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A kind of preservation method of phospholipase c

A technology of phospholipase and phosphate buffer, applied in the biological field, can solve the problems of phospholipase C activity loss, high energy consumption, etc., and achieve the effects of avoiding the loss of activity, reducing production energy consumption, and improving the stability of enzyme activity

Active Publication Date: 2020-09-18
WILMAR SHANGHAI BIOTECH RES & DEV CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Phospholipase C preparations are generally stored at 4°C. However, in the practical application of enzymatic degumming, phospholipase C is often placed in a separate storage tank for standby. Due to the relatively high ambient temperature, large energy consumption, and the loss of activity of phospholipase C is difficult to avoid if it is not refrigerated

Method used

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  • A kind of preservation method of phospholipase c
  • A kind of preservation method of phospholipase c
  • A kind of preservation method of phospholipase c

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1, the preparation of phospholipase C

[0037] 1. Construction of engineering bacteria

[0038] The starting strain is Pichia pastoris SMD1168 (purchased from Invitrogen Company), according to the Pichia expression kit (Invitrogen), the phospholipase C gene (GENBANK WP_000731010) of Bacillus cereus (Bacillus cereus) was transferred to, and the construction project bacteria as production strains.

[0039] 2. Fermentation of Phospholipase C

[0040] The medium and solution used therein consist of the following:

[0041] Seed shake flask medium: yeast extract (10g / L), tryptone (20g / L), glycerol (20g / L).

[0042] Fermentation basal medium: glycerol (50g / L), calcium sulfate (0.9g / L), potassium sulfate (14.67g / L), magnesium sulfate (11.67g / L), ammonium sulfate (9g / L).

[0043] Trace element PTM1 solution: CuSO4·5H2O(6.0g / L), NaI(0.08g / L), MnSO4·H2O(3.0g / L), Na2Mo4·2H2O(0.2g / L), ZnCl2·5H2O(20.04g / L) L), FeSO4 7H2O (65.05g / L), H3BO3 (0.02g / L), H2SO4 (19.2ml / L), ...

Embodiment 2、40

[0052] Example 2, Preservation of Phospholipase C at 40°C

[0053] The phospholipase C preparation obtained in Example 1 was stored at 40° C., and samples were taken out every 5 days to detect its remaining enzyme activity. The test results are shown in Table 1.

[0054] Table 1

[0055]

[0056]

[0057] The results showed that when phospholipase C was stored at 40°C, the enzyme activity of phospholipase C remained basically unchanged within 40 days of storage, indicating that phospholipase C had good storage stability when stored at 40°C.

Embodiment 3、38

[0058] Example 3, Preservation of Phospholipase C at 38°C

[0059] The phospholipase C preparation obtained in Example 1 was stored at 38° C., and samples were taken out every 5 days to detect its remaining enzyme activity. The test results are shown in Table 2.

[0060] Table 2

[0061]

[0062] The results showed that when phospholipase C was stored at 38°C, the enzyme activity of phospholipase C remained basically unchanged within 30 days of storage, indicating that phospholipase C had good storage stability when stored at 38°C.

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Abstract

The invention provides a preservation method of phospholipase C. By using the preservation method, the phospholipase C has good enzyme activity stability. When the preservation method of the phospholipase C provided by the invention is used in the practical application process of the phospholipase C, the production energy consumption can be reduced; and meanwhile, the activity loss of the phospholipase C is avoided.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for preserving phospholipase C. Background technique [0002] Phospholipases have the ability to hydrolyze one or more glycerophospholipid ester bonds and represent a class of lipases, acyl hydrolases and phosphatases. Phospholipase can be divided into phospholipase A1 (PLA1), phospholipase A2 (PLA2), phospholipase C (PLC) and phospholipase D (PLD) ( figure 1 ). [0003] Phospholipase C (Phospholipase C, referred to as PLC) is a lipolytic enzyme capable of hydrolyzing the phosphatidyl bond at the C3 site of glycerophospholipids to generate diglycerides, phosphorylcholine, phosphoinositol, and phosphoethanolamine. Phospholipase C widely exists in animals, plants and microorganisms. Animal and plant-derived PLC is generally located on the cell membrane with a relatively complex structure. It belongs to endogenous phospholipase C and is difficult to separate. The structure o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/16
Inventor 顾思天包悦佚关惠琴
Owner WILMAR SHANGHAI BIOTECH RES & DEV CENT
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