this is for holding javascript data
Alex Carlin edited Discussion__.md
almost 8 years ago
Commit id: 537294bfd8f67e85001042fd7dc1c8d7fb9a01b1
deletions | additions
diff --git a/Discussion__.md b/Discussion__.md
index e9e58ec..b1c4330 100644
--- a/Discussion__.md
+++ b/Discussion__.md
...
# Discussion
[...] The introduction of catalytic activity into existing enzyme scaffolds has been used for all successful enzyme designs to date. The choice of a suitable protein "scaffold" on which to design mutations is of paramount importance in the sucessful design of enzymes de novo. The redesign of existing catalytic proteins for new substrates is also hughly dependent on the inital starting point: the scaffold protein of a particular fold that is used to design mutations.
The traditional dogma in the field of enzyme engineering is that thermostable proteins make the best scaffolds. They are said to possess intrinsic resistence to destabilizing effects of mutations. However, very few proteins have been systemically characterized for their ability to retain stability and activity when mutations are made across large portions of the protein. Thus, efforts to add data to the assertion that one should begin design efforts with a thermostable protein are necessary. Previous work provides a mounting body of evidence that the claim that protein design should process from thermostable scaffolds is mounting. The work of Brian Matthews on hundreds of point mutants of T7 lysozyme is a prime example: almost all of the point mutants retained their fold and were crystalized. Conversely, a 1998 paper on the stability of proteins at temperature above 100 C introduced a double mutation that increased thermostability about 8 fold.
In this study, we functionally characterized over 100 single point mutants of β-glucosidase B from P. polymxa at various temperatures ranging from 30 C to 50 C via a colorimetric assay. We found that, contrary to the accepted dogma, point mutants of this enzyme did not differ in Tm from the wild type enzyme by more than X ± Y C. Out of 83 mutants assayed, we found a mean Tm of 40 ± 0.7 C within the range 37.6–41.4 C.
