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Lipase stabilizing agent

A technology of lipase and stabilizer, which is applied in the field of enzyme preparations, can solve problems such as difference in effect and achieve a good stable effect

Inactive Publication Date: 2014-05-07
WILMAR SHANGHAI BIOTECH RES & DEV CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] However, for different enzymes, the composition and content of the stabilizer vary widely, and when the same stabilizer is used for different enzymes, its effect will also vary greatly

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0124] Embodiment 1, Pichia pastoris fermentation lipase preparation

[0125] 1.1, Pichia pastoris fermentation

[0126] Fermentation and cultivation of Pichia pastoris GS115 to prepare Pichia pastoris fermentation broth, the specific process is as follows:

[0127] The recombinant Pichia pastoris production strain WILMAR-RML was cultured in the seed shake flask medium at 30°C for 20h, and then inoculated into a solution containing sodium hexametaphosphate (25g / L) and trace elements PTM1 ( 4ml / L) in the basal medium. After a period of adaptation, the strain enters the exponential growth phase. Concentrated ammonia water is added automatically to control the pH5.5. When the glycerol in the fermentation medium is exhausted, DO rises sharply at this time, and 500g / L PTM1 containing 12ml / L is started to be fed. L of glycerol feeding solution, the temperature was controlled at 30°C.

[0128] When the wet weight of the cells reaches 220-250g / L, stop adding glycerin, and then add ...

Embodiment 2

[0139] Embodiment 2, stabilizer screening

[0140] Different stabilizers were added to the liquid Rhizomucor miehei lipase preparation obtained in Example 1, and the liquid Rhizomucor miehei lipase preparation that was added or not added to the stabilizer was stored at 25°C for 18 days. On the 3rd, 9th and 18th day, a small amount of liquid Rhizomucor miehei lipase preparation was diluted 4000 times, the enzyme activity was detected, and the preservation rate of the enzyme activity was calculated. The results are shown in Table 2.

[0141] The impact of table 2 different stabilizers on the thermal stability of liquid Rhizomucor miehei lipase

[0142]

[0143]

[0144] The experimental results show that: using Na 2 SO 4 , (NH 4 ) 2 SO 4 , sucrose, trehalose, Dextran-10, glycerol and sorbitol were used as stabilizers of Rhizomucor miehei lipase. After storage for 18 days at room temperature (25°C), the enzyme activity preservation rate reached more than 80%, while the...

Embodiment 3

[0145] Embodiment 3, composite stabilizer

[0146] 3.1. Screening of compound stabilizers

[0147] With the stabilizer Na screened out in embodiment 2 2 SO 4 , (NH 4 ) 2 SO 4 , sucrose, trehalose, Dextran-10, glycerin and sorbitol are compounded, and according to the method and concentration of Example 2, a suitable compound stabilizer is screened, wherein the storage temperature is 40 ° C, in order to carry out the accelerated test, the specific process is as follows :

[0148] The above-mentioned liquid Rhizomucor miehei lipase preparation added with a stabilizer was stored at 40°C for 18 days respectively, and on the 3rd day, 9th day and 18th day, a small amount of liquid Rhizomucor miehei lipase preparation was taken and diluted Enzyme activity was measured after 4000 times.

[0149] The results showed that at 40°C, when Rhizomucor miehei lipase was preserved using the compound solution of "sodium sulfate + sorbitol", the preservation rate of enzyme activity was muc...

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Abstract

The invention provides a lipase stabilizing agent. The lipase stabilizing agent comprises Na2SO4 and sorbitol. The lipase stabilizing agent has good effects of stabilization of rhizomucor mieheilipase and provides a basis for preservation and further application.

Description

technical field [0001] The invention relates to the field of enzyme preparations, in particular to a lipase stabilizer. Background technique [0002] Lipase is an enzyme that widely exists in animals, plants and microorganisms. Because microorganisms have the characteristics of simple cultivation, short cycle and large output, the enzyme preparations for industrial use are basically derived from microorganisms. [0003] Rhizomucor miehei (Rhizomucor miehei) lipase (RML) has high sn-1, 3 selectivity, and is widely used in the enzymatic synthesis of structured lipids, such as the synthesis of SOS, OPO, and DAG. As far as the current industrial application prospect is concerned, the unit yield of RML produced by natural Rhizomucor miehei is low, and the composition is unstable, the extraction and purification process is complicated, and the production cost is high, which limits its application. Therefore, using genetic engineering technology, heterologous expression of Rhizomu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/96
CPCC12N9/96C12N9/20C12Y301/01003
Inventor 顾思天关惠琴高霓思常桂芳
Owner WILMAR SHANGHAI BIOTECH RES & DEV CENT
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