une cuve spectrophotométrique (1mL), sont introduits : -800 µL de tampon glycyl-glycine 0,1 M pH 7,5 -20 µL de NADH,H + (10 mg.mL -1 ), -5 µL de ThDP (10 mg.mL -1 ), -10 µL de MgCl 2 (10 mg.mL -1 ), -100 µL de L-érythrulose (120 mg.mL -1 ), -50 µL de D-ribose-5-phosphate (50 mg, mL -1 ), -5 µL d'ADH (5000 U.mL -1 ), -10 µL de TK (V TK ) ,
Après 5 minutes de centrifugation à 14000 rpm, 800 µL d'acétonitrile sont ajouté à 200 µL de surnageant pour obtenir un échantillon destiné à l'injection en ,
les culots sont repris doucement avec 200 mL, 100 mL, 20 mL d'eau stérile osmosée (stocké à 4 °C) et enfin avec 2 mL d'un solution glacée de glycérol à 20 % (les cellules sont centrifugées entre chaque reprise) Les cellules ainsi préparées peuvent être utilisées fraichement préparées ,
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