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A kind of preparation method of composite enzyme for extracting vegetable oil by aqueous enzymatic method

A vegetable oil and compound enzyme technology, which is applied to biochemical equipment and methods, hydrolytic enzymes, enzymes, etc., can solve the problems of high cost and complicated compound enzyme production process, and achieve short production cycle, rich enzyme system and neutral protease The effect of high vitality

Active Publication Date: 2018-10-19
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

At present, the compound enzymes used in the aqueous enzymatic method are basically composed of several purebred enzyme preparations in a certain proportion. The production process of the compound enzymes prepared by this method is complicated and the cost is high.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] 1. Take a ring of Bacillus amyloliquefaciens Z16-1 from the bacterial slant and inoculate it into the nutrient agar medium, streak culture at 36°C for 42 hours, and select a well-growing single colony for seed liquid culture.

[0025] 2. Inoculate the activated strain into a broth medium (50mL broth medium in a 250mL Erlenmeyer flask), and culture in a shaking flask at 36°C and 240r / min for 26h to obtain a seed solution.

[0026] 3. Inoculate the seed solution into the fermentation medium (100mL fermentation medium in a 500mL Erlenmeyer flask) with an inoculation amount of 4% (v / v), and culture in a shaking flask at 36°C and 240r / min for 46h.

[0027] 4. Centrifuge the obtained fermented liquid at 6000r / min for 12min, and the supernatant is the compound enzyme enzyme liquid suitable for extracting camphor tree seed kernel oil by aqueous enzymatic method. In the enzyme liquid: the activity of neutral protease is 9027±2% U / mL, the activity of amylase is 15.6±2% U / mL, the ...

Embodiment 2

[0029] 1. Take a ring of Bacillus amyloliquefaciens Z16-1 from the bacterial slant and inoculate it into the solid medium, streak culture at 37°C for 36 hours, and select a well-growing single colony for seed liquid culture.

[0030] 2. Inoculate the activated strain into broth medium (50mL broth medium is packed in a 250mL Erlenmeyer flask), and culture in shake flask at 37°C and 220r / min for 24h to obtain seed liquid.

[0031] 3. Inoculate the seed liquid into the fermentation medium (100mL fermentation medium in a 500mL Erlenmeyer flask) with an inoculation amount of 3% (v / v), and culture in a shaker flask at 37°C and 220r / min for 44h.

[0032] 4. Centrifuge the obtained fermented liquid at 7000r / min for 10min, and the supernatant is the compound enzyme enzyme liquid suitable for extracting camphor tree seed kernel oil by aqueous enzymatic method. In the enzyme liquid: the activity of neutral protease is 8594±2% U / mL, the activity of amylase is 19±2% U / mL, the activity of g...

Embodiment 3

[0034] 1. Take a ring of Bacillus amyloliquefaciens Z16-1 from the bacterial slant and inoculate it into the solid medium, streak culture at 38°C for 24 hours, and select a well-growing single colony for seed liquid culture.

[0035] 2. Inoculate the activated strain into a broth medium (50mL broth medium in a 250mL Erlenmeyer flask), and culture in a shaking flask at 38°C and 200r / min for 22h to obtain a seed solution.

[0036] 3. Inoculate the seed liquid into the fermentation medium (100mL fermentation medium in a 500mL Erlenmeyer flask) with an inoculum amount of 2% (v / v), and culture in a shaker flask at 38°C and 200r / min for 42h.

[0037] 4. Centrifuge the obtained fermented liquid at 6000r / min for 12min, and the supernatant is the compound enzyme enzyme liquid suitable for extracting camphor tree seed kernel oil by aqueous enzymatic method. In the enzyme solution: the activity of neutral protease is 7411±2% U / mL, the activity of amylase is 29±2% U / mL, the activity of gl...

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Abstract

The invention discloses a preparation method of a complex enzyme for extracting vegetable fat through an aqueous enzymatic method. The method comprises the following steps: (1) inoculating preserved bacillus amyloliquefaciens Z16-1 into a nutrient agar medium, and activating and cultivating the bacillus amyloliquefaciens Z16-1 for 24-42h at the temperature of 36-38 DEG C; (2) selecting an activated strain from the nutrient agar medium, inoculating the activated strain into a broth medium, and performing shake-flask culture for 22-26h at the temperature of 36-38 DEG C and at the speed of 200-240r / min to prepare a seed solution; (3) adding the seed solution into fermented cultivation with the inoculation amount of 2-4 percent, performing shake-flask culture for 42-46h at the temperature of 36-38 DEG C and at the speed of 200-240r / min to prepare a fermentation solution, and centrifuging the fermentation solution for 8-12min at the speed of 6,000-8,000r / min to obtain a supernatant, wherein the supernatant is an enzyme solution of the complex enzyme. The preparation method has the advantages of simple production process, short production period, energy conservation, environmental protection and the like, an enzyme system of the obtained complex enzyme is rich, the activity of neutral protease is high, lipase activity is zero, and the extraction rate of fat and the extraction rate of protein hydrolysate in a process of extracting the vegetable fat through the aqueous enzymatic method can be effectively improved.

Description

technical field [0001] The invention belongs to the technical field of bioengineering fermentation, and in particular relates to a method for producing compound enzymes by fermentation of Bacillus amyloliquefaciens. Background technique [0002] Due to the deep demand for food safety, environmental protection and comprehensive utilization of oil resources, the technology of extracting vegetable oil by aqueous enzymatic method has become the research frontier at home and abroad. The principle of water enzymatic extraction of vegetable oil is: after the oil is mechanically crushed, biological enzymes are added to fully destroy the cell tissue of the oil to release the oil, and finally the vegetable oil and hydrolyzed protein are obtained by centrifugation. Biological enzymes (such as protease, cellulase, pectinase, dextranase, amylase, etc.) that can destroy the cell tissue of plant oils can be applied to the extraction of vegetable oil by aqueous enzymatic method, and the com...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/00C12N9/54C12N9/28C12N9/34C12N9/88C12N9/26C12N9/18C12N9/42
Inventor 曾哲灵王健宇肖彥骏曾诚余平文学方
Owner NANCHANG UNIV
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