Lakhasly

A starter culture was inoculated from a single colony in 5 mL LB liquid medium supplemented with ampicillin (100 ug mL-1) and was incubated at 37 ?C and 180 rpm for 6 h. The whole starter culture was transferred to 30 mL LB containing 100 ug mL-1 ampicillin and cultivated at 37 ?C and 180 rpm for 16 h on an orbital shaker (Multitron, Infors, Switzerland).This cultivation protocol yields about 8 g L-1 cells (cell wet weight2.3.4.5.

A starter culture was inoculated from a single colony in 5 mL LB liquid medium supplemented with ampicillin (100 µg mL−1) and was incubated at 37 °C and 180 rpm for 6 h. The whole starter culture was transferred to 30 mL LB containing 100 µg mL−1 ampicillin and cultivated at 37 °C and 180 rpm for 16 h on an orbital shaker (Multitron, Infors, Switzerland).

The main cultures were inoculated with 6% (v/v) preculture. The preculture (30 mL) was added to 500 mL TB medium (5 g L−1 glycerol, 12 g L−1 tryptone, 24 g L−1 yeast extract, 0.17 M KH2PO4, 0.72 M K2HPO4), supplemented with 100 µg mL−1 ampicillin and incubated at 37 °C and 180 rpm until OD600=0.6–0.8 was reached.

Enzyme expression was induced with 1 mM IPTG and the cultures were incubated for 20 h at 25 °C and 180 rpm.

Cells were harvested by centrifugation (6000 x g, 20 min, 4 °C, Beckman Coulter, USA).

This cultivation protocol yields about 8 g L−1 cells (cell wet weight