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The specific epoxidation activity of the most active mutant S96A toward styrene and trans-β-methylstyrene are 2.6- and 2.3-fold of the wild-type, respectively.
The specific epoxidation activity of the most active mutant S96A toward styrene and trans-β-methyl styrene were 2.6 and 2.3-fold of the wild-type, respectively.
The most active mutant T210G was further mutated at position 116, but no mutant showed enhanced catalytic activity.
Other heme peroxidases were less active by orders of magnitude; protein engineering has resulted in impressive improvements but even the most active mutant was still at least an order of magnitude less active than CPO.
A complete screen of the single-substitution GFP mutants was carried out (except for F83/F84 where only 202 cfu were required to find the most active mutant among all the single-substitution variants).
EGFR/D770insNPG, the most active mutant, was insensitive to gefitinib (0.1 μ M, 5 h), as were the less active mutants EGFR/D770InsNPH and EGFR/M766InsASV.
Similar(52)
The two most active mutants showed up to 70-fold higher catalytic efficiency than the parental GST A2-2.
Interestingly, the phenylalanine-mutants, designed as negative controls were the most active mutants which suggests rather a structural role of His106.
Then, plasmid DNA for the 19 most active mutants was extracted and retransformed into naïve yeast for detailed analysis.
The most active mutants were selected, characterised and used as a template in subsequent rounds of ep-PCR.
The most active mutants were designated as PI5P4Kβ+ (PI5P4Kβ G1, Figure 3) and PI5P4Kγ+ (PI5P4Kγ G3+AB, Figure 3) and enzyme turnover rates were quantified and calculated for these mutants for comparison with the wild-type enzymes (Table 1 and Supplementary Figure S1).
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