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Detection method for titer of nisin in fermentation liquor

A technology of nisin and detection method, which is applied in the field of improvement of the method for rapid batch detection of nisin titer, can solve the problems of poor accuracy of experimental results, slow growth, long experimental period and the like, and can meet the requirements of rapid detection. , improve accuracy and save time

Inactive Publication Date: 2016-04-06
石家庄新宇三阳实业有限公司
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  • Application Information

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Problems solved by technology

[0008] In the process of using spectrophotometry to detect the growth of indicator bacteria, it was found that Micrococcus luteus was cultured according to the instructions given by the China Industrial Microbiology Culture Collection Management Center. It took 1-2 days at 30°C to grow. Slow, resulting in long experiment period and poor accuracy of experiment results

Method used

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  • Detection method for titer of nisin in fermentation liquor
  • Detection method for titer of nisin in fermentation liquor
  • Detection method for titer of nisin in fermentation liquor

Examples

Experimental program
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preparation example Construction

[0021] Preparation of liquid basal medium: 5.0 g of peptone, 3.0 g of beef extract, 5.0 g of sodium chloride, and the above were dissolved in 1.0 L of distilled water, and the pH value was adjusted to 7.0.

[0022] Preparation of solid basal medium: 15.0 g of agar was added to the above liquid basal medium.

[0023] Use an inoculation loop to pick the indicator bacteria stored at -40°C and inoculate them into the solid basal medium, and culture them at 30°C for 24 hours. Then pick a single colony and inoculate it into 10ml of liquid basal medium, culture it in a shaker at 30°C and 120r / min for about 7 hours to obtain a bacterial suspension, which is used for the culture medium optimization and titer determination.

Embodiment 1

[0025] Into a 250ml Erlenmeyer flask containing 50ml of liquid basal medium, insert 500ul of bacterial suspension, culture at 30°C on a shaker at 120r / min, and take samples every half hour. The concentration of Micrococcus luteus bacteria solution was determined by turbidimetric method, that is, the absorbance value was measured at a wavelength of 600nm, and the liquid basal medium without bacterial suspension was used as a reference. Finally, the growth curve was drawn with time as the abscissa and OD value as the ordinate.

Embodiment 2

[0027] Add glucose 0.1%, K 2 HPO 4 0.1%, MgSO 4 ·7H 2 O0.01%, optimize the liquid basal medium; insert 500ul bacterial suspension, and cultivate at 30°C, 120r / min shaker. The concentration of Micrococcus luteus bacterium liquid is measured by turbidimetry, and the optimized liquid basal medium described in this embodiment without adding bacterial suspension is used as a reference, and its absorbance value is measured under the condition of wavelength 600nm after culturing for 5 hours is 0.913.

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Abstract

The invention discloses a detection method for titer of nisin in fermentation liquor. The spectrophotometric method is adopted, and the detection method specifically includes the steps of selecting an indicator, manufacturing a standard curve by measuring the OD value, indicating indicator growth, in fermentation liquor of a culture medium containing nisin of a certain concentration, conducting direct inoculation on the fermentation liquor, and the concentration is compared with the concentration of nisin in the fermentation liquor to be determined, wherein micrococcus luteus is selected as the indicator. By means of the design, system optimization is conducted on the culture medium of micrococcus luteus, the spectrophotometric method which can rapidly detect the improvement of the titer of Nisin is established on this basis, and therefore time is saved, accuracy is improved, and cost is reduced. The formula of the culture medium is optimized during detection, some nutritional ingredients are added on the basis of the basic culture medium, growth of the indicator is ensured, and the requirement for rapidly detecting a large number of samples is met.

Description

technical field [0001] The invention relates to the detection of nisin in fermented liquid, in particular to the improvement of a rapid batch detection method for nisin titer. Background technique [0002] Nisin (Nisin, referred to as Nisin) is a polypeptide substance extracted from the metabolites of Lactococcus lactis. It consists of 34 amino acids and can significantly inhibit the growth of Gram-positive bacteria. It can be used in milk and dairy products 1. Preservation and preservation of meat and meat products. It is a recognized safe and non-toxic natural biological food preservative and antibacterial agent. It is widely used in the food industry and has been approved by more than 60 countries. [0003] The initial determination method of Nisin quantitative analysis was the methylene blue reduction method suggested by CoxGA et al. in 1934, and later developed biological analysis methods for Nisin quantification, including tube dilution (TubeDilution), turbidity analys...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31
CPCG01N21/31
Inventor 张拴力扈士海赵地顺张桂彦王素新
Owner 石家庄新宇三阳实业有限公司
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