Nie's brevibacterium new strain and process for preparing enzyme by it

The technology of Brevibacterium nesoi and new strains is applied in the field of preparation of new strains of Brevibacterium nesoi and enzymes produced by them, which can solve the problems of complex extraction process, difficulty in large-scale production and high cost, and achieves high activity, The cultivation method is simple, easy to implement, and the effect of low cost

Inactive Publication Date: 2006-06-28
WUHAN CHEM COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although alcohol dehydrogenase and acetaldehyde dehydrogenase can be extracted from the liver, pancreas and other viscera of some animals, the extraction process is relatively complicated, the cost is high, and it is not easy to produce on a large scale

Method used

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  • Nie's brevibacterium new strain and process for preparing enzyme by it
  • Nie's brevibacterium new strain and process for preparing enzyme by it
  • Nie's brevibacterium new strain and process for preparing enzyme by it

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Weigh 5g NaCl, 10g peptone, 3g beef extract, 1000ml distilled water, 16g-20g agar to prepare beef extract medium;

[0020] Prepare 1% (0.01g soil / ml water by weight) soil solution, gradient dilution, preparation concentration is 10 -5 g / ml soil solution, take 200μl diluted soil solution and add it to 30ml beef extract medium, add 200μl~400μl ethanol inducer at the same time, and then cultivate this culture at 35℃~37℃ for 16h~36h; 4 ~5 times of isolation and purification culture, microscopic examination as a single strain, and preservation at 4°C on a slant. The physiological and biochemical characteristics of the strain are shown in Table 1.

[0021] Connect the strains obtained by screening to 30ml (250ml Erlenmeyer flask) fermented liquid containing 5gNaCl, 10g peptone, 3g beef extract, and 1000ml distilled water from the inclined plane, add 200μl~400μl ethanol inducer, at 35℃~37℃, 150r / Cultivate for 4d-6d (days) under min to obtain the acetaldehyde dehydrogenase w...

Embodiment 2

[0023] Weigh: 5g NaCl, 10g peptone, 3g beef extract, 1000ml distilled water, 16g-20g agar to prepare beef extract medium fermentation broth.

[0024] The preparation concentration is 10 -4 g / ml soil solution, take 200μl and add it to 30ml beef extract medium, and add 200μl~400μl ethanol inducer at the same time, and then cultivate the culture at 30℃~37℃ for 16h~36h. Carry out liquid fermentation according to the method of embodiment 1, then this fermented liquid ultrasonic breaks up, measures its activity after centrifuging and purifying. The acetaldehyde dehydrogenase activity of the strain was measured to be 8.3333U / ml after microscopic examination as a single strain.

Embodiment 3

[0026] Weigh: 5g NaCl, 10g peptone, 3g beef extract, 1000ml distilled water, 16g-20g agar to prepare beef extract medium fermentation broth.

[0027] The preparation concentration is 10 -4g / ml soil solution, take 200μl and add it to 30ml beef extract medium, and add 200μl~400μl ethanol inducer at the same time, and then cultivate the culture at 30℃~37℃ for 16h~36h. Carry out liquid fermentation according to the method of embodiment 1, then this fermented liquid ultrasonic breaks up, measures its activity after centrifuging and purifying. The acetaldehyde dehydrogenase activity of the strain was measured to be 6.5789U / ml after microscopic examination as a single strain.

[0028] Test items

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Abstract

The invention relates to brachybacterium nesterenkovi new strain and its culturing condition, physiological biochemical characteristic, and enzyme manufacturing method. It includes the following steps: preparing soil solution; gradient diluting; coating the given 200ul solution to 30ul beef extract culture medium; adding 100ul-800ul alcohol inducer [12-14]; culturing at 30-40 centigrade degree for 16-48h; purifying for many times; it is single cloning strain by microscopic examination; liquid fermenting; ultrasonic wave smashing; centrifugal separating; purifying; and testing its activity. The advantages of the invention are that the strain can produce acetadehyde dehydrogenase and alcohol dehydrogenase; its culturing condition is simple and low cost to be good for industrialization production; it has high activity at 37 centigrade degree and can impel alcohol to metabolize in human body.

Description

technical field [0001] The invention relates to a new strain of Brachybacterium nesterenkovii and its culture conditions, physiological and biochemical characteristics and a preparation method of the produced enzyme. The new bacterial strain was deposited in China Center for Type Culture Collection on July 20, 2005, with the preservation number: CCTCC NO: M 205113. Background technique [0002] The genus Brachybacterium was first discovered in 1988 (Collins M.D, Brown J, Jones D, et al. Brachybacterium faecium gene. Nov., sp. nov., a coryneform bacterium from poultry deep litter. Int. J .Syst.Bacteriol., 1988, 38:45-48.), there is no report in China at present. Among the genus Brachybacterium, 11 species have been discovered and have been assigned species, and 26 species have not been assigned species. The 11 species are B.alimentarium, B.conglomeratum, B.faecium, B.fresconis, B.nesterenkovii, B.paraconglomeratum, B.rhamnosum, B.sacelli, B.Tyrofermentans, Brachybacterium a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/04C12R1/13
Inventor 吴元欣薛永萍赵玉凤王存文池汝安朱圣东
Owner WUHAN CHEM COLLEGE
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