Preparation of nitrite reductase and method for preparing nitrite reductase preparation

A technology of reductase and nitrous acid, which is applied in the field of preparation of biological enzymes and enzyme preparations, achieves the effects of short production cycle, reduction of human diseases, and promotion of quality improvement

Inactive Publication Date: 2008-07-30
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, an ideal substance that can completely replace nitrite has not been found so far. Count

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] 1. Culture Medium Preparation

[0031] (1) Formula of slant seed culture medium: 1-2wt% of nutrient broth powder, 1.8-2wt% of agar, pH6.5-7.5.

[0032] (2) Fermentation medium formula: glucose 1-2wt%, peptone 0.1wt%, sodium chloride 0.1-0.5wt%, dipotassium hydrogen phosphate 0.1-0.3wt%, potassium dihydrogen phosphate 0.05-0.2wt%, MgSO 4 ·7H 2 O 0.02-0.06wt%, MnSO 4 ·H 2 O 0.001-0.003wt%, CaCl 2 2H 2 O 0.002-0.004wt%, pH6.5-7.5.

[0033] 2. Dissolve the lyophilized powder of Bacillus megaterium in physiological saline at a weight ratio of 1:4, pick a ring with an inoculation loop, inoculate it on the slant seed medium, and cultivate it at 37°C for 20 hours to obtain an activated slant; ferment with 10mL The culture medium washes the activated slant bacteria under sterile conditions, puts it into a 250mL shake flask, fills each bottle with 100mL seed medium, and inoculates 2-4 seeds on each slant, at 25-30°C, with a rotation speed of 100-240r / min Cultivate at a con...

Embodiment 2

[0042] 1. Obtain the cell preparation

[0043] (1) Formula of slant seed medium: glucose 15.0g, NaNO 2 1.0g, K 2 HPO 4 1.35g, KH 2 PO 4 0.70g, MgSO 4 ·7H 2 O 0.20g, agar 20.0g, dilute to 1L, adjust the pH to 7.0, and sterilize at 121°C for 20min.

[0044] (2). Fermentation medium formula: glucose 15.0, peptone 2.0g, Na 2 HPO 4 1.35g, KH 2 PO 4 0.70g, MgSO 4 ·7H 2 O 0.10g, NaCl 1.0g, CaCl 2 2H 2 O 0.02g, MnSO 4 ·H 2 O 0.01g, NaNO 2 , 0.138g, dilute to 1L, adjust the pH to 7.0, and sterilize at 121°C for 20min.

[0045] 2. Dissolve the lyophilized powder of Bacillus megaterium in physiological saline at a weight ratio of 1:2, pick a ring with an inoculation loop, inoculate it on the slant medium, and culture it at 37° C. for 20 hours to obtain an activated slant. Then use the inoculation loop to pick 3-4 rings and inoculate them in 250mL shake flasks, each bottle contains 50mL of seed culture medium, and culture at 30-35°C for 24 hours at a constant tempera...

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PUM

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Abstract

The invention discloses a preparation method of preparing nitrite reductase and preparing the preparation of the nitrite reductase, which belongs to the preparing technology field of biologic enzyme and enzyme preparation. Bacillus megatherium which can produce nitrite reductase is activated and prepared into seed, the obtained seed culture fluid is inoculate into a liquid ferment substrate by 1-10 percent of inoculation dosage; the initial pH value is 5.0-7.5, at 25-40 DEG C and 100-240r/min of rotation speed, after 20-36 hours of constant temperature cultivation, bacterial suspension is produced. Re-suspending the bacterial suspension by phosphate buffer can produce thalli. After the thalli is smashed by ultrasonic, the supernatant fluid is frozen and centrifugated, and then the rough enzyme solution is obtained. Nitrite reductases of different consistences and purities can be obtained after the rough enzyme solution is separated and purified. Liquid enzyme preparation can be prepared after enzyme protecting solute is added in the nitrite reductase. The enzyme activity of nitrite reductase preparation produced by using the invention is 4000-8000U/mL. The preparing method is simple, the production period is short, and public food safety can be maintained.

Description

technical field [0001] The invention belongs to the technical field of preparation of biological enzymes and enzyme preparations, and more specifically, adopts fermentation technology to prepare a nitrite reductase capable of reducing nitrite and a preparation method of a nitrite reductase preparation. Background technique [0002] Nitrite, as a food additive allowed by the National Food Sanitation Law, is widely used in meat processing. It reacts with myoglobin to produce nitrosomyoglobin, which is pink, giving meat an attractive color, that is, color development; at the same time, it can inhibit the reproduction of microorganisms, especially the inhibition of Clostridium botulinum Propagation, this bacterium can produce botulinum toxin, cause food poisoning, that is, antiseptic effect; it can also make meat products elastic, taste good, eliminate the peculiar smell of raw meat, and improve product quality. However, nitrite is also recognized as a carcinogen, which has gre...

Claims

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Application Information

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IPC IPC(8): C12N9/02A23L1/314C12R1/11A23L13/40
Inventor 刘萍孙君社郑怀忠马鹏飞
Owner CHINA AGRI UNIV
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