Preparation method and application of enteric bacilli Z0206 enriched-selenium polysaccharide

An enterobacteriaceae and selenium-enriched technology, applied in the field of microbiology and bioengineering, can solve the problems of high toxicity and low bioavailability of inorganic selenium compounds, and achieve significant GSH-Px activity and MDA content, GSH-Px activity and MDA content. The effect of content regulation and high product purity

Inactive Publication Date: 2014-10-08
ZHEJIANG UNIV
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The toxicity of inorganic selenium compounds is much higher than that of organic selenium compounds, and their bioavailability is low

Method used

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  • Preparation method and application of enteric bacilli Z0206 enriched-selenium polysaccharide
  • Preparation method and application of enteric bacilli Z0206 enriched-selenium polysaccharide
  • Preparation method and application of enteric bacilli Z0206 enriched-selenium polysaccharide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1, the preparation method of Enterobacter Z0206 selenium-enriched polysaccharide

[0022] The formula of PDA enriched medium is as follows: 20% potato, 0.3% peptone, 0.3% yeast extract, 2% glucose and 1.6% agar powder. The formula of PDA enriched liquid medium is as follows: 20% potato, 0.3% peptone, 0.3% yeast extract, 2% glucose; the formula of fermentation medium is as follows: 20% cornstarch, 1.6% soybean meal powder, 0.6% dry corn steep liquor powder, 2 HPO 4 .3H 2 O0.3%, KH 2 PO 4 0.3%, CaCl 2 0.1%.

[0023] (1) Acclimatization of Enterobacter Z0206 tolerance to selenium:

[0024]Pick selenium-tolerant strains, spread them on the PDA-enriched medium plates with selenium content of 10, 20, 30, 40, and 50 μg / mL in turn, and carry out selenium concentration gradient acclimatization. Each gradient was incubated at 30°C for 36h. Finally, the well-grown colonies were picked from the PDA enriched medium plate with a selenium content of 50 μg / mL, streak...

Embodiment 2

[0042] Example 2, Application of phenol-sulfuric acid method to detect polysaccharide content in Enterobacter Z0206 selenium-enriched polysaccharide

[0043] The method is as follows: Accurately weigh 20 mg of standard glucose in a 500ml volumetric flask, add water to the mark, absorb 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml, 1.2ml, 1.4ml, 1.6ml, 1.8ml and add water to make up to 2.0ml, then add 1.0ml of 6.0% phenol and 5.0ml of concentrated sulfuric acid, let it stand for 10min, shake well, bathe in water for 30min, measure the optical density at 490nm after 20min, use 2.0ml of water as blank, and the abscissa is the number of micrograms of polysaccharide , and the ordinate is the optical density value, and a standard curve is drawn. Draw 1.0ml of the sample solution (equivalent to about 40 μg of selenium-enriched polysaccharide), operate according to the above steps, measure the optical density, and calculate the polysaccharide content with the standard curve.

Embodiment 3

[0044] Example 3. Determination of protein content in Enterobacter Z0206 selenium-enriched polysaccharide by Kjeldahl method.

[0045] The Kjeldahl method (GB / T5009.5-1985) stipulated by the national standard was used to determine the protein content in the selenium-enriched polysaccharide of Enterobacter Z0206.

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Abstract

The invention discloses a preparation method of an enteric bacilli Z0206 enriched-selenium polysaccharide. The method comprises the following steps: selecting an enteric bacilli Z0206 selenium-resistant strain, coating the selenium-resistant strain on PDA enriched medium containing selenium to carry out domestication, and determining the optimal selenium feeding time and cultivation time by determining a growth curve of bacteria; inoculating the domesticated enteric bacilli Z0206 strain into the PDA enriched liquid medium to carry out shake-flask fermentation, thereby obtaining a fermentation liquor containing the enteric bacilli Z0206 enriched-selenium polysaccharide; collecting supernate from the fermentation liquor, thereby obtaining enriched-selenium crude polysaccharide powder; dissolving the obtained enriched-selenium crude polysaccharide powder into 20-fold hot distilled water; adding trypsase and papain, taking the obtained protease treatment fluid, and adjusting the pH value to be 7.0 by using an oxalic acid to process; then taking supernate, adding an Sevag reagent to the obtained supernate, thereof obtaining deproteinated enriched-selenium polysaccharide; and preparing a 0.5% polysaccharide solution from the obtained deproteinated enriched-selenium polysaccharide, so as to obtain enriched-selenium polysaccharide quality goods.

Description

technical field [0001] The invention relates to a selenium-resistant bacterial strain domestication, selenium-enrichment research, a method for preparing selenium-enriched polysaccharide by using the bacteria, and the application of the selenium-enriched polysaccharide in improving immune regulation function and anti-oxidation function, which belongs to the technical field of microbiology and bioengineering . Background technique [0002] As an important trace element, selenium has been paid more and more attention by the scientific community. Studies have shown that selenium has the functions of improving the body's immunity, anti-oxidation, detoxification, inhibiting tumors, and preventing cardiovascular diseases. [0003] The toxicity of inorganic selenium compounds is much higher than that of organic selenium compounds, and their bioavailability is low. Only by converting inorganic selenium into organic selenium can it have edible and health care value. [0004] The b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/04C08B37/00A23L1/29A23K1/16C12R1/01A23K20/163A23L33/125
Inventor 汪以真徐春兰靳明亮姚国佳
Owner ZHEJIANG UNIV
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