新菌株Devosia sp.JM3中DON脱氢酶的基因组挖掘和酶学性质表征OA

The experiment aimed to mine the key detoxifying dehydrogenase for deoxynivalenol(DON)from the genome of the novel strain Devosia sp.JM3 and characterize its enzymatic properties.Firstly,bioinformatics software was used to perform genome assembly and gene prediction of the novel strain Devosia sp.JM3,and genome mining of the key DON detoxification dehydrogenase was completed through sequence alignment.Secondly,the enzyme was analyzed and validated for DON detoxification efficiency through molecular phylogenetic tree construction,sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE),and high-performance liquid chromatography(HPLC)detection.Finally,the effects of four factors,namely temperature,pH value,metal ions,and initial DON concentration,on the enzyme activity were investigated.The results showed that the target protein mined was completely conserved with the DON dehydrogenase A(DepA),exhibited high expression at 64 kDa,and could degrade 100 mg/L DON to 1.9%of its original concentration within 24 hours.Under the conditions of 35℃,pH value of 7.5,initial substrate concentration of 100 mg/L,and the presence of 1 mmol/L calcium ions(Ca2+),the enzyme achieved the highest conversion rate of DON.The study shows that the DON dehydrogenase mined from the genome of Devosia sp.JM3 is DepA,and its enzymatic properties are influenced by temperature,pH value,metal ions,and initial substrate concentration.