A kind of lipase and its production method and application

A technology of lipase and seeds, applied in the field of microorganisms, to achieve the effect of improving application value, significant low temperature resistance and alkali resistance, and improving production efficiency

Active Publication Date: 2020-09-25
SHANDONG LONGKETE ENZYME PREPARATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the optimal reaction temperature of lipase in the prior art is generally 40°C or higher, and the inactivation is serious below 30°C. Therefore, subject to the restriction of enzyme activity and applicable conditions of the enzyme, the lipase in the prior art still cannot meet the requirements. Therefore, screening a lipase with good alkali resistance and low temperature resistance and low production cost is of great significance to promote the mechanization of various industries on a large scale

Method used

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  • A kind of lipase and its production method and application
  • A kind of lipase and its production method and application
  • A kind of lipase and its production method and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The mutagenesis selection of embodiment 1 Pseudomonas LD-14

[0041] Take the fresh slant of the two starting strains LA-32, wash the bacteria with sterile water, oscillate in a test tube with glass beads to disperse the bacteria, collect the bacteria by centrifugation, resuspend the bacteria with 5% glycerol, and count with blood cells Count the plates until the bacterial concentration is 10 7 -10 8 cells / mL, as the starting bacterial suspension.

[0042] Turn on the normal temperature and pressure plasma system, wipe the inside and outside of the operating room with alcohol cotton, and turn on the ultraviolet lamp for 30 minutes to sterilize. After the sterilization in the operating room of the system is completed, take 10 μL of the bacterial suspension and spot it on the rough surface of the slide, and transfer the slide to the table of the operating room with tweezers under aseptic conditions. Open the helium gas valve, set the gas flow and mutagenesis time for m...

Embodiment 2

[0054] Embodiment 2 Pseudomonas LD-14 fermentation produces enzyme

[0055] The fermentative enzyme production method of Pseudomonas LD-14 mainly comprises the following steps:

[0056] Slant culture: pick a ring of Pseudomonas LD-14 and inoculate it on a solid slant medium, and culture at a constant temperature of 35°C for 36 hours to obtain first-grade seeds;

[0057] Shake flask culture: take a ring of the first-grade seeds and insert them into the seed medium, and cultivate them for 48 hours at a constant temperature of 35°C and a shaker speed of 200r / min to obtain the second-grade seed liquid;

[0058] Seed tank culture: put the secondary seed solution into the seed tank culture medium at a ratio of 6% (v / v) of the inoculum, and cultivate for 12 hours at a constant temperature of 35°C and a rotational speed of 200r / min;

[0059] Fermentation tank culture: The seed liquid in the seed tank is inserted into the culture medium of the fermenter according to the ratio of the i...

Embodiment 3

[0070] Embodiment 3 Pseudomonas LD-14 fermentation performance verification

[0071] Carry out 50L fermenter verification experiment according to the fermentative enzyme production method of Pseudomonas LD-14 of embodiment 2, fermentation cycle 150h, its 6 batches of fermentative enzyme production situation, average enzyme production level is 25558U / mL, table 2 illustrates bacterial strain Not only high-yield lipase, but also the fermentation performance and the enzymatic activity of the lipase produced by it have remarkable stability.

[0072] Table 2 Fermentation and enzyme production of 6 batches of high lipase-producing strains

[0073]

[0074]

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Abstract

The invention belongs to the technical field of microbes, and in particular relates to lipase as well as a production method and application thereof. The lipase is produced from Pseudomonas sp. LD-14, and a collection number of the strain is CGMCC No. 14414. The enzyme activity of fermentation liquor for fermenting the Pseudomonas LD-14 to produce the lipase reaches 25000-26000U / mL; the optimal pH range of the produced lipase is 8.5-10.0, the optimal operative temperature range is 30-40 DEG C, and the residual enzyme activity is 80% after the temperature is maintained at 20 DEG C for 1 day. The lipase has obvious low temperature resistance and alkali resistance and can be widely applied to industrial production.

Description

Technical field: [0001] The invention belongs to the technical field of microbes, and in particular relates to a lipase and its fermentation production method and application. Background technique: [0002] Lipase (Triacylglycerol acylhydrolases, EC 3.1.1.3) is a general term for a class of enzymes that can hydrolyze triglycerides at the oil-water interface. Its natural substrates are generally water-insoluble long-chain fatty acid acyl esters. [0003] Lipases are mainly derived from plants, animals, and microorganisms. Among them, microbial lipases are widely found in bacteria, yeasts, and molds. Wide range of action temperature, action pH and substrate specificity, it can catalyze the hydrolysis, alcoholysis, acidolysis, transesterification and synthesis of ester compounds without coenzymes, with mild catalytic conditions, low energy consumption, and side effects The product is less, and it has the characteristics of high efficiency, high selectivity, and environmental ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/20C12R1/38
CPCC12N9/20C12Y301/01003
Inventor 刘文龙王兴吉郭庆文贾仁洁曹世源
Owner SHANDONG LONGKETE ENZYME PREPARATION
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