Preparation method and application of thermomyces lanuginosus lipase

A technology of Thermomyces lanuginosus and lipase, which is applied in the field of the preparation of Thermomyces lanuginosus lipase, lipase, and Thermomyces lanuginosus lipase, which can solve the problem of high culture temperature

Active Publication Date: 2013-08-14
SHANDONG YOUTELL BIOCHEM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0025] Thermomyces lanuginosus is a wide-spread fungus with a high growth upper limit temperature, which can produce a thermostable and industrially important lipase, Thermomyces lanuginosus lipase, but wild There are many unfavorable factors such as high culture temperature for bacterial enzyme production. Therefore, in recent years, the main research direction has focused on the use of genetic engineering methods to construct recombinant bacteria, and then express lipase
At present, there is no relevant report about using Trichoderma reesei to express Thermomyces lanuginosus lipase

Method used

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  • Preparation method and application of thermomyces lanuginosus lipase
  • Preparation method and application of thermomyces lanuginosus lipase
  • Preparation method and application of thermomyces lanuginosus lipase

Examples

Experimental program
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Effect test

Embodiment 1

[0048] The preparation of embodiment 1 thermomyces lanuginosus lipase

[0049] 1. Microbial and plasmid materials:

[0050] Escherichia coli DH5α used for cloning was provided by Invitrogen. The host used to provide lipase gene resources and high-efficiency expression was Trichoderma reesei RUTC-30, purchased from AmericanTypeCultureCollection (ATCC), and the starting plasmid pBluescriptKS (1) was purchased from Stratagene.

[0051] 2. Artificial synthesis of Thermomyces lanuginosus lipase (TTL) gene and construction of expression plasmid:

[0052] Commissioned Life Technologies Corporation to artificially synthesize the lipase (LipA) gene (SEQ ID NO.1) encoding Thermomyces lanuginosa lipase (LipA) using conventional chemical methods in the field. The synthetic gene code is more in line with the highly expressed gene code of Trichoderma reesei. For the convenience of cloning, a SpeI restriction site was added at the 5' end and an XhoI restriction site was added at the 3' end...

Embodiment 2

[0061] Example 2 Improvement experiment of steamed bread whiteness

[0062] Add 0.4g of yeast and 23ml of water to the dough mixing bowl. After activation, add 50g of flour and lipase for experiment (addition amount is as shown in Table 2), and knead the dough by hand (make sure the kneading time, kneading frequency and intensity are consistent) , After the dough is formed, press it on the noodle machine for 12 times, let it stand for 2 minutes, make a ball by hand, proof it in the proofing box for 35 minutes, steam it in the steamer for 18 minutes, and check it after cooling.

[0063] The quality evaluation of steamed buns mainly refers to the "Manufacturing and scoring standard of wheat flour for steamed buns (steamed buns)" in SB / T10139-93. The main evaluation indicators include the surface whiteness, volume and internal structure of steamed buns. The specific evaluation methods are as follows:

[0064] Surface whiteness: use a whiteness meter to test, take 4 different points...

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Abstract

The invention discloses a preparation method of a thermomyces lanuginosus lipase. The method comprises the following steps of: 1) synthesizing a thermomyces lanuginosus lipase gene; 2) constructing a thermomyces lanuginosus lipase expression plasmid; 3) converting a trichodermareesei and selecting high-yield strains of the thermomyces lanuginosus lipase; and 4) separating and purifying the thermomyces lanuginosus lipase. The invention also discloses the application of the thermomyces lanuginosus lipase to the preparation of flour foods.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a lipase, in particular to a lipase from Thermomyces lanuginosus; in addition, the present invention also relates to a preparation method and a method for the Thermomyces lanuginosus lipase. application. Background technique [0002] Lipase widely exists in animals, plants and microorganisms. Plants that contain more lipase are the seeds of oil crops, such as castor bean and rapeseed. When the oil seeds germinate, lipase can work together with other enzymes to catalyze and decompose oily substances, providing the seeds necessary for rooting and germination. Nutrients and energy; more lipase in animals is the pancreas and adipose tissue of higher animals, a small amount of lipase is contained in the intestinal juice, which is used to supplement the lack of fat digestion by pancreatic lipase, and the gastric juice of carnivorous animals contains A small amount of butyrin este...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/20C12N15/80A21D8/04C12R1/885
Inventor 李新良张珊嘉张民唐鹏段园园马天画王锡珍
Owner SHANDONG YOUTELL BIOCHEM
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