Enzymes with {high|higher} activity are readily {produced|created|made|developed} {through|via|by means of|by way of} protein engineering, but intentionally and {efficiently|effectively} engineering enzymes for an expanded scope {is a|is really a|is actually a|can be a|is often a|is usually a} {contemporary|modern} challenge. Measuring reaction outcomes on mixtures of substrates, {called|known as|referred to as|named} {here|right here} SUbstrate Multiplexed Screening (SUMS), has {long|lengthy|extended} been {used|utilized|employed|utilised|applied|made use of} to rigorously quantitate enzyme specificity. {Despite|Regardless of|In spite of} the {potential|possible|prospective} utility of SUMS to guide engineering of promiscuous enzymes, this {approach|method|strategy} has not {found|discovered|identified|located} widespread adoption in biocatalysis. {Here|Right here}, we {develop|create} principles of {how to|how you can|the best way to|the way to|tips on how to|ways to} {design|style|design and style} robust SUMS {methods|techniques|strategies|approaches|procedures|solutions} that, {rather than|instead of|as opposed to|as an alternative to|in lieu of} assess absolute specificity, use heuristic readouts of substrate promiscuity to {identify|determine|recognize} hits for {further|additional} investigation. This {rich|wealthy} {information|info|details|data|facts|information and facts} enables engineering of activity for {multiple|numerous|several|a number of|many|various} substrates simultaneously and identifies enzyme variants with altered promiscuity, even when {overall|general|all round} activity is {lower|reduce|reduced|decrease}. We demonstrate the effectiveness of SUMS by engineering two enzymes to {produce|create|generate|make} pharmacologically active tryptamines from {simple|easy|straightforward|basic|uncomplicated|very simple} indole precursors {in a|inside a|within a} biocatalytic cascade. These advances leverage {common|typical|frequent|widespread|prevalent|popular} laboratory {equipment|gear} and represent a {highly|extremely|very|hugely} accessible and customizable {method|technique|approach|strategy|system|process} for enzyme engineering. Ethyl 2-chloropyrimidine-5-carboxylate Price 1471260-52-2 Chemscene PMID:23255394
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