Arthrobacter, uses for produced xanthine oxidase and production method thereof

A technology of xanthine oxidase and Arthrobacter, which is applied in the field of microorganisms, can solve the problems of unstable content and quality of xanthine oxidase, limit the expansion of application fields, and high production costs, and achieve short production cycle, easy operation, and product activity high effect

Inactive Publication Date: 2009-04-08
JIMEI UNIV
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  • Claims
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Problems solved by technology

The xanthine oxidase used now has a single source, mainly isolated and extracted from milk butter
Raw milk is affected by different origins and different types of

Method used

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  • Arthrobacter, uses for produced xanthine oxidase and production method thereof

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preparation example Construction

[0034] The preparation method that Arthrobacter sp.XL2006 is applied to ferment and produce xanthine oxidase comprises the following steps:

[0035](1) The strain Arthrobacter sp.XL2006 was inoculated from a slant into a liquid seed medium with a liquid content of 20% for shaking shake flask culture. The shaker speed was 200 rpm, the temperature was 30 °C, and the culture time was 20 h. A grade seed liquid;

[0036] (2) Transfer the primary seed liquid into the enzyme-producing fermentation medium according to the inoculation amount of 5-10% of the fermentation liquid volume, and cultivate at 32° C. for 42 hours to reach the peak enzyme production;

[0037] (3) Centrifuge at 5000-8000rpm to collect (2) Arthrobacter sp.XL2006 cells in the fermentation broth, use ultrasonic method and lysozyme method to break the cell wall, and centrifuge at 8000-10000rpm to remove cell debris, and the collected supernatant is the crude enzyme solution ;

[0038] (4) The crude enzyme solution ...

Embodiment

[0050] Soil samples rich in nucleosides were collected, and xanthine and a small amount of yeast powder were used as carbon sources, nitrogen sources and energy sources for microbial growth. Single colony isolation was performed on medium agar plates. Then a single colony was picked for liquid fermentation to determine the enzyme activity and re-screened to obtain a strain of bacteria with the characteristic of high xanthine oxidase production. The bacterium was identified as a new species of Arthrobacter through morphological observation, physiological and biochemical analysis and 16S rRNA sequence. (The selection medium is: yeast extract 2.0g, glucose 5g, NaCl 5.0g, agar 15g, H 2 o 2 1L, pH8.0)

[0051] The specific steps for the isolation and purification of Arthrobacter sp.XL2006 are as follows: collect 20 soil samples of different vegetation in many places, take a small amount of samples and inoculate them in 100mL selection medium, and carry out constant temperature ...

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Abstract

The invention provides an Arthrobacter and an application of the Arthrobacter for generating xanthine oxidase and a preparation method thereof. The Arthrobacter XL2006 derives from nature and the16S rRNA sequence of the Arthrobacter XL2006 is shown in a sequence table SEQ ID NO.1. The Arthrobacter XL2006 is applied to producing, extracting and purifying the xanthine oxidase and preparing a diagnostic kit. The Arthrobacter XL2006 is characterized by the production of the xanthine oxidase and can adopt hypoxanthine or xanthine as a unique source of carbon and nitrogen and a unique energy source. The fermentation and the production of the xanthine oxidase are characterized by simple nutritional requirements, easy culture and short fermentation time; the produced xanthine oxidase has quite high activity, and strains provided by the invention are utilized to ferment and extract the xanthine oxidase and the prepared diagnostic kit can be applied to clinically testing the levels of hypoxanthine and xanthine in blood serum of patients.

Description

technical field [0001] The invention belongs to the technical field of microbes, and more specifically relates to an Arthrobacter sp. XL2006 and its application and preparation method for producing xanthine oxidase. technical background [0002] Xanthine oxidase (Xanthine oxidase, XOD, EC 1.2.3.22) is the rate-limiting enzyme of purine nucleoside metabolic pathway. It catalyzes the oxidation of hypoxanthine and xanthine to uric acid, which is closely related to ischemic reperfusion injury, gout, hyperuricemia, rheumatoid arthritis, myocardial infarction, hepatitis and other diseases. According to information and literature search, xanthine oxidase can be developed into an enzyme for medical testing, which can be used to detect serum inorganic phosphorus, serum superoxide dismutase activity, extracellular hypoxanthine and xanthine levels after cell damage, and Inosine (inosine), guanine, guanosine, guaninase and nucleoside phosphorylase activities, etc. The xanthine oxidase...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/02C12R1/06
Inventor 李忠琴关瑞章王武杨海麟
Owner JIMEI UNIV
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