|
type |
Journal Article |
authors |
Aron ZD, Dorrestein PC, Blackhall JR, Kelleher NL, Walsh CT. |
title |
Characterization of a new tailoring domain in polyketide biogenesis: the amine transferase domain of MycA in the mycosubtilin gene cluster. |
journal |
J Am Chem Soc. |
Activity |
pks-atd |
Family |
pks-atd |
sel |
selected |
ui |
16248612 |
year |
(2005) |
volume |
127 |
number |
43 |
pages |
14986-7 |
| |
abstract |
We report the expression and characterization of a truncated form of MycA from the Mycosubtilin gene cluster from Bacillus subtilis. The MycA fragment contains a new amino transferase (AMT) tailoring domain, allowing the first detailed study of a PLP-dependent enzyme operating in cis within the PKS and NRPS biosynthetic paradigm. As the AMT domain acts on covalently bound beta-ketothioesters, and is therefore a single-turnover system, electrospray ionization-Fourier transform mass spectrometry (ESI-FTMS) was used to observe the amine-transfer reaction both for amine donor substrate specificity and to regiospecifically determine enzyme-bound intermediates. We confirm the function of the AMT domain, dissect the mechanistic steps of amine transfer, identify the preferred amine source, and localize the beta-ketothioester substrate during amine transfer. |
last changed |
2018/03/27 09:55 |
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