 ℃ at temperature, under the condition that dissolved oxygen (being dissolved oxygen saturation) is 25% , expand the culture for 5 hours, add chondroitin sulfate, so that the mass concentration of chondroitin sulfate is 0.01-0.02%, continue to cultivate for 72 hours, and obtain the photobacterium QA16 fermentation liquid;

[0058] (3) Take the photobacterium QA16 fermented liquid obtained in step (3), centrifuge at 12,000rpm for...

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Abstract

The invention relates to a photobacterium sp. QA16 as well as a culture method and application thereof. The photobacterium sp. QA16 is preserved in the China General Microbiological Culture CollectionCenter (CGMCC) on August 26, 2020, the preservation address is Institute of Microbiology, Chinese Academy of Sciences, No.3, Yard 1, West Beichen Road, Chaoyang District, Beijing, and the preservation number is CGMCC NO.20556. The photobacterium sp. QA16 strain can be used for preparing a chondroitin sulfate lyase, and the chondroitin sulfate lyase can degrade chondroitin sulfate and dermatan sulfate and is chondroitin sulfate ABC enzyme. The photobacterium sp. QA16 strain can be applied to structural research of the chondroitin sulfate, can also be applied to the fields of medicines, cosmetics and the like, and has wide application prospects.

Description

technical field [0001] The invention relates to a photobacterium QA16 and its cultivation method and application, belonging to the technical field of microbes. Background technique [0002] Chondroitin sulfate (CS) is an acidic linear anionic polysaccharide composed of disaccharide units composed of D-glucuronic acid and N-acetylgalactosamine linked repeatedly through β-1,4 glycosidic bonds. It is an important class of sugars Aminoglycans are also typical representatives of complex structures in glycosaminoglycans. CS sugar chains generally contain 50-70 disaccharide units, and the molecular weight is between 5000-50000Da. CS often exists widely on the surface of animal cells and in the extracellular matrix. A large number of studies have shown that CS has a variety of biological functions and can participate in a series of physiological and pathological processes in the body. During the synthesis of CS, under the action of various glycosyl modification enzymes (sulfatylt...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/88C12N15/60C12P19/04C12P19/00C12R1/01
CPCC12N1/20C12N9/88C12Y402/99006C12P19/04C12P19/00C12R2001/01C12N1/205
Inventor 李福川张庆冬路丹荣魏琳
Owner SHANDONG UNIV
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