Process for extracting two kinds of yeast bilesalt hydrolase and production technology of functional posset

A technology of bile salt hydrolase and extraction method, applied in the production field of functional milk wine and kefir

Active Publication Date: 2008-12-31
BEIJING UNIV OF AGRI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

On September 11, 2006, it was preserved in the General Microorganism Center

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  • Process for extracting two kinds of yeast bilesalt hydrolase and production technology of functional posset
  • Process for extracting two kinds of yeast bilesalt hydrolase and production technology of functional posset
  • Process for extracting two kinds of yeast bilesalt hydrolase and production technology of functional posset

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Embodiment 1

[0029]Example 1. Screening and Identification of Kluyveromyces marxianus K1 and M3 Strains with High Efficiency in Lowering Cholesterol and High Production of Bile Salt Hydrolase

[0030] 1. Screening of high-efficiency cholesterol-lowering and high-production bile salt hydrolase strains

[0031] Take 5mL of the filtrate of Zangling mushroom and pour it into 45mL sterile saline with glass beads, shake it fully for 30min to make a bacterial suspension, and then dilute it 10 times to 10 -4 ~10 -6 diluted bacterial solution. The plates were inoculated by the dilution pouring method, poured into the potato lactose selective medium containing antibiotics, and cultured at 28°C for 2-3 days. According to the colony characteristics of the yeast and the results of microscopic examination of the water immersion slides, the target colonies on the above plate were streaked and inoculated on the corresponding slant medium, and cultured at 28°C for 2-3 days. Transfer to inoculate in pota...

Embodiment 2

[0051] Example 2, Utilizing Kluyveromyces marxe K1 and M3 to Extract Bile Salt Hydrolase

[0052] 1. Fermentation process conditions for high production of bile salt hydrolase

[0053] Medium components (w / v) for cultivating Kluyveromyces marxii K1 and M3 high-yielding bile salt hydrolase: lactose 2%, potato 20% (potatoes are peeled and cut into pieces, boiled with water for 10-20min, covered with double gauze) filter), agar 1.5%, distilled water 1000mL, 0.07MPa sterilization for 20min.

[0054] Fermentation process conditions for high production of bile salt hydrolase:

[0055] M3 strain: the fermentation temperature is 32° C., the fermentation time is 24 hours, the initial pH value of the medium is 6.0, and the inoculation amount is 1%.

[0056] K1 strain: the fermentation temperature is 32° C., the fermentation time is 18 hours, the initial pH value of the medium is 6.0, and the inoculation amount is 2%.

[0057] Under the optimized fermentation conditions, the enzyme ac...

Embodiment 3

[0067]The activity of the bile salt hydrolyzing enzyme is measured by the amount of amino acids released after the degradation of the combined bile salt. That is, the enzyme activity is directly proportional to the amount of amino acid produced. The content of amino acids can be determined by ninhydrin chromogenic method. Take 2mL of enzyme reaction solution, add 1mL of ninhydrin color reagent, mix well, heat in boiling water bath for 16min, cool with tap water, add 5mL of diluent, shake well, and use 2mL of distilled water instead of enzyme reaction solution as the blank control to adjust to 0 , Measure the absorbance value A at 570nm (the generated color is stable within 60min, preferably completed within 30min). The amino acid produced by bile salt hydrolase at 1 μmol per minute was defined as an enzyme activity unit. Example 3. Production of functional milk wine by utilizing bacterial strains that are highly effective in lowering cholesterol and producing bile salt hydro...

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Abstract

The invention relates to a method for extracting bile salt hydrolase (BSH) of two kinds of yeasts and a production technology of functional kefir, applicable to the production of functional kefir and sour kefir in fermented dairy products. The invention adopts high-throughput screening technique and phthalic aldehyde method to acquire two kinds of yeasts which can lower cholesterol efficiently and have higher capacity of fermenting lactose, and Oxford cup method to acquire strains yielding the BSH highly through secondary screening. The invention optimizes the fermentation conditions and extraction method for the two kinds of yeasts to produce the BSH, and adopts proper cracking method by ultrasonic wave, which solves the problem that the cell walls of the yeasts are difficult to be cracked, and the cracking effect can be compared with that of the high pressure cell cracker. Kluyveromyces Marxianus K1 (CGMCC No 1812) and M3 (CGMCC No 1811) which can lower the cholesterol efficiently can yield the BSH highly and can ferment lactose to yield ethanol highly, are adopted to prepare the kefir of high quality that has the function of lowering the cholesterol. The fermentation process of preparing the functional kefir is simple with shorter fermentation period and lower cost. Thus, the fermentation process is applicable to mass industrial production.

Description

technical field [0001] The invention relates to an extraction method of two kinds of yeast bile salt hydrolase (Bile Salt Hydrolase, BSH) and a production technology of functional milk wine, which is suitable for the production of functional milk wine and kefir wine in fermented dairy products. Background technique [0002] Probiotics refer to a class of microorganisms that improve the microecological balance of specific parts of the host through intestinal colonization and have several other beneficial physiological functions. They can improve the ecological balance of intestinal microorganisms, control intestinal infection, reduce serum cholesterol levels, improve the utilization of lactose for people with lactose intolerance, and can stimulate specific or non-specific immune responses, and have the effect of immune enhancement. The live bacteria of probiotic products should have stable metabolism and strong activity, survive in large numbers after passing through the dige...

Claims

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Application Information

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IPC IPC(8): C12N9/00C12N1/16
Inventor 刘慧
Owner BEIJING UNIV OF AGRI
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