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Microorganism-derived algin and preparation and application thereof

A technology of alginate and microorganisms, which is applied in the field of preparation and application of alginate, can solve problems such as less research, and achieve the effect of stable chemical structure and properties

Inactive Publication Date: 2016-10-12
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, so far there are very few related studies

Method used

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  • Microorganism-derived algin and preparation and application thereof
  • Microorganism-derived algin and preparation and application thereof
  • Microorganism-derived algin and preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Screening and identification of engineering bacteria producing alginate

[0025] 1. Construction and screening of transposon library

[0026] (1) E.coli DH5α(pRK2013) (purchased from Beijing Lebo Biotech Co., Ltd.), the preparation method of the culture solution is: take the E.coli DH5α(pRK2013) bacterial solution stored in the refrigerator at -80°C, and pipette it after slightly dissolving at room temperature. Inoculate 50 μl of the bacterial solution into a test tube containing 5 ml of LB medium, add 5 μl of 15 mg / ml kanamycin aqueous solution at the same time, shake and culture at 200 rpm at 37°C overnight to obtain E.coliDH5α (pRK2013) culture medium.

[0027] E.coli DH5α(PFAC) (purchased from Beijing Loeb Biotech), the preparation method of the culture solution is as follows: take the E.coli DH5α(PFAC) bacteria solution stored in the refrigerator at -80°C, and after slightly dissolving at room temperature, use a pipette gun to transfer 50μl The bacteria...

Embodiment 2

[0061] Embodiment 2: the production of bacterial alginate

[0062] 1. Prepare culture medium

[0063] Prepare the PIA medium for producing alginate in a beaker. The medium formula is: add 45g of PIA medium powder of BD Company in 1L deionized water (composed of: peptone 20g / L, magnesium chloride 1.4g / L, potassium sulfate 10g / L, triclosan 25mg / L, agar powder 13.6g / L) and glycerin 20ml, after fully dissolving it on a magnetic stirrer, distribute to conical flasks. After autoclaving at 121°C for 20 minutes, take it out, cool to 60°C, add gentamicin at a final concentration of 200 μg / ml to the culture medium in an ultra-clean workbench, mix well and pour it into a disposable sterile petri dish, cool and solidify for later use use.

[0064] 2. Vaccination

[0065] The mutant strain of Pseudomonas aeruginosa was inoculated into the seed medium, and the shaking culture was shaken at 37°C for 24 hours to obtain the seed liquid; the final concentration of the seed medium was compos...

Embodiment 3

[0071] Example 3: Research on the interaction between different sources of alginate and human intestinal flora

[0072] 1. Preparation of culture medium

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Abstract

The invention discloses microorganism-derived algin and preparation and application thereof. The microorganism-derived algin is prepared according to a method including inoculating a pseudomonas aeruginosa mutant strain on a PIA (pseudomonas isolation agar) plate, culturing at 37 DEG C for 24-48 hours to obtain a culture containing algin, and isolating and purifying the culture so as to obtain the algin, wherein the pseudomonas aeruginosa mutant strain is constructed by inserting a transposon PFAC between a nadB gene and an algU gene of pseudomonas aeruginosa. The microorganism-derived algin has the advantages that the microorganism-derived algin is immune to alga yield, production seasons, sources and the like; compositions and characteristics of microorganism-derived algin products are easily controlled, so that chemical structures and characteristic stability of each batch of microorganism-derived algin products can be guaranteed.

Description

(1) Technical field [0001] The invention relates to the preparation and application of alginate, in particular to a method for preparing alginate by using a mutant strain of Pseudomonas aeruginosa and its application to the regulation and control of intestinal flora. (2) Background technology [0002] Alginate is an organic polysaccharide compound, which is a linear high Molecular polymer, its relative molecular weight is about 32000-200000. Because alginate has the characteristics of good thickening, stability, emulsification, film-forming, flocculation and chelating properties, and has the functions of reducing fat, removing heavy metal ions and anti-tumor, it has been widely used in food and medicine. , chemical, textile, rubber and other fields. It is precisely because of these important functions that alginate has been paid more and more attention by people all over the world. In the United States, alginate is known as "wonderful food additive"; in Japan, it is known...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/04A23L29/269A23K20/163C12R1/385
CPCC12P19/04A23V2002/00A23V2200/32
Inventor 尹业师白少峰王欣
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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