Sterol 3β-glucosyltransferase biocatalysts with a range of selectivities, including selectivity for testosterone.
The main objectives of this work were to characterise a range of purified recombinant sterol 3β-glucosyltransferases and show that rational sampling of the diversity that exists within sterol 3β-glucosyltransferase sequence space can result in a range of enzyme selectivities. In our study the catalytically active domain of the Saccharomyces cerevisiae 3β-glucosyltransferase was used to mine putative sterol 3β-glucosyltransferases from the databases. Selected diverse sequences were expressed in and purified from Escherichia coli and shown to have different selectivities for the 3β-hydroxysteroids ergosterol and cholesterol. Surprisingly, three enzymes were also selective for testosterone, a 17β-hydroxysteroid. This study therefore reports for the first time sterol 3β-glucosyltransferases with selectivity for both 3β- and 17β-hydroxysteroids and is also the first report of recombinant 3β-glucosyltransferases with selectivity for steroids with a hydroxyl group at positions other than C-3. These enzymes could therefore find utility in the pharmaceutical industry for the green synthesis of a range of glycosylated compounds of medicinal interest.
Industrial Biotechnology Research Group, Department of Applied Sciences, Faculty of Health and Life Sciences, Northumbria University, Newcastle upon Tyne, NE1 8ST, UK. gary.black@northumbria.ac.uk.
Vatsala
V
Malik
Meng
M
Zhang
Lynn G
LG
Dover
Julian S
JS
Northen
Anthony
A
Flinn
Justin J
JJ
Perry
Gary W
GW
Black
D000595:Amino Acid Sequence; D002384:Catalysis; D020134:Catalytic Domain; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005964:Glucosyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D054730:Protein Interaction Domains and Motifs; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013261:Sterols / Q000378:metabolism*; D013379:Substrate Specificity; D013739:Testosterone / Q000378:metabolism*
D013261:Sterols; D013739:Testosterone; D005964:Glucosyltransferases