Method for increasing phytoene dehydrogenase in vitro reaction rate

A technology of phytoene and lycopene, which is applied in the field of bioengineering, can solve problems such as slow reaction rate, and achieve the effects of reducing oxidative loss, improving accuracy, and improving enzymatic reaction rate.
CN102286593BInactive Publication Date: 2013-03-13SHANDONG UNIV
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
SHANDONG UNIV
Publication Date
2013-03-13
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention relates to a method for increasing water-organic double-phase enzymatic reaction rate, in particular relates to a method for increasing phytoene dehydrogenase in vitro reaction rate and belongs to the technical field of biological engineering. The method comprises the following steps of: adding phytoene acetone solution, glucose, glucose oxidase and hydrogenperoxidase into crushed coarse enzyme solution to ultrasonically mix uniformly; performing dark airtight oscillating reaction at the temperature of between 28 and 30 DEG C; adding methanol; performing warm bath at the temperature of between 55 and 60 DEG C for 15 to 20 minutes; adding petroleum ether with the boiling range of 30 to 60 DEG C to oscillate to extract reaction products; collecting the upper layer of a petroleum ether layer; and performing vacuum rotary evaporation to obtain the product. A phytoene dehydrogenase in vitro reaction system is optimized by utilizing the mechanical effects such as homogeneity, dispersion, emulsification and the like of ultrasonic wave, so probability of a substrate contacting the enzyme is improved, the enzymatic reaction rate is obviously increased, reaction time is shortened, oxidation loss of the product is reduced, and accuracy of data is improved.
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Description

technical field

[0001] The invention relates to a method for increasing the rate of water-organic two-phase enzymatic reaction, in particular to a method for increasing the in vitro reaction rate of phytoene dehydrogenase, belonging to the technical field of bioengineering. Background of the invention

[0002] Phytoene desaturase (phytoene desaturase 1.14.99.-) is a key enzyme in the synthesis and metabolism of carotenoids, which can catalyze colorless phytoene to generate colored carotenoids. Enzymes are widely found in plants and microorganisms. According to the number of dehydrogenation, there are at most 6 kinds of products of phytoene dehydrogenase, which are hexahydrophytoene, ζ-carotene, streptoerythrin, lycopene, 3,4-dide Phytoene, 3,4,3',4'-tetradehydrolycopene, among which phytoene dehydrogenase with streptosporine and lycopene as products is the most common. Almost all carotenoids naturally occurring in nature are produced with streptosporin or lycopene as inter...

Claims

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