Raueria ornithinolyticum g10, enzyme production method, product and application
A technology of ornithine and enzyme-producing medium, which is applied in the field of microorganisms, can solve the problems of long enzyme-producing fermentation time, high fermentation temperature, and high cost of enzyme-producing medium, and achieve short cultivation time, simple ingredients, and wide application potential Effect
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0034] Example 1, Raoultella ornithinolytica G10 (hereinafter referred to as Raoultella ornithinolytica G10 or strain G10), its biological deposit number is CCTCC NO: M 2019485. The isolation of Raoulia ornithine solution G10, the steps are as follows:
[0035] The sea mud samples were collected in the low tide area of Haiyifang Park in Lianyungang, diluted with sterile distilled water, spread on the screening medium plate, cultured at 25°C for 48-72 hours, and colonies with transparent circles were selected. The colonies were picked and inoculated into the enzyme-producing medium, cultured at 25°C for 24 hours, and the culture was centrifuged at 8000 rpm for 3 minutes to obtain the supernatant, and the chitin deacetylase activity in it was detected. In a 10ml centrifuge tube, add 3ml of 0.05mol / L phosphate buffer, add 1ml of fermentation supernatant, and then add 1ml of 0.2g / L p-nitroacetanilide solution, and place it in a 30°C water bath for enzymatic reaction 15min, afte...
Embodiment 2
[0057] Embodiment 2, the method for producing chitin deacetylase by Raoulia ornithine solubilization G10 described in embodiment 1, the steps are as follows:
[0058] (1) Preparation of seed liquid: inoculate the slanted surface of G10 strain into the seed medium, rotate speed 180rpm, fill 20% of liquid, and cultivate at 30°C for 12h to obtain seed liquid; wherein the seed medium is kelp juice, and the preparation method is: dry The kelp was ground into 80-mesh powder, weighed 20g and added to 1000ml tap water, boiled for 20min, supplemented the water loss, and filtered.
[0059] (2) Enzyme production culture: inoculate the seed liquid in the enzyme production medium with 1% (v / v) inoculation amount, 200rpm, 30% liquid volume, culture at 25°C for 24h, centrifuge or filter the cells, and collect the supernatant , that is, the chitin deacetylase fermentation broth is obtained. According to the method in Example 1, the enzyme activity of the fermentation broth was determined to ...
Embodiment 3
[0070] Embodiment 3, utilizes ornithine solubilization Raoulia G10 to hydrolyze mannan, the steps are as follows:
[0071] The mannanase enzyme production medium was inoculated with Raoulia ornithine solubilization G10, cultured at 180 rpm, 40% liquid volume, and 32 °C for 34 h, the cells were centrifuged to precipitate, and the supernatant was collected to obtain the mannanase extraction liquid. Take 100ml of 0.05mol / L phosphate buffer solution (pH 7.0), add 0.4g of konjac powder, heat and stir to dissolve with a glass rod, and prepare a substrate solution. Take 0.9ml of the substrate solution and put it in a 10ml centrifuge tube, add 0.1ml of the enzyme extract, shake well, put it in a 55°C water bath, and carry out the enzymatic reaction for 10min. After the reaction was completed, 2 ml of DNS solution was added, and the mixture was placed in an induction cooker and reacted in boiling water for 5 min. The absorbance value at 540nm was measured with a spectrophotometer. T...
PUM
| Property | Measurement | Unit |
|---|---|---|
| diameter | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More