Method for synthesizing L-fructose through whole-cell transformation
A whole-cell transformation and fructose technology, applied in the biological field, can solve the problems of difficult separation and purification of products, low conversion rate, etc., and achieve the effects of low price of raw materials, improved conversion rate, and reduced cost.
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Embodiment 1
[0020] A method utilizing whole cells to transform and synthesize L-fructose, using glycerol as a substrate (glycerol concentration is 500mM), adding L-glyceraldehyde (addition amount is 40mM), pyrophosphate (addition amount is 80mM) and recombinant Escherichia coli After the induced wet bacteria (the final concentration of the wet bacteria reaction system is 100g / L), adjust the pH to 7.0, control the reaction temperature to 30°C, and catalyze the reaction for 12 hours (the above-mentioned entire reaction system is 10mL). After the end, the supernatant was collected by centrifugation for 10 minutes, and the solution containing L-fructose was obtained.
[0021] Wherein the recombinant Escherichia coli is a recombinant Escherichia coli simultaneously expressing acid phosphatase PhoN, phosphoglycerol oxidase GPO and L-rhamnosan-1-phosphate aldolase RhaD.
[0022] Preparation of wet cells:
[0023] 1) Construction of the recombinant plasmid pET28a-rhaD: Using the genome of Escher...
Embodiment 2
[0033] The difference from Example 1 is that the reaction temperature is 25°C.
Embodiment 3
[0035] The difference from Example 1 is that the reaction temperature is 45°C.
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