High GC content gene synthesis
This success story was provided by the group of Prof. Dr. Bechthold (Pharmaceutical Biology, Albert-Ludwigs-University, Freiburg)
This synthetic gene was prepared by Mr Gene while other companies refused to synthesize the gene due to a very high GC content.
Order submitted: March 18th - gene finalized: April 17th
Length: 471 bp - GC content: 71 %
The pharmaceutical activity of many natural products, among them valuable antibiotics and anticancer therapeutics, depends on regio- and stereospecifically attached sugar moieties. The attachment of these sugars is catalyzed by glycosyltransferases (GTs). The acceptor substrate of glycosyltransferases vary widely, the donor substrate is almost always an activated sugars, with the most common activated species being NDP sugars. Landomycin A and landomycin E, both produced by different Streptomyces strains, belong to the angucycline type of natural products and possesses strong antitumor activities, in particular against prostate cancer cell lines. Detailed studies on the biosynthesis of the hexasaccharide side chain of landomycin A revealed that four glycosyltransferases are responsible for the formation of the side chain. LanGT2 is the priming glycosyltransferase connecting D-olivose to the landomycin aglycon (landomycinone) and LanGT3 is the olivosyltransferase responsible for the transfer of the fourth sugar moiety. LanGT1 is an iteratively acting olivosyltransferase catalyzing the attachment of the second and the fifth sugar and LanGT4 is an iteratively acting rhodinosyltransferase attaching the third and sixth sugar during landomycin A biosynthesis. Three glycosyltransferases are responsible for the formation of landomycin E in S. globisporus. Here LndGT2 is the priming glycosyltransferase, LndGT1 catalyzes the attachment of the second sugar and LndGT4 attaches the third sugar. We have shown that LndGT4 can not act twice during landomycin biosynthesis. The aim of studies was to detect amino acids responsible for the iterative work of LanGT4 and introduce them into LndGT4. We were able to design a gene encoding an iteratively working enzyme and Mr. Gene did the synthesis.
Thank you Mr. Gene!
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