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Paenibacillus pabuli, culture medium thereof and application of paenibacillus pabuli in preparation of enteromorpha prolifera polysaccharide degrading enzymes

A technology of prolifera polysaccharide and bacillus, applied in the field of microorganisms, can solve problems such as rare reports of enzymatic degradation of prolifera polysaccharide

Active Publication Date: 2018-07-03
OCEAN UNIV OF CHINA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, enzymes with high hydrolysis efficiency are not easy to obtain, and there are few reports on the enzymatic degradation of Enteromorpha polysaccharides

Method used

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  • Paenibacillus pabuli, culture medium thereof and application of paenibacillus pabuli in preparation of enteromorpha prolifera polysaccharide degrading enzymes
  • Paenibacillus pabuli, culture medium thereof and application of paenibacillus pabuli in preparation of enteromorpha prolifera polysaccharide degrading enzymes
  • Paenibacillus pabuli, culture medium thereof and application of paenibacillus pabuli in preparation of enteromorpha prolifera polysaccharide degrading enzymes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0107] Isolation and purification of Enteromorpha polysaccharide-degrading bacteria (Paenibacillus pabuli) EP-1.

[0108] Collect sea mud from the sea area near the No. 1 bathing beach in Qingdao City, Shandong Province, add sterile seawater to dissolve it, transfer it to the primary screening medium, and shake the flask for enrichment and culture for 2-3 days. The culture solution is spread on the solid medium plate containing the enteromorpha polysaccharide, and cultivated for 3-5 days. Stain with Lugol's iodine solution, transfer the colonies with obvious degradation circles to a new Enteromorpha polysaccharide solid medium plate for culture, and repeat the streaking until a pure culture is obtained. Primary screening medium composition (mass volume ratio): Enteromorpha polysaccharide 4%, NaCl 1.5%, NaNO 3 0.5%, MgSO 4 ·7H 2 O0.05%, CaCl 2 0.01%, Iron Phosphate 0.005%, pH 7.0.

[0109] Inoculate the pure culture separated from the primary screening into the fermentatio...

Embodiment 2

[0121] The media shown in Table 1 were prepared respectively, and the preservation number was CGMCC No. 12912. Paenibacillus for feed was activated and inoculated into the media in Table 1. The inoculum size is 5%, the culture temperature is 28°C, the shaker speed is 200rpm, cultured for 24h, and the OD of the bacterial solution is measured 600 value.

[0122] The obtained bacterial solution was centrifuged at 6000 rpm for 20 minutes to obtain a fermentation supernatant; at 4°C, the obtained fermentation supernatant was sterilized through a 0.22 μm filter membrane, concentrated by ultrafiltration, desalted by dialysis, and vacuum freeze-dried to obtain Enteromorpha polysaccharide degrading enzyme. Determination of enzyme activity. The detection method of enzyme activity is DNS method, and the definition of enzyme activity: 1mL enzyme solution produces 1μg reducing sugar in 1min as an activity unit.

[0123] Table 1, medium formula

[0124]

[0125]

[0126]

[0127...

Embodiment 3

[0129] Adopt the medium with the best effect in Table 1 (the formula is Enteromorpha polysaccharide 0.2%, peptone 0.6%, yeast extract powder 0.7%, NaCl 0.5%, MgSO 4 ·7H 2 O 0.1%, Na 2 HP0 4 6mmol / L) to prepare Enteromorpha polysaccharide degrading enzyme by fermentation. Pre-treated and activated before fermentation.

[0130] 1) Pre-treatment: Streak Paenibacillus for feed with the preservation number CGMCC NO.12912 on the solid medium of Enteromorpha polysaccharide, and culture it at 25-35°C for 5-7 days;

[0131] 2) Activation: Inoculate 1 to 3 loops of the solid culture medium strain into a test tube of seed culture solution, and culture at 25 to 35° C. at 150 to 210 rpm for 12 to 24 hours.

[0132] 3) Fermentation: insert the seed culture medium into the fermentation culture medium, culture on a shaker for 24-48 hours, and obtain the fermented product.

[0133] Step 3 carries out the fermentation test of different parameters, and each parameter is as table 2:

[0134...

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Abstract

The invention relates to the technical field of microbes and in particular relates to paenibacillus pabuli, a culture medium thereof and application of paenibacillus pabuli in preparation of enteromorpha prolifera polysaccharide degrading enzymes. The paenibacillus pabuli provided by the invention has a collection number of CGMCC NO.12912. The invention further provides a culture medium for screening culture, a culture medium for activating culture and a culture medium for fermentation. The invention also provides a method for preparing enteromorpha prolifera polysaccharide degrading enzymes by using culture fermentation and enteromorpha prolifera polysaccharide degrading enzymes prepared by fermentation. The enteromorpha prolifera polysaccharide degrading enzymes produced by the culture provided by the invention and fermentation have excellent enzymatic activities and are capable of degrading enteromorpha prolifera polysaccharides, and enteromorpha prolifera polysaccharide oligosaccharide of different degrees of polymerization can be prepared according to different parameters. Experiments prove that the activity of the enteromorpha prolifera polysaccharide degrading enzyme provided by the invention can reach 1.03U / mL.

Description

technical field [0001] The invention relates to the technical field of microbes, in particular to Paenibacillus for feed, its culture medium and its application in the preparation of enteromorpha polysaccharide degrading enzyme. Background technique [0002] Seaweed sulfated polysaccharide is a sulfate-based heteropolysaccharide extracted from seaweed. Its structure contains sulfate groups, and its monosaccharide composition includes glucose (Glc), rhamnose (Rha), xylose (Xyl), and glucuronic acid (GlcA). , mannose (Man) and galactose (Gal), etc., the sulfate group is mainly connected to rhamnose, but it varies with the type of Enteromorpha and the extraction method. [0003] Existing studies have confirmed that Enteromorpha polysaccharides have various physiological activities such as immune regulation, anti-tumor, anti-coagulation, anti-virus, anti-oxidation, and hypolipidemic. Moreover, Enteromorpha polysaccharide has excellent thermoreversible gelation, hydrophilic and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02C12N9/24C12R1/01
CPCC12N1/02C12N1/20C12N9/2402C12N1/205C12R2001/01
Inventor 刘晨光于钰丁松李嘉欣孙茜
Owner OCEAN UNIV OF CHINA
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