In:
Science, American Association for the Advancement of Science (AAAS), Vol. 311, No. 5760 ( 2006-01-27), p. 535-538
Abstract:
The design of enzymes with new functions and properties has long been a goal in protein engineering. Here, we report a strategy to change the catalytic activity of an existing protein scaffold. This was achieved by simultaneous incorporation and adjustment of functional elements through insertion, deletion, and substitution of several active site loops, followed by point mutations to fine-tune the activity. Using this approach, we were able to introduce β-lactamase activity into the αβ/βα metallohydrolase scaffold of glyoxalase II. The resulting enzyme, evMBL8 (evolved metallo β-lactamase 8), completely lost its original activity and, instead, catalyzed the hydrolysis of cefotaxime with a ( k cat / K m ) app of 1.8 × 10 2 (mole/liter) –1 second –1 , thus increasing resistance to Escherichia coli growth on cefotaxime by a factor of about 100.
Type of Medium:
Online Resource
ISSN:
0036-8075
,
1095-9203
DOI:
10.1126/science.1118953
Language:
English
Publisher:
American Association for the Advancement of Science (AAAS)
Publication Date:
2006
detail.hit.zdb_id:
128410-1
detail.hit.zdb_id:
2066996-3
detail.hit.zdb_id:
2060783-0
SSG:
11
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