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Low-temperature lipase mycopremna, low-temperature lipase and method of preparing the same

A production method, lipase technology, applied in the direction of fungi, hydrolytic enzymes, etc.

Inactive Publication Date: 2011-05-11
THE INST OF MICROBIOLOGY XINJIANG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the low-temperature lipase studied is mainly produced by cold-tolerant microbial strains, and there are few strains available for research

Method used

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  • Low-temperature lipase mycopremna, low-temperature lipase and method of preparing the same
  • Low-temperature lipase mycopremna, low-temperature lipase and method of preparing the same
  • Low-temperature lipase mycopremna, low-temperature lipase and method of preparing the same

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Experimental program
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Effect test

Embodiment 1

[0074] Embodiment 1: the cultivation-I of low-temperature lipase producing bacteria (GL-1)

[0075] The low-temperature lipase producing bacteria GL-1 of the present invention is inoculated in the fat assimilation plate medium, wherein (NH 4 ) 2 SO 4 0.1%;K 2 HPO 4 0.1%; NaCl 0.05%; MgSO 4 ·7H 2 O 0.05%; FeSO 4 ·7H 2 O 0.001%; ​​yeast extract 0.5%; agar 2.5%; natural pH; take olive oil and 2% polyvinyl alcohol (PVA) to mix in a ratio of 1:3, 10000r / min stirring and emulsifying for 5min, 0.8kg / cm 2 After sterilizing for 30 minutes, take 12ml and add it to 100ml of the above medium. After culturing at about 20°C for 5-7 days, a transparent circle can be formed around the colony of the low-temperature lipase-producing bacteria of the present invention.

Embodiment 2

[0076] Embodiment 2: Cultivation-II of low-temperature lipase-producing bacteria (GL-1)

[0077] Rhodamine B was prepared as a 0.05% mother solution, and added to the oil assimilation medium of Example 1 in an amount of 1.0%, to prepare a Rhodamine B chromogenic medium. When the low-temperature lipase-producing bacteria GL-1 of the present invention is inoculated in the above-mentioned culture medium, an orange-red halo will appear around the colonies of the low-temperature lipase-producing bacteria of the present invention, and red fluorescent circles will appear under ultraviolet light.

Embodiment 3

[0078] Embodiment 3: the fermentation culture of low-temperature lipase producing bacteria (GL-1 bacterial strain)

[0079] Insert the low-temperature lipase-producing bacteria of the present invention into the wort liquid seed medium, wherein malt extract powder 0.3; yeast extract 0.3; peptone 0.5; glucose 1.0; 1.0kg / cm 2 Steam sterilize for 15 minutes. After culturing at 20°C for 1-2 days, inoculate in the following fermentation medium with a 5% inoculation amount. Bean cake powder 2.0, corn syrup 2.0, glucose 1.0, K 2 HPO 4 0.5, NaNO 3 0.5, pH7.0, in 250ml Erlenmeyer flask, 50ml per bottle, 0.8kg / cm 2 Steam sterilize for 30 minutes. Shake culture at 20°C and 200rpm for 48h. After culturing, the bacterium is removed by centrifugation to obtain a low-temperature lipase crude enzyme liquid, and the fermented enzyme yield can reach 20 U / mL. Add ammonium sulfate to the lipase solution obtained in this way to make its saturation reach 60%, let it stand for precipitation to...

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Abstract

The invention discloses a Trichosporon pullulans which generates a low-temperature lipase and a method for manufacturing the low-temperature lipase through the Trichosporon pullulans, as well as provides the low-temperature lipase, which is characterized by: utilizing the Trichosporon pullulans (CGMCC. No. 1674) to build a bacterial preservation, rejuvenation and breeding system method; determining a stable ferment method through optimizing and sieving a culture medium formula and optimizing and controlling the fermenting technique. The manufactured low-temperature lipase has good low-temperature activity and high-enzyme activity, widely suits for washing, food processing, leather processing, digestant, organic synthetic chemical product, perfume, drug preparation, fat processing industries and so on, and has important significance.

Description

technical field [0001] The invention relates to a low-temperature lipase, a bacteria producing the enzyme and a production method thereof. Specifically, the present invention relates to a lipase produced by low-temperature-resistant microbial strains and having higher activity at lower temperatures, and a production method for producing low-temperature lipase using the microbial strains. technical background [0002] Lipase is an enzyme ubiquitous in nature. According to its different sources, it can be divided into three categories: animal lipase, plant lipase and microbial lipase. Among them, microbial lipase is widely used in washing, food processing, leather processing, digestive aids, organic synthetic chemical products, spices, and pharmaceutical preparations because of its easy production, easy expansion, and its specific decomposition characteristics for fatty substances. , oil processing and other aspects have been widely used, becoming one of the world's main in...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/16C12N9/20
Inventor 谢玉清顾美英唐琦勇茆军张志东冯怀蓉
Owner THE INST OF MICROBIOLOGY XINJIANG ACADEMY OF AGRI SCI
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