TY - JOUR
T1 - Identification of genes essential for fluorination and sulfamylation within the nucleocidin gene clusters of Streptomyces calvus and Streptomyces virens
AU - Wojnowska, Marta
AU - Feng, Xuan
AU - Chen, Yawen
AU - Deng, Hai
AU - O'Hagan, David
N1 - Acknowledgements
We thank the EPSRC, Leverhulme Trust and the EU Horizon 2020 Sinfonia consortia for financial support. The Chinese Scholarship Council is acknowledged for studentship support (Y.C.). We also thank Pfizer Ltd., (Groton, USA) for supplying the nucleocidin-producing strain S. calvus T-3018.
Research Funding
Leverhulme Trust. Grant Number: RPG-2021-028
Chinese Scholarship Council
Engineering and Physical Sciences Research Council
PY - 2023/3/1
Y1 - 2023/3/1
N2 - The gene cluster in Streptomyces calvus associated with the biosynthesis of the fluoro- and sulfamyl-metabolite nucleocidin was interrogated by systematic gene knockouts. Out of the 26 gene deletions most did not affect fluorometabolite production, nine abolished sulfamylation but not fluorination, and three precluded fluorination, having no effect on sulfamylation. In addition to nucI, nucG, nucJ, nucK, nucL, nucN, nucO, nucQ and nucP we identified two genes (nucW, nucA), belonging to a phosphoadenosine phosphosulfate (PAPS) gene cluster, as required for sulfamyl assembly. Three genes (orf(-3), orf2 and orf3) were found to be essential for fluorination, although the activities of their protein products are unknown. These genes as well as nucK, nucN, nucO and nucPNP, whose knockouts produced results differing from those described in a recent report, were also deleted in Streptomyces virens - with confirmatory outcomes. This genetic profile should inform biochemistry aimed at uncovering the enzymology behind nucleocidin biosynthesis.
AB - The gene cluster in Streptomyces calvus associated with the biosynthesis of the fluoro- and sulfamyl-metabolite nucleocidin was interrogated by systematic gene knockouts. Out of the 26 gene deletions most did not affect fluorometabolite production, nine abolished sulfamylation but not fluorination, and three precluded fluorination, having no effect on sulfamylation. In addition to nucI, nucG, nucJ, nucK, nucL, nucN, nucO, nucQ and nucP we identified two genes (nucW, nucA), belonging to a phosphoadenosine phosphosulfate (PAPS) gene cluster, as required for sulfamyl assembly. Three genes (orf(-3), orf2 and orf3) were found to be essential for fluorination, although the activities of their protein products are unknown. These genes as well as nucK, nucN, nucO and nucPNP, whose knockouts produced results differing from those described in a recent report, were also deleted in Streptomyces virens - with confirmatory outcomes. This genetic profile should inform biochemistry aimed at uncovering the enzymology behind nucleocidin biosynthesis.
KW - nucleocidin
KW - fluorometabolite biosynthesis
KW - Streptomyces
KW - gene knockouts
KW - natural products
U2 - 10.1002/cbic.202200684
DO - 10.1002/cbic.202200684
M3 - Article
C2 - 36548247
VL - 24
JO - ChemBioChem
JF - ChemBioChem
SN - 1439-4227
IS - 5
M1 - e202200684
ER -