Directed evolution of bright mutants of an oxygen-independent flavin-binding fluorescent protein from Pseudomonas putida

Table 2 Classification of mutants

Mutation	Beneficial	Deleterious	Neutral
F37X	2	71	103
D52X	0	42	138
A53X	0	60	57
R54X	0	28	152
Q57X	0	30	150
R70X	0	18	120
N85X	0	141	20
W94X	0	15	58
Y112X	0	31	149

Mutations were introduced in the wild type FbFP by site saturation mutagenesis and mutants were classified as beneficial, neutral, or deleterious based on their fluorescent properties. Specifically, beneficial mutants exhibit at least a twofold enhancement in brightness of fluorescence emission (or a 10 nm or greater spectral shift), deleterious mutants have substantially diminished (greater than twofold) fluorescence emission compared to the wild type FbFP, and neutral mutants are similar to the wild type proteins with respect to brightness and emission wavelength.

ISSN: 1754-1611