Asp305Gly mutation improved the activity and stability of the styrene monooxygenase for efficient epoxide production in Pseudomonas putida KT2440

Table 2 Substrate conversion and enantiomeric excess for the bioepoxidation of styrene, 3,3-dimethyl-1-butene, 3-allyl chloride and 2-chlorostyrene using the recombinant cells of Pseudomonas putida KT2440 (wild type and mutants) of SMO

The culture conditions are as follows: the single colonies were grown over 24 h at 30 °C in EM medium containing kanamycin (50 μg/mL). One milliliter of the culture was then inoculated into 100 mL of EM medium, then cultured at 30 °C for 24 h to induce protein expression. The cells were harvested by centrifuging at 8000×g for 8 min at 4 °C. The whole cell biotransformation with Pseudomonas putida KT2440/pJB861-styAB_D305X-fdh (SMO mutants, X = V, A, G) and Pseudomonas putida KT2440/pJB861-styAB-fdh (wild type SMO) were carried out in 50 mL flasks containing 10 mL of 200 mM KP buffer (pH 8.0) with 26 mM of one substrate, dry cell weight of 1.0 g, and 50% (v/v) hexadecane, incubated at 30 °C and 220 rpm on a rotatory shaker for 8 h. The product formation was determined by HPLC and the conversion was determined for the initial 30 min, as mentioned in “Materials and methods”

ISSN: 1475-2859