title: Application of thermophilic enzymes in commercial biotransformation processes.
abstract: Biocatalysis is a useful tool in the provision of chiral technology and extremophilic enzymes are just one component in that toolbox. Their role is not always attributable to their extremophilic properties; as with any biocatalyst certain other criteria should be satisfied. Those requirements for a useful biocatalyst will be discussed including issues of selectivity, volume efficiency, security of supply, technology integration, intellectual property and regulatory compliance. Here we discuss the discovery and commercialization of an L-aminoacylase from Thermococcus litoralis, the product of a LINK project between Chirotech Technology and the University of Exeter. The enzyme was cloned into Escherichia coli to aid production via established mesophilic fermentation protocols. A simple downstream process was then developed to assist in the production of the enzyme as a genetically modified-organism-free reagent. The fermentation and downstream processes are operated at the 500 litre scale. Characterization of the enzyme demonstrated a substrate preference for N-benzoyl groups over N-acetyl groups. The operational parameters have been defined in part by substrate-concentration tolerances and also thermostability. Several examples of commercial biotransformations will be discussed including a process that is successful by virtue of the enzyme's thermotolerance.
authors.affiliation: Dowpharma, Chirotech Technology Ltd, 321 Cambridge Science Park, Milton Road, Cambridge CB4 0WG, U.K. itaylor@dow.com
authors.forename: I N
authors.initials: IN
authors.lastname: Taylor
authors.forename: R C
authors.initials: RC
authors.lastname: Brown
authors.forename: M
authors.initials: M
authors.lastname: Bycroft
authors.forename: G
authors.initials: G
authors.lastname: King
authors.forename: J A
authors.initials: JA
authors.lastname: Littlechild
authors.forename: M C
authors.initials: MC
authors.lastname: Lloyd
authors.forename: C
authors.initials: C
authors.lastname: Praquin
authors.forename: H S
authors.initials: HS
authors.lastname: Toogood
authors.forename: S J C
authors.initials: SJ
authors.lastname: Taylor
mesh_terms: D000581:Amidohydrolases / Q000737:chemistry; D001671:Biochemistry / Q000379:methods*; D019149:Bioreactors; D001709:Biotechnology / Q000379:methods; D001711:Biotransformation; D002384:Catalysis; D004795:Enzyme Stability; D004926:Escherichia coli / Q000378:metabolism; D005285:Fermentation; D006358:Hot Temperature; D008956:Models, Chemical; D013379:Substrate Specificity; D019712:Thermococcus / Q000201:enzymology
chemical_list: D000581:Amidohydrolases; C019672:aminoacylase I