Title : Unscrambling thermal stability and temperature adaptation in evolved variants of a cold-active lipase - Gatti-Lafranconi_2008_FEBS.Lett_582_2313 |
Author(s) : Gatti-Lafranconi P , Caldarazzo SM , Villa A , Alberghina L , Lotti Marina , Lotti M |
Ref : FEBS Letters , 582 :2313 , 2008 |
Abstract :
Directed evolution by error-prone PCR was applied to stabilize the cold-active lipase from Pseudomonas fragi (PFL). PFL displays high activity at 10 degrees C, but it is highly unstable even at moderate temperatures. After two rounds of evolution, a variant was generated with a 5-fold increase in half-life at 42 degrees C and a shift of 10 degrees C in the temperature optimum, nevertheless retaining cold-activity. The evolved lipase displayed specific activity higher than the wild type enzyme in the temperature range 29-42 degrees C. Biophysical measurements did not indicate any obvious difference between the improved variant and the wild type enzyme in terms of loss of secondary structure upon heat treatment, nor a shift in the apparent melting temperature. |
PubMedSearch : Gatti-Lafranconi_2008_FEBS.Lett_582_2313 |
PubMedID: 18534193 |
Gatti-Lafranconi P, Caldarazzo SM, Villa A, Alberghina L, Lotti Marina, Lotti M (2008)
Unscrambling thermal stability and temperature adaptation in evolved variants of a cold-active lipase
FEBS Letters
582 :2313
Gatti-Lafranconi P, Caldarazzo SM, Villa A, Alberghina L, Lotti Marina, Lotti M (2008)
FEBS Letters
582 :2313