The kinetic model of the shikimate pathway as a tool to optimize enzyme assays for high-throughput screening
Four‐enzyme section of the shikimate pathway (Aro B, D, E, and K) of Streptococcus pneumoniae has been studied. Kinetic properties of the individual enzymes and three‐ and four‐enzyme linked reactions have been characterized in vitro. On the basis of the data measured in spectrophotometric and LC‐MS...
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Published in | Biotechnology and bioengineering Vol. 95; no. 4; pp. 560 - 571 |
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Main Authors | , , , , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
Hoboken
Wiley Subscription Services, Inc., A Wiley Company
05.11.2006
Wiley Subscription Services, Inc |
Subjects | |
Online Access | Get full text |
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Summary: | Four‐enzyme section of the shikimate pathway (Aro B, D, E, and K) of Streptococcus pneumoniae has been studied. Kinetic properties of the individual enzymes and three‐ and four‐enzyme linked reactions have been characterized in vitro. On the basis of the data measured in spectrophotometric and LC‐MS experiments, kinetic mechanisms of the enzymes have been suggested and all kinetic parameters have been identified. Kinetic models for these three‐ and four‐enzyme sections of the shikimate pathway have been constructed and validated. The model of the four‐enzyme section of shikimate pathway has been employed to design an inhibition‐sensitive reconstituted pathway for a high‐throughput screening effort on the shikimate pathway. It was demonstrated that using the model it was possible to optimize this reconstituted pathway in such a way to provide equal sensitivity of the enzymes to inhibition. © 2006 Wiley Periodicals, Inc. |
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Bibliography: | ark:/67375/WNG-HV38D5CN-P ArticleID:BIT20772 istex:C4ACB217F03281A1DA0B14B6530855675B1BEF88 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 0006-3592 1097-0290 |
DOI: | 10.1002/bit.20772 |