<15>N SIP (stable isotope probing) vibrio parahaemolyticus culture medium and cultivation method thereof

Abstract

The invention discloses a <15>N SIP (stable isotope probing) vibrio parahaemolyticus culture medium and a cultivation method thereof. A formula of the culture medium comprises components as follows: 1-5 g / L of a nitrogen source, 5-18 g / L of a carbon source, 0.5-2.0 g / L of KH2PO4, 0.6-2.0 g / L of K2HPO4, 0.1-1.0 g / L of MgSO4, 10-30 g / L of NaCl and 1L of distilled water, the pH is in a range from 7.0 to 7.5, and nitrogen atoms of the nitrogen source are labeled by adopting <15>N. The cultivation method of the <15>N SIP vibrio parahaemolyticus comprises steps as follows: 1), continuously and repeatedly cultivating the vibrio parahaemolyticus through transferring; 2), verifying the labeling efficiency of protein nitrogen atoms in labeled vibrio parahaemolyticus cells with LC-MS / MS (high-performance liquid chromatography-tandem mass spectrometry). The process is simple and reasonable, the culture medium has simple components and is low in cost, and the novel culture medium preparation method is provided for industrial preparation of the <15>N SIP vibrio parahaemolyticus culture medium.

Description

technical field [0001] The present invention relates to a 15 N stable isotope labeled Vibrio parahaemolyticus culture medium and its cultivation method. Background technique [0002] In the marine environment, most Vibrio species are the most common type of bacteria that maintain normal marine habitats, but some Vibrio species are harmful to humans and marine animals. In 1995, the sixth edition of the American Manual of Clinical Microbiology identified 12 types of pathogenic bacteria in Vibrio, the most important of which were Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus. FDA in the United States, NMKL in Europe, and the Ministry of Health in China have all formulated corresponding detection methods. Diseases caused by pathogenic Vibrio include gastroenteritis, wound infection, etc. For example, food poisoning caused by Vibrio parahaemolyticus is transmitted through improperly cooked seafood or salted products. The common ones are jellyfish, sea fish, s...

AI Technical Summary

Problems solved by technology

The former generally uses a medium containing heavy stable isotope essential amino acids to cultivate microorganisms, but no matter the method of microbial fermentation or organic chemical synthesis of stable isotope-labeled amino acids is used, the cost is high and the process is complicated; the latter adopts protein extraction. Functional groups (such as peptides) chemically react with stable isotope-labeled reagents, with many side reactions and complex processes

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