RebG- and RebM-catalyzed indolocarbazole diversification.
Rebeccamycin and staurosporine represent two broad classes of indolocarbazole glycoside natural products with antitumor properties. Based upon previous sequence annotation and in vivo studies, rebG encodes for the rebeccamycin N-glucosyltransferase, and rebM for the requisite 4'-O-methyltransferase. In the current study, an efficient in vivo biotransformation system for RebG was established in both Streptomyces lividans and Escherichia coli. Bioconversion experiments revealed RebG to glucosylate a set of indolocarbazole surrogates, the products of which could be further modified by in vitro RebM-catalyzed 4'-O-methylation. Both RebG and RebM displayed substrate promiscuity, and evidence for a remarkable lack of RebG regioselectivity in the presence of asymmetric substrates is also provided. In the context of the created indolocarbazole analogues, cytotoxicity assays also highlight the importance of 4'-O-methylation for their biological activity.
Laboratory for Biosynthetic Chemistry, Pharmaceutical Sciences Division, School of Pharmacy, 777 Highland Avenue, Madison, WI 53705, USA.
Changsheng
C
Zhang
Christoph
C
Albermann
Xun
X
Fu
Noel R
NR
Peters
John D
JD
Chisholm
Guisheng
G
Zhang
Eric J
EJ
Gilbert
Peng George
PG
Wang
David L
DL
Van Vranken
Jon S
JS
Thorson
D001426:Bacterial Proteins / Q000737:chemistry*; D002227:Carbazoles / Q000138:chemical synthesis* / Q000737:chemistry; D002384:Catalysis; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D005964:Glucosyltransferases / Q000737:chemistry*; D006031:Glycosylation; D006801:Humans; D066298:In Vitro Techniques; D007211:Indoles / Q000737:chemistry*; D008780:Methyltransferases / Q000737:chemistry*; D015394:Molecular Structure; D013997:Time Factors; D014407:Tumor Cells, Cultured
D001426:Bacterial Proteins; D002227:Carbazoles; D007211:Indoles; D008780:Methyltransferases; D005964:Glucosyltransferases