Calicheamicin gamma1 from Micromonospora echinospora spp. calichensis is the most prominent of the 10- membered enediyne family with respect to its unprecedented molecular architecture, spectacular biological activity and clinical value. As such, calicheamicin is an excellent target for the study of natural product biosynthesis and self-resistance. The objective of the first phase of this study was to i) pursue the biosynthesis of the DNA-delivery component of calicheamicin (the aryltetrasaccharide, comprised of four uniquely functionalized sugars), ii) develop the genetic tools (transformation and gene disruption protocols) to address calicheamicin biosynthesis in Micromonospora and iii) investigate the mechanism(s) of calicheamicin self-resistance in Micromonospora. With these goals achieved and new tools/information in place, the second phase of this massive project will predominately focus upon expanding this program toward understanding and exploiting the complex biosynthesis of the enediyne core. While continuing our focus upon calicheamicin as a model for 10-membered enediyne biosynthesis, a second complimentary 10-membered enediyne model will be pursued (namely, dynemicin from Micromonospora chersina) selected for its unique architecture (an unprecedented fused enediyne-anthracycline), predominate biological activity, anticipated small gene locus size (excellent for production of dynemicin in 'genetically-friendly' heterologous hosts) and the opportunity for comparative genomics of the calicheamicin and dynemicin biosynthetic loci. The fundamental vision of this program remains constant - to present rational strategies from which to build a foundation of knowledge regarding 10-membered enediyne biosynthesis and self-resistance; the consequence of which will continue to provide pioneering discoveries in enzymatic transformation, tools for the rational biosynthetic modification of natural product drug leads, the potential for enediyne overproducing strains and possibly even an enediyne combinatorial biosynthesis program.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA084374-10
Application #
7432499
Study Section
Bio-Organic and Natural Products Chemistry Study Section (BNP)
Program Officer
Fu, Yali
Project Start
1999-12-01
Project End
2010-05-31
Budget Start
2008-06-01
Budget End
2010-05-31
Support Year
10
Fiscal Year
2008
Total Cost
$275,409
Indirect Cost
Name
University of Wisconsin Madison
Department
Type
Schools of Pharmacy
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
Cao, Hongnan; Tan, Kemin; Wang, Fengbin et al. (2016) Structural dynamics of a methionine ?-lyase for calicheamicin biosynthesis: Rotation of the conserved tyrosine stacking with pyridoxal phosphate. Struct Dyn 3:034702
Elshahawi, Sherif I; Shaaban, Khaled A; Kharel, Madan K et al. (2015) A comprehensive review of glycosylated bacterial natural products. Chem Soc Rev 44:7591-697
Singh, Shanteri; Michalska, Karolina; Bigelow, Lance et al. (2015) Structural Characterization of CalS8, a TDP-?-D-Glucose Dehydrogenase Involved in Calicheamicin Aminodideoxypentose Biosynthesis. J Biol Chem 290:26249-58
Singh, Shanteri; Kim, Youngchang; Wang, Fengbin et al. (2015) Structural characterization of AtmS13, a putative sugar aminotransferase involved in indolocarbazole AT2433 aminopentose biosynthesis. Proteins 83:1547-54
Peltier-Pain, Pauline; Singh, Shanteri; Thorson, Jon S (2015) Characterization of Early Enzymes Involved in TDP-Aminodideoxypentose Biosynthesis en Route to Indolocarbazole AT2433. Chembiochem 16:2141-6
Wang, Fengbin; Singh, Shanteri; Xu, Weijun et al. (2015) Structural Basis for the Stereochemical Control of Amine Installation in Nucleotide Sugar Aminotransferases. ACS Chem Biol 10:2048-56
Wang, Fengbin; Singh, Shanteri; Zhang, Jianjun et al. (2014) Understanding molecular recognition of promiscuity of thermophilic methionine adenosyltransferase sMAT from SulfolobusĀ solfataricus. FEBS J 281:4224-39
Singh, Shanteri; Zhang, Jianjun; Huber, Tyler D et al. (2014) Facile chemoenzymatic strategies for the synthesis and utilization of S-adenosyl-(L)-methionine analogues. Angew Chem Int Ed Engl 53:3965-9
Elshahawi, Sherif I; Ramelot, Theresa A; Seetharaman, Jayaraman et al. (2014) Structure-guided functional characterization of enediyne self-sacrifice resistance proteins, CalU16 and CalU19. ACS Chem Biol 9:2347-58
Singh, Shanteri; Peltier-Pain, Pauline; Tonelli, Marco et al. (2014) A general NMR-based strategy for the in situ characterization of sugar-nucleotide-dependent biosynthetic pathways. Org Lett 16:3220-3

Showing the most recent 10 out of 59 publications