کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2818208 | 1160035 | 2012 | 8 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Engineering of a metagenome derived lipase toward thermal tolerance: Effect of asparagine to lysine mutation on the protein surface Engineering of a metagenome derived lipase toward thermal tolerance: Effect of asparagine to lysine mutation on the protein surface](/preview/png/2818208.png)
A highly thermostable mutant lipase was generated and characterized. Mutant enzyme demonstrated 144 fold enhanced thermostability over the wild type enzyme at 60 °C. Interestingly, the overall catalytic efficiency (kcat/Km) of mutant was also enhanced (~ 20 folds). Circular dichroism spectroscopy, studied as function of temperature, demonstrated that the mutant lipase retained its secondary structure up to 70–80 °C, whereas wild type protein structure was completely distorted above 35 °C. Additionally, the intrinsic tryptophan fluorescence (a probe for the tertiary structure) also displayed difference in the conformation of two enzymes during temperature dependent unfolding. Furthermore, mutation N355K resulted in extensive H-bonding (Lys355 HZ1OE2 Glu284) with a distance 2.44 Å. In contrast to this, Wt enzyme has not shown such H-bonding interaction.
► This study reported characterization of highly thermostable mutant lipase.
► Mutation resulted in substitution of amino acid N355K in mature polypeptide.
► Mutation improved both thermal stability and catalytic efficiency of the lipase protein.
► Mutant protein retained its secondary structure at high temperature compared to Wt protein.
► Mutant N355K resulted in extensive H bonding, in contrast to Wt enzyme.
Journal: Gene - Volume 491, Issue 2, 10 January 2012, Pages 264–271