Preparation method and application of carotenoid 9,10' dioxygenase
A technology of carotene and dioxygenase, which is applied in the field of bioengineering, can solve the problems of high requirements on reaction conditions, low yield, and long reaction time of β-ionone, so as to achieve less reaction products, short reaction time, and optimized Effect of Reaction Conditions
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[0030] 1. Preparation of Enterobacter carotenoid 9, 10' dioxygenase
[0031] The preparation method of Enterobacteriaceae carotenoid 9, 10' dioxygenase, including strain, plasmid, enzyme and culture medium, primer design, PCR amplification and construction of recombinant plasmid, induced expression of gene and purification of protein. Specific steps are as follows:
[0032] (1) Strain, plasmid, enzyme and culture medium:
[0033] Escherichia coli ( Escherichia coli )DH5α, restriction endonuclease Nde I and Sal I. Pyrobest DNA polymerase, DNA ligation kit ver.2.1, bacterial genome DNA extraction kit ver.3.0 were purchased from TAKARA company; Escherichia coli Escherichia coli BL21-codonPlus(DE3)-RIL was purchased from Novagen’s LB medium: each liter contained 10 g of Tryptone, 5 g of Yeast extract, and 10 g of NaCl; the concentration of ampicillin was 100 mg / L, and the concentration of chloramphenicol was 34 mg / L ; Enterobacter hallii ( Enterobacter hormaechei ) is...
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