Method for fermentation production of lactase by aspergillus oryzae and lactase produced by method

A technology of lactase and Aspergillus oryzae, applied in the field of bioengineering, can solve the problems of low unit yield, high cost, and poor heat resistance of lactase, and achieve the effects of expanding the scope of industrial applications, reducing fermentation costs, and increasing fermentation capacity

Active Publication Date: 2019-02-01
SHANDONG LONGKETE ENZYME PREPARATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A lot of work has been done abroad since the 1960s and at home since the 1980s, but there is still a big gap between domestic and foreign countries. Lactase has not been widely used in China's food and dairy industries. The main reason is that lactase The unit yield of Lactase is relatively low. Most of the lactases sold on the market are intracellular enzymes, which need to be broken into cells to obtain. The extraction and purification process is complicated, the cost is high, the action temperature of the enzyme is low, and the heat resistance is poor. It is easy to be infected with bacteria in industrial production. Narrow scope of application

Method used

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  • Method for fermentation production of lactase by aspergillus oryzae and lactase produced by method
  • Method for fermentation production of lactase by aspergillus oryzae and lactase produced by method
  • Method for fermentation production of lactase by aspergillus oryzae and lactase produced by method

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Experimental program
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Effect test

Embodiment 1

[0036] The mutagenesis selection of embodiment 1 Aspergillus oryzae CA-08

[0037] Take the fresh slant of the two starting strains AJ2010, wash the spores on the fresh slant with 10mL 0.1% Tween-60, transfer it into a triangular flask filled with glass beads, shake it on the shaker for 50min, and then wash it with 4 layers Filter with lens paper, and perform gradient dilution, count on a hemocytometer and adjust the spore concentration to 6×10 6 cells / mL as the starting bacterial suspension.

[0038] Place a test tube containing an appropriate amount of spore suspension on an irradiation plane with a certain dose rate, and use Co 60 Radiation mutagenesis was carried out with radiation doses of 50,000, 60,000, 70,000, 80,000, 90,000, 100,000, and 110,000 roentgens respectively. After treatment, gradient separation was carried out and cultured at 30°C for 60 hours. The lethality rate varies with Co 60The radiation dose increases, and the positive mutation rate increases with...

Embodiment 2

[0049] Embodiment 2 Aspergillus oryzae CA-08 fermentation produces enzyme and the extraction method of the produced lactase

[0050] The fermentative enzyme production method of aspergillus oryzae CA-08 mainly comprises the following steps:

[0051] Slant culture: Pick a ring of Aspergillus oryzae CA-08 and inoculate it on a solid slant medium, and culture at a constant temperature of 30°C for 36 hours to obtain first-grade seeds;

[0052] Shake flask culture: Take a ring of the first-class seeds and insert them into the seed medium, and cultivate them for 48 hours at a constant temperature of 30°C and a shaker speed of 200r / min to obtain the second-class seed liquid;

[0053] Seed tank cultivation: put the secondary seed solution into the seed tank culture medium at a ratio of 15% (v / v) of the inoculation amount, and cultivate for 45 hours at a constant temperature of 30°C and a rotational speed of 200r / min;

[0054] Fermentation tank culture: The seed liquid in the seed tan...

Embodiment 3

[0064] Embodiment 3 Aspergillus oryzae CA-08 fermentation performance verification

[0065] Carry out the 50L fermenter verification experiment according to the fermentative enzyme production method of embodiment 2 Aspergillus oryzae CA-08, the fermentation period is 105h, its 7 batches of fermentation enzyme production situation, the average enzyme production level is 100762U / mL, the following table shows that the bacterial strain is not only high yield Lactase and its fermentation performance and the enzymatic activity of the lactase produced by it all have remarkable stability.

[0066] Fermentation enzyme production situation of table 7 batches of high-yielding lactase strains

[0067]

[0068]

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Abstract

The invention discloses a method for fermentation production of lactase by aspergillus oryzae and the lactase produced by the method, and belongs to the bioengineering technical field. The strain is specifically aspergillus oryzae CA-08 and has the preservation number of CGMCC No.14137. The strain is deeply fermented in a liquid 50 L tank, is supplemented with materials and is subjected to fermentation culture for 105 h, and the enzyme production average level is 100762 U / mL. The established method for liquid fermentation of the lactase has the optimum pH range for production of the lactase of4.0-7.0, the optimum action temperature is 80 DEG C, the residual enzyme activity is 89.5% after heat preservation is carried out for 1 h at the temperature of 80 DEG C, and the residual enzyme activity is 58.9% after heat preservation is carried out for 2 h at the temperature of 80 DEG C. The lactase has good heat resistance and can be widely applied in industrialized production.

Description

Technical field: [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a method for producing lactase by fermenting aspergillus oryzae and the produced lactase. Background technique: [0002] Lactase is a tasteless, odorless, light brown liquid after dissolution, non-toxic and no side effects of biological enzyme preparation, the enzyme can be used to degrade lactose into galactose and glucose, and also has the transfer function of galactoside. It is also an important nutrient in mammalian milk. It can catalyze the hydrolysis of lactose into galactose and glucose, and is mostly used in the dairy industry. Lactase activity in mammals typically decreases physiologically with age, so lactase deficiency is a widespread worldwide problem. [0003] Lactase exists in plants such as almonds and apricots and in the small intestine of young mammals. Microorganisms such as bacteria, molds and yeasts can also produce lactase after fermenta...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/38C12R1/69
CPCC12N9/2471C12Y302/01023
Inventor 王克芬张杰郭庆文王兴吉刘文龙曹世源
Owner SHANDONG LONGKETE ENZYME PREPARATION
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