Sequence- and activity-based screening of microbial genomes for novel dehalogenases
Article first published online: 12 NOV 2009
© 2009 The Authors. Journal compilation © 2009 Society for Applied Microbiology and Blackwell Publishing Ltd
Special Issue: Biocatalysis. Guest Editors: Karl-Erich Jaeger, Andreas Schmid and Manuel Ferrer
Volume 3, Issue 1, pages 107–120, January 2010
How to Cite
Chan, W. Y., Wong, M., Guthrie, J., Savchenko, A. V., Yakunin, A. F., Pai, E. F. and Edwards, E. A. (2010), Sequence- and activity-based screening of microbial genomes for novel dehalogenases. Microbial Biotechnology, 3: 107–120. doi: 10.1111/j.1751-7915.2009.00155.x
- Issue published online: 20 DEC 2009
- Article first published online: 12 NOV 2009
- Received 24 August, 2009; revised 21 September, 2009; accepted 24 September, 2009.
Fig. S1. Representative SDS-PAGE analysis of P. aeruginosa protein targets.
The target proteins in well-expressed and soluble samples appear as large round patches on the gel (e.g. lanes C8, C10, A8 and C6). By inspection, the purity of these high-yield samples is generally above 75%. For samples that are poorly expressed or insoluble, a large number of expression host protein bands are detected. In such cases (e.g. lanes E1, E2, E5 and E7), the protein purity is approximately 35% at best. The identifiable target protein bands in low-yield samples have been circled. MW denotes molecular weight of the standards in kDa.
Fig. S2. Representative data in carboxylesterase and thioesterase screens.
The upper and lower panels display representative data from the carboxylesterase and thioesterase screens respectively. Absorbance was measured 3 h (light bars) and 24 h (dark bars) after initiation of the reaction. Data from the 24-hour time point for the carboxylesterase screen was not interpretable due to high backgrounds. The dashed lines mark the cut-off used for confirming activities.
Table S1. All ABH superfamily targets selected for biochemical characterization.
Table S2. All HAD superfamily targets selected for biochemical characterization.
Table S3. Cloning and purification results.
Table S4. Summary of cloning and purification results.
Table S5. Sequence identity and similarity of all predicted haloalkane dehalogenases.
Table S6. Sequence identity and similarity of all predicted fluoroacetate dehalogenases.
Table S7. Sequence identity and similarity of all predicted l-2-haloacid dehalogenases.
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Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.