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Method for producing renninum with red yeast liquid state fermentation

A technology of liquid fermentation and red yeast rice, which is applied in the field of bioengineering and enzyme preparation, can solve the problems of short microbial growth cycle, uncoordinated development, and inability to meet the needs of production, and achieve simple operation, low cost, and improved vitality. Effect

Inactive Publication Date: 2009-08-19
QILU UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because rennet is a key enzyme in cheese production, along with the continuous development of the cheese industry in the world, the method of slaughtering 50 million calves every year to obtain rennet still cannot meet the needs of production, and is also extremely incompatible with the development of modern industry uncoordinated
Therefore, researchers are constantly looking for new sources of rennet, mainly including animal rennet, plant rennet and microbial source rennet, but these sources of rennet all have certain limitation
Animal-based rennet is source-restricted, and cheese made from animal-based rennet does not meet the needs of vegans
Although plant-based rennet has a wide range of sources, it is difficult to develop due to constraints such as time, region, long growth cycle, and high proteolytic activity.
The use of microorganisms to produce rennet is the most promising development direction at present. The growth cycle of microorganisms is short, the output is large, and the limitation of climate, region and time is small. The cost of producing rennet with it is low, the enzyme extraction is convenient, and the economic benefit is high. , but there is also the problem of microorganisms producing other non-enzymatic metabolites

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Optimization of culture medium

[0034] 1) Effect of carbon source on chymosin activity

[0035] Glucose, sucrose, wheat bran, soluble starch and maltose were selected as carbon sources for experiments. The concentration of the carbon source is 50g / L, and other proportions of the medium are shown in the basic fermentation medium. When wheat bran was used as the carbon source for fermentation, the milk-clotting activity of chymosin was much higher than that of other carbon sources, indicating that wheat bran probably not only contributed to the growth of fermenting microorganisms, but more importantly It plays a positive role in the synthesis of the secondary metabolite of rennet. The influence of wheat bran may be that the composition of wheat bran is complex, and the organic or inorganic components produced are also more. In terms of fermentation, the nutritional components are more complete, and it is more conducive to the generation of secondary metabolites such as...

Embodiment 2

[0056] A method for preparing chymosin by liquid state fermentation of red yeast rice, the steps are as follows:

[0057] 1) Red yeast rice is crushed into powder with a fineness of 400 meshes. Inoculate the red yeast rice powder into PDA slant medium and culture at a constant temperature of 28°C for 90 hours to obtain purple-red Monascus; Monascus is purple-red and grows on red yeast rice around.

[0058] 2) Inoculate the red Monascus into the basic fermentation medium, and incubate at a constant temperature of 28° C. for 12 hours to carry out expanded cultivation;

[0059] The components of the basic fermentation medium are as follows, all in percent by weight:

[0060] Glucose is 1.5%; peptone is 0.5%; KH 2 PO 4 0.3%; K 2 HPO 4 0.3%; yeast powder 0.2%; MgSO 4 ·7H 2 O is 0.04%; ZnSO 4 ·7H 2 O is 0.03%, the balance is distilled water, and the pH is 5.6.

[0061] 3) Add the monascus after step 2) to the sterilized liquid culture medium by 5% (volume ratio) inoculum a...

Embodiment 3

[0070] A method for preparing chymosin by liquid state fermentation of red yeast rice, the steps are as follows:

[0071] 1) Red yeast rice is crushed into powder with a fineness of 200 meshes. Inoculate the red yeast rice powder into the PDA slant medium and culture at a constant temperature of 30°C for 96 hours to obtain purple-red Monascus; Monascus is purple-red and grows on red yeast rice around.

[0072] 2) Inoculate the red Monascus into the basic fermentation medium, and culture at a constant temperature of 30° C. for 12 hours to expand the culture;

[0073] The components of the basic fermentation medium are as follows, all in percent by weight:

[0074] Glucose is 1.5%; peptone is 0.4%; KH 2 PO 4 0.3%; K 2 HPO 4 0.4%; yeast powder 0.15%; MgSO 4 ·7H 2 O is 0.05%; ZnSO 4 ·7H 2 O is 0.02%, the balance is distilled water, and the pH is 6.0.

[0075] 3) Add the monascus after the expanded cultivation in step 2) to the sterilized liquid culture medium by 5% (volu...

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Abstract

The invention relates to a method for producing renninum by liquid state fermentation of red yeast rice, belonging to the technical field of biological engineering and enzyme preparation. The method comprises the following steps: liquid state fermentation of the red yeast rice; precipitation of organic solvent; low temperature centrifugation; vacuum freeze drying; Sephadex G-75 gel column chromatography; and DEAE-cellulose 52 chromatography. By the method, high-activity renninum can be prepared by raw materials with low cost so as to greatly solve the problem of deficient source of the renninum.

Description

【Technical field】 [0001] The invention relates to a method for producing chymosin by liquid-state fermentation of red yeast rice, and belongs to the technical fields of bioengineering and enzyme preparation. 【Background technique】 [0002] Chymosin (EC3.4.23.4) is an aspartic protease found in the abomasum of unweaned calves, and its main biological function is limited shearing of casein in κ-casein The peptide bond connected by Phe105-Met106 leads to milk coagulation, so it is widely used in cheese manufacturing and becomes an important enzyme preparation in the food field, and its output value accounts for 15% of the world's enzyme preparation. [0003] The traditional preparation method of rennet is to slaughter a calf 10 to 30 days old, and extract the rennet from its fourth stomach with salt. Because rennet is a key enzyme in cheese production, along with the continuous development of the cheese industry in the world, the method of slaughtering 50 million calves every ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/64C12R1/66
Inventor 王成忠于功明范素琴杜爱莲
Owner QILU UNIV OF TECH
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