A strain of Pseudomonas with high trehalose synthase production and its fermentative enzyme production method

A technology of trehalose synthase and Pseudomonas, which is applied in the field of microorganisms or enzymes to achieve the effects of reducing fermentation costs, increasing fermentation production capacity, and improving production efficiency

Active Publication Date: 2020-06-23
SHANDONG LONGKETE ENZYME PREPARATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, due to the restriction of enzyme activity and enzyme application conditions, the prior art trehalose synthase still cannot meet the needs of industrial production, so a high-yield trehalose synthase Pseudomonas strain is selected, and its fermentation product has good Excellent heat resistance and low production cost, it is of great significance to promote the mechanization of various industries on a large scale

Method used

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  • A strain of Pseudomonas with high trehalose synthase production and its fermentative enzyme production method
  • A strain of Pseudomonas with high trehalose synthase production and its fermentative enzyme production method
  • A strain of Pseudomonas with high trehalose synthase production and its fermentative enzyme production method

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Effect test

Embodiment 1

[0036] Example 1 Pseudomonas CSY-67 Fermentation Enzyme Production and Extraction and Purification Method of Trehalose Synthase Produced

[0037] The optimum action pH of trehalose synthase produced by the fermentation of Pseudomonas CSY-67 is 7.5, and the optimum action temperature range is 35°C.

[0038] The fermentative enzyme production method of Pseudomonas CSY-67 mainly comprises the following steps:

[0039] Slant culture: Pick a ring of Pseudomonas CSY-67 and inoculate it on a solid slant medium, and culture at a constant temperature of 37°C for 48 hours to obtain first-grade seeds;

[0040] Shake flask culture: Take a ring of the first-class seeds and insert them into the seed medium, and cultivate them for 48 hours at a constant temperature of 37°C and a shaker speed of 200 r / min to obtain the second-class seed liquid;

[0041] Seed tank culture: put the secondary seed liquid into the seed tank medium at a ratio of 5% (v / v) of the inoculation amount, and cultivate f...

Embodiment 2

[0055] Example 2 Pseudomonas CSY-67 Fermentation Enzyme Production and Extraction and Purification Method of Trehalose Synthase Produced

[0056] The optimum action pH range of trehalose synthase produced by Pseudomonas CSY-67 fermentation is 8.5, and the optimum action temperature range is 45°C.

[0057] The fermentative enzyme production method of Pseudomonas CSY-67 mainly comprises the following steps:

[0058] Slant culture: Pick a ring of Pseudomonas CSY-67 and inoculate it on a solid slant medium, and culture at a constant temperature of 37°C for 48 hours to obtain first-grade seeds;

[0059] Shake flask culture: Take a ring of the first-class seeds and insert them into the seed medium, and cultivate them for 48 hours at a constant temperature of 37°C and a shaker speed of 200 r / min to obtain the second-class seed liquid;

[0060] Seed tank culture: put the secondary seed solution into the seed tank culture medium at a ratio of 5% (v / v) of the inoculum, and cultivate fo...

Embodiment 3

[0074] Example 3 Pseudomonas CSY-67 Fermentation Enzyme Production and Extraction and Purification Method of Trehalose Synthase Produced

[0075] The optimum action pH range of trehalose synthase produced by Pseudomonas CSY-67 fermentation is 8, and the optimum action temperature range is 40°C.

[0076] The fermentative enzyme production method of Pseudomonas CSY-67 mainly comprises the following steps:

[0077] Slant culture: Pick a ring of Pseudomonas CSY-67 and inoculate it on a solid slant medium, and culture at a constant temperature of 37°C for 48 hours to obtain first-grade seeds;

[0078] Shake flask culture: Take a ring of the first-class seeds and insert them into the seed medium, and cultivate them for 48 hours at a constant temperature of 37°C and a shaker speed of 200 r / min to obtain the second-class seed liquid;

[0079] Seed tank culture: the secondary seed solution was added to the seed tank culture medium at a ratio of 5% (v / v) of the inoculum, and cultivated...

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Abstract

The invention discloses a strain of Pseudomonas with high trehalose synthase production and its fermentative enzyme production method, which belongs to the field of microorganisms or enzymes. The strain is specifically Pseudomonas sp. CSY‑67, with a preservation number It is CGMCC No.13154. The strain was submerged in a liquid 50L tank, fed with feed, and fermented for 80 hours. The average level of enzyme production was 39575U / mL. According to the method for liquid-state fermentation of trehalose synthase established by the present invention, the optimum pH range of the produced trehalose synthase is 7.5-8.5, the optimum action temperature range is 35°C-45°C, and the remaining enzyme activity after incubation at 60°C for 1 hour is 88%, the remaining enzyme activity is 59.5% after being incubated at 80°C for 2 hours, has good heat resistance, and can be widely used in the industrial production of trehalose.

Description

technical field [0001] The invention belongs to the field of microorganisms or enzymes, and in particular relates to a strain of Pseudomonas with high trehalose synthase production and a fermentative enzyme production method thereof. Background technique [0002] Trehalose is a non-reducing disaccharide formed by connecting two molecules of glucose with 1,1-glycosidic bonds. It is widely found in organisms such as bacteria, yeast, filamentous fungi, plants, insects, and invertebrates. Trehalose has very important biological significance to organisms. It is a reserve of energy and carbon sources. It is a stabilizer and protector for proteins and biofilm molecules in harsh environments such as dehydration, high temperature, oxygen free radicals, and low temperature. It is a signal-sensing complex and a growth regulator, and it is also a component of some bacterial cell walls. Because trehalose has special properties such as moderate sweetness, stable properties, not easy to d...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/90C12R1/38
CPCC12N9/90C12Y504/99016C12N1/205C12R2001/38
Inventor 王兴吉刘文龙曹世源张杰
Owner SHANDONG LONGKETE ENZYME PREPARATION
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