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Streptococcus lactis peptide biological titration method

A technology of biological titer and nisin, which is applied in the field of determination of biological titer of nisin, can solve the problems of complex operation process and long time, and achieve the effect of reducing measurement time, simple operation and easy mastery

Inactive Publication Date: 2009-04-15
天津康益生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a fast method for measuring nisin biological potency in order to solve the complicated and time-consuming operation process of using agar diffusion method to measure nisin biological potency

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1: Fermentation fluid nisin biological potency

[0022] (1) Activate the detection bacterium-Sarcinia luteus in the solid slant detection medium prepared by hydrolyzed casein 1%, beef extract 0.5%, yeast powder 0.1%, NaCl 0.5%, agar powder 1.5%, culture 16- Store at 4°C for 24 hours and use within 3 days.

[0023] (2) Preparation of bacterial suspension: sterilize with distilled water at 120° C. for 20 minutes, and then insert the detection bacteria—Sarcinia luteus, so that the absorbance value of the bacterial suspension is 0.2.

[0024] (3) The nisin standard substance is made into 100 milliliters each of 6.66u / ml and 3.33u / ml with a pH of 2.5 hydrochloric acid aqueous solution, and then dilutes 1 The biological potency determination absorbance values ​​of the samples after doubling were 0.280 and 0.304, thus the determined standard curve was A=-0.0797InY+0.202.

[0025] (4) Preparation of detection culture medium: the formula of detection culture medium ...

Embodiment 2

[0029] Embodiment 2: salting out waste liquid nisin biological potency

[0030] (1) Activate the detection bacterium-Sarcinia luteus in the solid slant detection medium prepared by hydrolyzed casein 1%, beef extract 0.5%, yeast powder 0.1%, NaCl 0.5%, agar powder 1.5%, culture 16- Store at 4°C for 24 hours and use within 3 days.

[0031] (2) Preparation of bacterial suspension: sterilize with distilled water at 120° C. for 20 minutes, and then insert the detection bacteria—Sarcinia luteus, so that the absorbance value of the bacterial suspension is 0.1.

[0032] (3) the nisin standard substance is made into the hydrochloric acid aqueous solution of 2.5 with pH and is respectively 100 milliliters of biological potency of 300 and 600; The absorbance value was determined by valency, and the determined standard curve was A=-0.0797InY+0.202.

[0033] (4) Preparation of detection culture medium: the formula of detection culture medium is hydrolyzed casein 1%, beef extract 0.5%, ye...

Embodiment 3

[0037] Example 3: Biological potency of product nisin

[0038] (1) Activate the detection bacterium-Sarcinia luteus in the solid slant detection medium prepared by hydrolyzed casein 1%, beef extract 0.5%, yeast powder 0.1%, NaCl 0.5%, agar powder 1.5%, culture 16- Store at 4°C for 24 hours and use within 3 days.

[0039] (2) Preparation of bacterial suspension: sterilize with distilled water at 120° C. for 20 minutes, and then insert the detection bacteria—Sarcinia luteus, so that the absorbance value of the bacterial suspension is 0.1.

[0040] (3) The nisin standard substance is made into the hydrochloric acid aqueous solution of 2.5 with pH and is respectively 100 milliliters of biological potency of 300 and 600, according to the dilution sample corresponding to these two nisin standard solutions and the biological potency of the sample after diluting 1 times again The absorbance value was determined by valency, and the determined standard curve was A=-0.0797InY+0.202.

...

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Abstract

The invention is concerned with the determination method for the Nisin Biological value that belongs to the microbe determination method. It is: ensures the standard curve A=aInY+b; examines the substrate for activation of the bacterium - Micrococcus luteus; prepares bacterium suspension, the bacterium suspension cultivates in the testing substrate, uses the 752 spectrophotometer under the 600nm wavelength blank examination substrate as the consultation, determines the cultivated testing suspension absorption-light value A, computes the Biological value of the nisin sample according to the absorption-light value and the Nisin Biological value. The invention can short the determination time form 24 hours to 5 hours.

Description

technical field [0001] The invention relates to a measurement method including microorganisms, in particular to a method for determining the biological titer of nisin in a fermented liquid or a product by using a spectrophotometric method. Background technique [0002] Nisin is an effective biological preservative produced by microbial fermentation. It has been widely used in more than 50 countries for the preservation of milk and dairy products, fermented beverages, canned foods, meat and meat products. [0003] Currently, nisin is usually produced by microbial fermentation. The biological potency of nisin in the fermentation process, the biological potency of the product nisin and the biological potency of the compound preservative containing nisin are all measured by the agar diffusion method. The principle is to use the detection bacteria—Sarcina lutea (Sarcina lutea) to grow on the surface of the agar to display the inhibition zone of nisin to reflect the biological po...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/31C12Q1/02
Inventor 吴兆亮于广和胡国武
Owner 天津康益生物工程有限公司
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