Structurally informed site-directed mutagenesis of a stereochemically promiscuous aldolase to afford stereochemically complementary biocatalysts

Sylvain F Royer, L Haslett, Susan J Crennell, David W Hough, Michael J Danson, Steven D Bull

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

2-Keto-3-deoxygluconate aldolase from the hyperthermophile Sulfolobus solfataricus is a highly thermostable type I aldolase that can catalyze carbon-carbon bond formation using nonphosphorylated substrates. However, it exhibits poor diastereocontrol in many of its aldol reactions, including the reaction of its natural substrates, pyruvate and D-glyceraldehyde, which afford a 55:45 mixture of D-2-keto-3-deoxygluconate (D-KDGlu) and D-2-keto-3-deoxy-galactonate (D-KDGal). We have employed detailed X-ray crystallographic structural information of this aldolase bound to these diastereoisomeric aldol products to selectively target specific amino acids for mutation for the rapid creation of stereochemically complementary mutants that catalyze either (Re)- or (Si)-facial selective aldol reactions to afford either D-KDGlu or D-KDGal with good levels of diastereocontrol.
Original languageEnglish
Pages (from-to)11753-11758
Number of pages6
JournalJournal of the American Chemical Society
Volume132
Issue number33
Early online date3 Aug 2010
DOIs
Publication statusPublished - 25 Aug 2010

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