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Strain for producing low-temperature sterol esterase and fermentation method thereof

A technology of sterol esterase and fermentation method, which is applied in the field of obtaining sterol esterase by fermentation of low-temperature strains, and achieves the effects of satisfying industrial production needs, wide application value and reducing production cost.

Inactive Publication Date: 2018-11-06
DALIAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no research reports on the production of low-temperature sterol esterase by marine microbial fermentation at home and abroad. If the industrial production of marine microbial fermentation sterol esterase can create good economic and social benefits

Method used

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  • Strain for producing low-temperature sterol esterase and fermentation method thereof
  • Strain for producing low-temperature sterol esterase and fermentation method thereof
  • Strain for producing low-temperature sterol esterase and fermentation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] The acquisition of low-temperature sterol esterase-producing bacteria Q-06:

[0039] 1) Original strain Pseudomonas fragi (CGMCC NO.1.3349) activation: activation medium: tryptone 8.0~10.0 g, yeast powder 4.0 g~9.0 g, NaCl 8.0~10.0 g, agar 18.0~20.0 g, tap water 1.0 L, pH value 6.0 ~8.0, sterilized by high pressure steam at 0.1MPa, 121℃ for 30 minutes.

[0040] 2) Low-temperature acclimation: low-temperature acclimatization of the strain in 1) to obtain low-temperature sterol esterase-producing strain Q-06, the acclimatization temperature from high to low is: 30°C → 28°C → 26°C → 24°C → 22°C → 20°C → 18°C to make it grow well in a low temperature environment; the low temperature acclimation medium used is: phytosterol ester 10.0~30.0g, yeast powder 5.0~10.0g, KH 2 PO 4 2.0~5.0g, MgSO 4 ·7H 2 O 0.5~1.5 g, (NH 4 ) 2 SO 4 1.0-3.0 g, glucose 5.0-15.0, agar powder 18-20.0 g, tap water 1.0 L, pH value 7.0, sterilized at 0.1 MPa, 121 °C for 30 min by high-pressure st...

Embodiment 2

[0043] Isolation and identification of low temperature sterol esterase producing bacteria Q-06:

[0044] The morphological characteristics of the colony are:

[0045] It grows well on the sterol ester medium. The single colony of the strain Q-06 is approximately round, with neat edges, raised surface, light yellow, opaque, easy to pick with an inoculation loop, and the center is shiny (see figure 1 );

[0046] The morphological characteristics of the bacteria are:

[0047] The strains are rod-shaped, mostly in a single arrangement, and Gram-stained results are negative (see figure 2 );

[0048] Main physiological and biochemical characteristics:

[0049] Oxidase test and contact enzyme test are positive, can oxidize and decompose glucose, can generate indole acetic acid, cannot generate hydrogen sulfide, cannot hydrolyze starch, nitrate reduction test, VP test, methyl red (MR) test, The gelatin hydrolysis test was negative, and the arginine dihydrolase test was positive...

Embodiment 3

[0056] The fermentation process of the low-temperature sterol esterase producing bacteria Q-06 strain:

[0057] 1) Culture medium preparation

[0058] ① Strain activation medium: tryptone 8.0g, yeast powder 5.0g, NaCl 8.0g, agar powder 20.0g, tap water 1.0L, pH value 6.5, sterilize at 0.1MPa, 121℃ for 30 minutes by high-pressure steam;

[0059] ② Liquid seed medium: yeast extract 8.0 g, peptone 3.5 g, NaCl 7.0 g, K 2 HPO 4 2.0g, FeSO 4 ·7H 2 O0.1g, MgSO 4 ·7H 2 O 0.2g, 1.0 L tap water, pH value 8.0, sterilize at 0.1 MPa, 121℃ for 30 min by high-pressure steam;

[0060] ③ Fermentation enzyme production medium: phytosterol ester 5.0g, yeast powder 3.0g, KH 2 PO 4 1.0g, MgSO 4 ·7H 2 O 0.3g, (NH 4 ) 2 SO 4 0.5g, glucose 5.0g, tap water 1.0L, pH value 6.0, sterilize at 0.1MPa, 121℃ for 30min by high-pressure steam.

[0061] 2) The strain Q-06 producing low-temperature sterol esterase was initially activated according to the strain instructions provided by China Gen...

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Abstract

The invention relates to the fields of microbiology, biochemistry and fermentation engineering, and discloses a strain for producing low-temperature sterol esterase, and the strain is named as Q-06, and is derived from low temperature acclimation of original strain Pseudomonas fragi. The optimum temperature for producing the low-temperature sterol esterase by use of the strain Q-06 is 18-24 DEG C,the low-temperature sterol esterase can be prepared without heating / cooling and the like to consume energy, production cost is reduced, industrial production needs can be met, the strain Q-06 for producing the low-temperature sterol esterase has the ability to produce a sterol esterase, and efficiently produces the stable low-temperature sterol esterase.

Description

technical field [0001] The invention relates to the fields of microbiology, biochemistry, fermentation engineering and the like, in particular to a method for acclimating a Pseudomonas berry strain at low temperature and obtaining sterol esterase by fermenting the new low-temperature bacterial strain. Background technique [0002] Sterol esterase (Sterol esterase, EC 3.1.1.13) is an ester hydrolase that can catalyze the hydrolysis of sterol esters to generate sterols and fatty acids. Under suitable conditions, it can also catalyze the synthesis of sterol esters through esterification or transesterification reactions. Sterol esterase can help the body digest and absorb oil and fat nutrients, because it is related to human lipid metabolism and cholesterol absorption, sterol esterase is one of the important enzyme preparations for detecting the total cholesterol content in blood. In the paper industry, the combination of sterol esterase and lipase can be used to effectively hyd...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/18C12R1/38
CPCC12N1/20C12N9/18C12Y301/01013C12N1/205C12R2001/38
Inventor 张庆芳任楠楠王晓辉迟乃玉
Owner DALIAN UNIV
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