ABSTRACT
AIMS: This study aimed to improve the production of mutantioxidin, an antioxidant encoded by a biosynthetic gene cluster (mao) in Streptococcus mutans UA140, through a series of optimization methods. METHOD AND RESULTS: Through the construction of mao knockout strain S. mutans UA140∆mao, we identified mutantioxidin as the antioxidant encoded by mao and verified its antioxidant activity through a reactive oxygen species (ROS) tolerance assay. By optimizing the culture medium and fermentation time, 72 h of fermentation in chemically defined medium (CDM) medium was determined as the optimal fermentation conditions. Based on two promoters commonly used in Streptococcus (ldhp and xylS1p), eight promoter refactoring strains were constructed, nevertheless all showed impaired antioxidant production. In-frame deletion and complementation experiments demonstrated the positive regulatory role of mao1 and mao2, on mao. Afterward, the mao1 and mao2, overexpression strain S. mutans UA140/pDL278:: mao1mao2, were constructed, in which the production of mutantioxidin was improved significantly. CONCLUSIONS: In this study, through a combination of varied strategies such as optimization of fermentation conditions and overexpression of regulatory genes, production of mutantioxidin was increased by 10.5 times ultimately.
Subject(s)
Dental Caries , Streptococcus mutans , Humans , Streptococcus mutans/genetics , Antioxidants , Streptococcus , Promoter Regions, Genetic , Monoamine Oxidase/genetics , Biofilms , Dental Caries/prevention & controlABSTRACT
Introducing the theory of fuzzy set, mathematical morphology and computerized mask fast scanning, we developed the TOOTH.SCA software and method to analyze the effect of fluoride (NaF) on ore content of human tooth enamel automatically and quantitatively. And we obtained some characteristic parameters, such as the depth, the type and the demineralized content of every scathing layer of dental caries. The smallest scale of mask scanning is 0.1 microm x 0.1 microm and the time required to analyze a sample is only 12 s. The applied software and method we built play an important role to the research on the mechanism of pathological changes of teeth and preventing dental caries.