Detection method of myrosinase activity in probiotics

Abstract

The invention discloses a method for detecting activity of myrosinase in probiotics. According to the condition that myrosinase can be an induced enzyme, the reaction substrate sinigrin of myrosinase is utilized to induce the strain, and the activity of the myrosinase is then detected. The method comprises the following steps: carrying out primary screening by using sinigrin instead of glucose to primarily judge whether the strain has the capacity for decomposing sinigrin; inducing the strain by using the sinigrin so that the strain generates metabolism channel activation of myrosinase, adding the complete strain into a sinigrin solution with a certain concentration to perform metabolism for some time, centrifugating to remove the strain, and determining the 340nm absorbance of the supernatant, wherein the result indicates that the absorbance value at this time is obviously changed as compared with the absorbance of the sinigrin solution with the same concentration into which the strain is not added. Therefore, the strain induction method for determining activity of myrosinase in probiotics is feasible. The method is simple to operate and short in detection period.

Description

technical field [0001] The invention relates to a method for detecting the activity of probiotic myrosinase. Background technique [0002] The scientific name of myrosinase is β-glucosinosidase, also known as β-glucosinolate hydrolase, which is mainly found in cruciferous plants and is the only β-glucosinolate found in nature so far. Glycoside hydrolase. Myrosinase has a variety of isoenzymes, which exist in different cruciferous plant tissues. Myrosinase purified from rapeseed and white mustard is a dimeric protein with a molecular mass of about 135kD-150kD. Myrosinase rarely exists in free form, but instead forms protein complexes with coenzymes such as myrosinase-binding protein or myrosinase assisting protein. In general, myrosinase in cruciferous plants will form a relatively independent glucosinolate-myrosinase system with glucosinolate glucoside. Only when the plant tissue is damaged can the two come into contact with each other and react to generate toxic Thiocya...

AI Technical Summary

Problems solved by technology

However, the above studies only proved that intestinal microorganisms have myrosinase activity by measuring the generation of the decomposition product isothiocyanate, and did not further purify and analyze myrosinase or measure its enzyme activity, and the above studies mostly used Mass spectrometry or chromatography, which requires high experimental equipment and experimental costs