Chemoenzymatic elaboration of monosaccharides using engineered cytochrome P450 BM3 demethylases

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 106; no. 39; pp. 16550 - 16555
• Main Authors: Lewis, Jared C., Bastian, Sabine, Bennett, Clay S., Fu, Yu, Mitsuda, Yuuichi, Chen, Mike M., Greenberg, William A., Wong, Chi-Huey, Arnold, Frances H.
• Format: Journal Article
• Language: English
• Published: 29.09.2009
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.0908954106

Polysaccharides comprise an extremely important class of biopolymers that play critical roles in a wide range of biological processes, but the synthesis of these compounds is challenging because of their complex structures. We have developed a chemoenzymatic method for regioselective deprotection of monosaccharide substrates using engineered Bacillus megaterium cytochrome P450 (P450 BM3 ) demethylases that provides a highly efficient means to access valuable intermediates, which can be converted to a wide range of substituted monosaccharides and polysaccharides. Demethylases displaying high levels of regioselectivity toward a number of protected monosaccharides were identified using a combination of protein and substrate engineering, suggesting that this approach ultimately could be used in the synthesis of a wide range of substituted mono- and polysaccharides for studies in chemistry, biology, and medicine.