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Fermentation production method and application of high-yield exopolysaccharide strain and amylase thereof

A technology of extracellular polysaccharides and production methods, applied in the field of polysaccharides, to achieve the effects of enhancing immunity and low production costs

Active Publication Date: 2013-03-20
厦门百拓生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the extraction of polysaccharides by fermentation of this strain at home and abroad so far.

Method used

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  • Fermentation production method and application of high-yield exopolysaccharide strain and amylase thereof
  • Fermentation production method and application of high-yield exopolysaccharide strain and amylase thereof
  • Fermentation production method and application of high-yield exopolysaccharide strain and amylase thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Identification of a high-production exopolysaccharide strain

[0031] Classical microbial biochemical tests show that the obtained high-yield exopolysaccharide strain has the characteristics of Proteus, a kind of Proteus penninii, Gram staining is negative, and the size is (0.4~0.6)μm×(1~3) μm, has obvious pleomorphism, is filamentous, has complete cell wall, no capsule, and there are flagella all over the body in young cultures; it grows diffusely on solid medium, forming alternating thickness with the inoculation site as the center, Concentric circular layers of wavy lawns. see figure 1 with 2 .

[0032] Biochemical profile:

[0033] This strain does not ferment lactose, but can produce urease, which decomposes urea and releases ammonia. Indole negative, oxidase negative. Catalase test was positive. Can break down collagen. It can ferment glucose to produce gas, but does not ferment lactose, citric acid and malonic acid. Methyl red staining is posi...

Embodiment 2

[0034] Example 2: A production process for the production of exopolysaccharides by fermentation of exopolysaccharide strains

[0035] 1) culture medium

[0036] Take the preparation of 1L medium stock solution as the seed solution, the inoculum size is 1:50, and the seed tank is 50L as an example.

[0037] Recipe: peptone 10g, KH 2 PO 4 5g, inorganic salt 25ml (MgSO 4 0.5g / L, MnSO 4 0.18g / L, FeSO 4 7H 2 (00.1g / L).

[0038] After preparation, use NaOH solution to adjust the pH to 7.2-7.4, and then pack it separately.

[0039] 2) Sterilization

[0040] Put the aliquoted medium stock solution into a high-pressure steam sterilizer for sterilization. The sterilization temperature is 121°C and the time is 20 minutes.

[0041] 3) Vaccination

[0042] After cooling the sterilized medium stock solution, inoculate the bacteria on the aseptic operation table, and pay attention to prevent bacterial contamination during the operation.

[0043] 4) Shaker culture

[0044] After ...

Embodiment 3

[0059] Example 3: A method for measuring the content of exopolysaccharide produced by the exopolysaccharide strain

[0060] 1) Reagent

[0061] Water is distilled water. Concentrated sulfuric acid (GR), 95% ethanol and anhydrous glucose are commercially available analytically pure.

[0062] Phenol reagent: Take 200g of phenol (CP), add 0.2g of aluminum flakes and 0.1g of sodium bicarbonate, distill and collect the distillate at about 180°C. Make freshly distilled phenol into a solution with a concentration of 50g / L, put it into a brown bottle, and store it in the refrigerator for later use.

[0063] 2) Instrument

[0064] 722 spectrophotometer, BECKMAN low temperature refrigerated centrifuge, vacuum freeze dryer, constant temperature box, rotary evaporator.

[0065] 3) Operation method

[0066] 3.1 Determination of glucose standard curve: Accurately weigh 20 mg of standard glucose dried to constant weight at 105°C and dissolve in 100 ml of double-distilled water respectiv...

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Abstract

The invention relates to an amylase, in particular to a fermentation production method and an application of a high-yield exopolysaccharide strain and an amylase thereof. The strain is a proteus penneri PZ-1. The method comprises the following steps: inoculating a culture medium with the roteus penneri PZ-1; fermenting the culture medium to obtain fermentation culture fluid; cooling and centrifuging the fermentation culture fluid to remove a strain body; getting supernatant fluid; precipitating the supernatant fluid with industrial alcohol; standing still precipitate liquid is obtained; centrifuging the precipitate liquid and taking out a precipitate to obtain crude amylase; and purifying the crude amylase to obtain exopolysaccharide. Proved by a series of experiments, the strain has the proteus characteristics, is used for producing the exopolysaccharide with high yield, and has obvious effects on improving immunity and capability to resist diseases; and the exopolysaccharide producedby fermentation can be used for preparing veterinary injecta, veterinary powder, feed additives, an immunologic adjuvant and functional raw material of healthcare foods.

Description

technical field [0001] The invention relates to a polysaccharide, in particular to a bacterial strain and a fermentative production method and application of the exopolysaccharide. Background technique [0002] Regarding the research on the fermentation of polysaccharides of P. pennellii, from the articles published at home and abroad, there are only articles on the isolation of Proteus pennzii in China, but no related studies; there are a large number of articles abroad to study the polysaccharide components of Proteus pennikei and its components. Antigenicity, from different serotypes such as Proteus penneril, 2, 3, 8, 10, 14, 15, 16, 18, 19, 20, 22, 25, 26, 31, 32, 35, 41, 42, 45, 52, 60, 62, 63, 71, 75, 103, etc. ([1] F.W.Hichman, A.G.Steigerwalt, J.J.Farmer III, And Don J.Brenner. Identification of Proteus penneri sp.nov., Formerly Known As Proteus vulgaris Indole Negative orAs Proteus vulgaris Biogroup 1, J Of Clinical Microbiology, 1982, 15(6):1097-1102), by Kondakov...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P19/04A61K31/715A61K39/39A61K9/08A61K9/14A61P37/04A23K1/16A23L1/29C12R1/37A23K20/163A23L33/00
Inventor 谢荔朋彭宣宪
Owner 厦门百拓生物工程有限公司
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