A Bacillus subtilis Mutation Strain Producing Phospholipase

A technology of Bacillus subtilis and phospholipase is applied in the field of enzymatic degumming, which can solve problems such as oil degumming and achieve good degumming effect.

Active Publication Date: 2019-08-13
WILMAR SHANGHAI BIOTECH RES & DEV CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the existing phospholipase C producing strains are pathogenic strains, so it is impossible to directly use the existing phospholipase C producing strains to degumming oil

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Embodiment 1, mutagenesis selection of bacterial strain

[0073] Inoculate Bacillus subtilis WBRD01280 (Accession No.: CGMCC No.7506) into a shaker flask containing 50ml of LB liquid medium, culture at 180rpm, 28°C for 16h, and reach the logarithmic growth phase to obtain Bacillus subtilis WBRD01280 bacterial liquid.

[0074] Take 1ml of Bacillus subtilis WBRD01280 bacteria liquid, centrifuge at 12000rpm for 2 minutes, discard the supernatant, dilute and dissolve with 1ml of sterile water and wash the centrifuged sediment twice, centrifuge at 12000rpm for 2 minutes, collect the cleaned bacteria sludge, and then use sterile water Dilute the bacteria sludge so that the OD600nm value of the bacteria suspension is between 0.5-0.9. Take 1ml of the bacterial suspension and add it to the Erlenmeyer shaker flask with glass beads, then add 30μL of 50wt% diethyl sulfate solution, shake at 28°C for 17min, then immediately add 0.4ml of 25wt% sodium thiosulfate solution. Take 1ml o...

Embodiment 2

[0080] Example 2, Preparation of Bacillus subtilis Mutagen WBRD01281-D Extracellular Phospholipase C

[0081] The Bacillus subtilis mutant strain WBRD01281-D and Bacillus subtilis WBRD01280 were respectively inoculated into LB medium, and cultured at 28° C. and 180 rpm for 24 hours to obtain WBRD01281-D seed liquid and Bacillus subtilis WBRD01280 seed liquid.

[0082] The WBRD01281-D seed liquid and the Bacillus subtilis WBRD01280 seed liquid were inoculated into the fermentation medium at an inoculum amount of 2%, and cultured at 28°C and 180 rpm for 24 hours to obtain the WBRD01281-D fermentation liquid and the Bacillus subtilis WBRD01280 fermentation liquid.

[0083] The fermentation broth was centrifuged at 12000 rpm for 2 minutes to obtain the centrifuged supernatant of WBRD01281-D and WBRD01280 respectively.

[0084] The phospholipase C enzyme activity of WBRD01281-D centrifugal supernatant and Bacillus subtilis WBRD01280 centrifugal supernatant were detected respectivel...

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PUM

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Abstract

A phospholipase-producing mutagenic strain of Bacillus subtilis. The invention provides an isolated bacterial strain expressing phospholipase C, which is preserved in the General Microorganism Center of China Microbiological Culture Collection Management Committee, and the preservation number is CGMCC No.8500. The phospholipase C prepared by using the bacterial strain and the preparation method provided by the invention can be used for oil refining, has good degumming effect, and can be widely used in the fields of oil refining, additives, medicine and the like.

Description

technical field [0001] The invention belongs to the field of enzymatic degumming, in particular to a Bacillus subtilis mutagenic strain producing phospholipase. Background technique [0002] Phospholipase (Phospholipase, PL) is an enzyme that can hydrolyze glycerophospholipids in organisms. The hydrolyzed products are various phosphatidic acids and amino alcohols, such as cholamine, choline, serine, ethanolamine, etc. Phospholipase can be divided into phospholipase A (phospholipase A, PLA), phospholipase B (Phospholipase B, PLB), phospholipase C (Phospholipase C, PLC) and phospholipase D (Phospholipase D, PLD). Phospholipase C acts on the 3-position phosphate bond, and the product is diglyceride, phosphorylcholine or phosphoethanolamine, etc. [0003] Phospholipase can be widely used in oil refining, phospholipid modification, feed improver, food industry and medicine. Degumming is an important link in the refining process of vegetable oil, which is crucial to improving t...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/16C11B3/00C12R1/125
CPCC11B3/003C12N9/16C12Y301/04003C12N1/205C12R2001/125
Inventor 周美凤徐正军许骏
Owner WILMAR SHANGHAI BIOTECH RES & DEV CENT
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