Reaktion #731768

Reaktionsgleichung

MgCl2

NaCl

CaCO3

carbenicillin

IPTG

→

L-ascorbic acid

Reaktionsbedingungen

Aufarbeitung

1. 1
   workup.WAITto be cultivated at 230 rpm at 25° C. for 3 h
2. 2
   Waschenwashed twice with saline
3. 3
   Sonstigecell reaction with the reaction mixture (500 μl in 5 ml tube)
4. 4
   workup.WAITconducted at 30° C. for 15 h

Vorschrift

Five ml of one overnight culture of E. coli BL21 (DE3)/pET21b-SNDHaiHis in LB with carbenicillin 50 μg/ml was inoculated into 200 ml of the same medium. The cells were cultivated at 230 rpm at 37° C. for 2 h, then induced with 1 mM IPTG and continued to be cultivated at 230 rpm at 25° C. for 3 h. The resulting culture was centrifuged and washed twice with saline and the cell pellet was resuspended in 2 ml of water. The cells were used for resting cell reaction with the reaction mixture (500 μl in 5 ml tube) containing cells at OD600=10, 1% sorbosone monohydrate, 5 μM PQQ, 5 mM MgCl2, 0.3% NaCl, and 1% CaCO3 conducted at 30° C. for 15 h. 0.14 g/L of L-ascorbic acid was produced after incubation for 15 h. When the resting cell reaction was done with 1 μM PQQ (the other conditions were same as those described above), 0.05 g/L of L-ascorbic acid was produced after incubation for 3 h.

Quelle