3.1.1.84 A199S/F227A/S287G/A328W/E441D the mutant shows 1730fold improved (-)-cocaine-hydrolyzing activity compared to the wild type enzyme 716278 3.1.1.84 A199S/F227A/S287G/A328W/Y332G the mutant has a 2020fold improved catalytic efficiency against (-)-cocaine compared to the wild type enzyme 714226 3.1.1.84 A199S/F227A/S287G/A328W/Y332G/E441D the mutant shows 1390fold improved (-)-cocaine-hydrolyzing activity compared to the wild type enzyme 716278 3.1.1.84 A199S/F227A/S287G/A328W/Y332G/F329V the mutant has a 121fold improved catalytic efficiency against (-)-cocaine compared to the wild type enzyme 714226 3.1.1.84 A199S/F227I/S287G/A328W/Y332G the mutant has a 1170fold improved catalytic efficiency against (-)-cocaine compared to the wild type enzyme 714226 3.1.1.84 A199S/F227L/S287G/A328W/Y332G the mutant has a 1130fold improved catalytic efficiency against (-)-cocaine compared to the wild type enzyme 714226 3.1.1.84 A199S/F227V/S287G/A328W/Y332G the mutant has a 1490fold improved catalytic efficiency against (-)-cocaine compared to the wild type enzyme 714226 3.1.1.84 A199S/S287G/A328W/Y332G the mutant has a 1080fold improved catalytic efficiency against (-)-cocaine compared to the wild type enzyme 714226 3.1.1.84 A199S/S287G/A328W/Y332G the mutant shows 1080fold improved (-)-cocaine-hydrolyzing activity compared to the wild type enzyme 716278 3.1.1.84 A199S/S287G/A328W/Y332G the mutant shows high activity towards (-)-cocaine with about 1080fold (un-fused) and 100fold (when fused with human serum albumin) improved catalytic efficiency compared to the wild type enzyme and also leads to a decrease in catalytic efficiency with acetylthiocholine and butyrylthiocholine 714702 3.1.1.84 A199S/S287G/A328W/Y332G/L286I the mutant has a 242fold improved catalytic efficiency against (-)-cocaine compared to the wild type enzyme 714226 3.1.1.84 A328W/Y332A the mutant has a 9.4fold improved catalytic efficiency against (-)-cocaine compared to the wild type enzyme 714226 3.1.1.84 A328W/Y332G the mutant has a 15fold improved catalytic efficiency against (-)-cocaine compared to the wild type enzyme 714226 3.1.1.84 D259N mutation results in more than 1500fold decrease in kcat 650108 3.1.1.84 F227A/S287G/A328W/Y332M the mutant has a 34fold improved catalytic efficiency against (-)-cocaine compared to the wild type enzyme 714226 3.1.1.84 F261A mutant catalyzed the hydrolysis of cocaine with a 29fold lower kcat and 15fold higher KM 650108 3.1.1.84 F408A mutant has 8fold increased KM and more than 100fold decrease in kcat 650108 3.1.1.84 G173Q kcat and Km-value for cocaine is similar to wild-type value, half-life is increased 7fold compared to wild-type enzyme 694267 3.1.1.84 G173Q the mutant does not have any deleterious effects on the catalytic efficiency -, 716824 3.1.1.84 G173Q/L169K the mutant has a half-life of 370 min and 2.9 days at 37°C 716281 3.1.1.84 G4C/S10C the mutant shows about 4fold reduced catalytic efficiency compared to the wild type enzyme. The mutant retains almost all activity after 7 days of 37°C treatment 716281 3.1.1.84 H287A mutation results in more than 1500fold decrease in kcat 650108 3.1.1.84 L169K the mutation significantly increases the stability of cocaine esterase over that of wild type enzyme (half-life at 37°C is 570 min). The mutant exhibits about 8fold increase in Km for cocaine compared to the wild type enzyme -, 716824 3.1.1.84 L169K/G173Q highly thermostable mutant with a half-life of 2.9 days at 37°C -, 716276 3.1.1.84 L407A mutant has 2fold increased KM and more than 100fold decrease in kcat 650108 3.1.1.84 L407A/F408A attempts to express the L407A/F408A double mutant do not result in any soluble protein 650108 3.1.1.84 LMWP-S-S-T172R/G173Q the mutant shows reduced catalytic efficiency compared to the wild type enzyme -, 730421 3.1.1.84 LMWP-T172R/G173Q the mutant shows reduced catalytic efficiency compared to the wild type enzyme -, 730421 3.1.1.84 additional information computational-experimental effort yields a CocE variant with a 30-fold increase in plasma half-life both in vitro and in vivo 694267 3.1.1.84 Q55E the mutation within the active site of cocE results in a 2fold improvement in KM, but a 14fold loss of kcat 650108 3.1.1.84 S117A mutation results in more than 1500fold decrease in kcat, crystal structures of the S117A and Y44F mutants of cocE. The first urea unfolding transition in the S117A mutant is shifted from 0.5 to 1.3 M urea compared to the wild-type, while the second transition, although broader, has a similar transition point 650108 3.1.1.84 T172R kcat and Km-value for cocaine is similar to wild-type value, half-life is increased 7fold compared to wild-type enzyme 694267 3.1.1.84 T172R the mutants shows about wild type thermal stability and decreased catalytic efficiency for cocaine -, 716824 3.1.1.84 T172R/G173Q kcat and Km-value for cocaine is similar to wild-type value, half-life is increased 30fold compared to wild-type enzyme 694267 3.1.1.84 T172R/G173Q mutant enzyme with increased half-life 699760 3.1.1.84 T172R/G173Q the mutant has an improved in vitro half-life of about 6 h at 37°C -, 728856 3.1.1.84 T172R/G173Q the mutant shows about 4fold reduced catalytic efficiency compared to the wild type enzyme. The mutant remains more than 90% active for longer than 40 days at 37°C, representing a more than 4700fold improvement over wild type. PEGylated mutant enzyme retains full in vitro enzymatic activity 716281 3.1.1.84 T172R/G173Q the mutation extends half-life at 37°C up to 370 min (30fold improvement compared to the wild type stability) and leads to about 3fold decrease of catalytic efficiency for cocaine -, 716824 3.1.1.84 T172R/G173Q -LMWP the mutant shows reduced catalytic efficiency compared to the wild type enzyme -, 730421 3.1.1.84 T172R/G173Q -YGRKKRRQRRR the mutant shows reduced catalytic efficiency compared to the wild type enzyme -, 730421 3.1.1.84 T172R/G173Q /G4C/S10C the mutant shows improved catalytic efficiency against cocaine by about 20% -, 728856 3.1.1.84 T172R/G173Q/L169K the mutant shows poor enzyme kinetics and does not display enhanced stabilization -, 716824 3.1.1.84 T172R/G173Q/L196C/I301C the mutant has not only considerably extended the in vitro half-life at 37°C to more than 100 days, but also significantly improved catalytic efficiency against cocaine by about 150% -, 728856 3.1.1.84 W151A mutant catalyzed the hydrolysis of cocaine with a 78fold lower kcat and 80fold higher KM 650108 3.1.1.84 W166A mutant has a 29fold lower kcat, and a 6fold increased KM 650108 3.1.1.84 Y44F mutation results in more than 1500fold decrease in kcat, crystal structures of the S117A and Y44F mutants of cocE. The urea unfolding curve of the Y44F mutant is very similar to the wild-type, and has almost identical transition points 650108 3.1.1.84 YGRKKRRQRRR-T172R/G173Q the mutant still maintains 52% of its cocaine-hydrolyzing efficiency even after incubation at 37°C for 24 h -, 730421