Composition with enzymatic anti-inebriation effect and preparation method and application thereof
A technology of composition and function, which is applied in the field of enzyme-catalyzed anti-alcoholism composition and its preparation, can solve the problems of unreported, difficult to decompose alcohol, lack of alcohol, etc., and achieve the reduction of alcohol content, quick effect, and comprehensive prescription Effect
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Embodiment 1
[0043] Cloning (yeast), expression, purification of embodiment 1 alcohol dehydrogenase
[0044] 1. Plasmid construction. The ADH1B2 gene (corresponding NCBI accession number CN20036-1) on the commercial plasmid ADH1B2-pPIC9k (purchased from Jiangsu Hongxun Biotechnology Co., Ltd., containing human ADH1B2 gene and N-terminal-His×6 tag) Amplified by PCR (upstream primer ADH1B2-pPIC3.5k-F: aattattcgaaggatccATGGCCACCATGCACCACCACCATCACCA, downstream primer ADH1B2-pPIC3.5k-R: ccgccctagggaattcTCAAAACGTCAGGACGGTACG), after obtaining the target fragment (including ADH1B2 and His×6 tags), use homologous recombinase ⅡOne StepCloning Kit, the human alcohol dehydrogenase ADH1B2 gene and the His×6 tag were constructed between the BamHI and EcoR I restriction sites of the pPIC3.5k plasmid to obtain the expression vector ADH1B2-pPIC3.5k.
[0045] 2. Construction of a yeast expression system. The plasmid ADH1B2-pPIC3.5k obtained in step 1 was digested with Sac I fast enzyme and transformed in...
Embodiment 2
[0049] The cloning (yeast), expression and purification of acetaldehyde dehydrogenase are similar to the above method.
[0050] 1. Plasmid construction. The ALDH2 gene (corresponding NCBI accession number CNCN19443-1) on the commercial plasmid ALDH2-pPIC9k (purchased from Jiangsu Hongxun Biotechnology Co., Ltd., containing human ALDH2 gene and N-terminal-His×6 tag) Amplified by PCR technology (upstream primer ALDH2-pPIC3.5k-F: attattcgaaggatccATGGCCACCATGCACCACCACCATCACCA, downstream primer ALDH2-pPIC3.5k-R: ccgccctagggaattcTTAGGAGTTCTTTTGTGGGACCTT), after obtaining the target fragment (including ALDH2 gene and His×6 tag), use homologous recombination enzyme ⅡOneStep Cloning Kit, the human acetaldehyde dehydrogenase ALDH2 gene and His×6 tag were constructed between the BamH I and EcoR I restriction sites of the pPIC3.5k plasmid to obtain the expression vector ALDH2-pPIC3.5k.
[0051] 2. Construction of yeast expression system. The plasmid ALDH2-pPIC3.5k obtained in step 1 wa...
Embodiment 3
[0054] Preparation 1 of embodiment 3 enzyme catalyzed hangover composition
[0055] (1) Alcohol dehydrogenase 1g, acetaldehyde dehydrogenase 1g, alcohol dehydrogenase agonist dithiothreitol 1g, acetaldehyde dehydrogenase agonist methionine 1g, and the common prosthetic group of the two dehydrogenases Mix 1 g of nicotinamide adenine dinucleotide evenly to obtain the enzymatic hangover hangover composition I.
[0056] (2) Constant volume: Dissolve the hangover agent composition I raw material obtained in step (1) in distilled water to prepare a 5 mg / mL aqueous solution.
[0057] (3) The aqueous solution in the step (2) is sterilized by filtration through a microporous membrane with a pore size of 0.22 μm, filled in a clean, dry and sterilized container, and sealed with a cover to obtain a hangover-relief composition.
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