High-throughput rapid screening method for L-threonine transaldolase mutant
A screening method and threonine technology, applied in the fields of biochemistry and enzyme engineering, can solve problems such as low efficiency, time-consuming and laborious, and achieve the effects of good repeatability, high sensitivity and accuracy, and easy popularization and application.
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[0030] Preferred embodiments of the present invention are provided below to help further understanding of the present invention. Those skilled in the art should understand that the descriptions of the embodiments of the present invention are only exemplary, and are not intended to limit the solution of the present invention.
[0031] Step 1. Reagent preparation
[0032] L-threonine transaldolase whole cell reaction solution: Randomly pick mutant single clones from the L-threonine transaldolase mutation library, inoculate into 48-well deep-well plates (0.3mL LB liquid medium, containing 50μg / mL kanamycin) at 37°C, 180rpm overnight. The next day, transfer 50 μL of cell culture solution to another 48-well deep-well plate (containing 950 μL LB liquid medium, 50 μg / mL kanamycin), and culture at 37° C. for 2-3 hours with shaking at 200 rpm. Add IPTG to a final concentration of 0.1 mM, and stop culturing after induction at 30°C for 8 hours. Centrifuge at 4000 rpm for 10 min at 4°...
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