Chemoenzymatic elaboration of monosaccharides using engineered cytochrome P450BM3 demethylases.
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.
Lewis
Jared C
JC
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA 91125, USA.
Bastian
Sabine
S
Bennett
Clay S
CS
Fu
Yu
Y
Mitsuda
Yuuichi
Y
Chen
Mike M
MM
Greenberg
William A
WA
Wong
Chi-Huey
CH
Arnold
Frances H
FH
D001410:Bacillus megaterium / Q000201:enzymology* / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002240:Carbohydrate Sequence; D002384:Catalysis; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D009005:Monosaccharides / Q000737:chemistry / Q000378:metabolism*; D010089:Oxidoreductases, N-Demethylating / Q000737:chemistry* / Q000378:metabolism; D011487:Protein Conformation; D015202:Protein Engineering; D013379:Substrate Specificity
D001426:Bacterial Proteins; D009005:Monosaccharides; D003577:Cytochrome P-450 Enzyme System; D010089:Oxidoreductases, N-Demethylating