title: Suitability of the hydrocarbon-hydroxylating molybdenum-enzyme ethylbenzene dehydrogenase for industrial chiral alcohol production.
abstract: The molybdenum/iron-sulfur/heme protein ethylbenzene dehydrogenase (EbDH) was successfully applied to catalyze enantiospecific hydroxylation of alkylaromatic and alkylheterocyclic compounds. The optimization of the synthetic procedure involves use of the enzyme in a crude purification state that saves significant preparation effort and is more stable than purified EbDH without exhibiting unwanted side reactions. Moreover, immobilization of the enzyme on a crystalline cellulose support and changes in reaction conditions were introduced in order to increase the amounts of product formed (anaerobic atmosphere, electrochemical electron acceptor recycling or utilization of ferricyanide as alternative electron acceptor in high concentrations). We report here on an extension of effective enzyme activity from 4h to more than 10 days and final product yields of up to 0.4-0.5g/l, which represent a decent starting point for further optimization. Therefore, we expect that the hydrocarbon-hydroxylation capabilities of EbDH may be developed into a new process of industrial production of chiral alcohols.
authors.affiliation: Jerzy Haber Institute of Catalysis and Surface Chemistry, PAS, Niezapominajek 8, 30-239 Kraków, Poland.
authors.forename: M
authors.initials: M
authors.lastname: Tataruch
authors.affiliation: Laboratory for Microbial Biochemistry and Synmikro Center for Synthetic Microbiology, Philipps University of Marburg, 35043 Marburg, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Heider
authors.affiliation: Department of Bioinorganic Chemistry, Wrocław Technical University, Wybrzeże Wyspiańskiego 27, 50-370 Wrocław, Poland.
authors.forename: J
authors.initials: J
authors.lastname: Bryjak
authors.affiliation: Jerzy Haber Institute of Catalysis and Surface Chemistry, PAS, Niezapominajek 8, 30-239 Kraków, Poland.
authors.forename: P
authors.initials: P
authors.lastname: Nowak
authors.affiliation: Jerzy Haber Institute of Catalysis and Surface Chemistry, PAS, Niezapominajek 8, 30-239 Kraków, Poland.
authors.forename: D
authors.initials: D
authors.lastname: Knack
authors.affiliation: Department of Biotechnology and Food Microbiology, Poznan University of Life Sciences, Wojska Polskiego 48, Poznań 60-627, Poland.
authors.forename: A
authors.initials: A
authors.lastname: Czerniak
authors.affiliation: Department of Chemical Technology, Kaunas University of Technology, Radvilenu pl. 19, LT-50254 Kaunas, Lithuania.
authors.forename: J
authors.initials: J
authors.lastname: Liesiene
authors.affiliation: Jerzy Haber Institute of Catalysis and Surface Chemistry, PAS, Niezapominajek 8, 30-239 Kraków, Poland. Electronic address: ncszalen@cyfronet.pl.
authors.forename: M
authors.initials: M
authors.lastname: Szaleniec
mesh_terms: D000438:Alcohols / Q000737:chemistry* / Q000378:metabolism*; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D005292:Ferricyanides; D006900:Hydroxylation; D060847:Metabolic Engineering / Q000379:methods*; D008982:Molybdenum; D010088:Oxidoreductases / Q000737:chemistry / Q000378:metabolism*; D043362:Rhodocyclaceae / Q000201:enzymology; D013237:Stereoisomerism; D013379:Substrate Specificity
chemical_list: D000438:Alcohols; D004800:Enzymes, Immobilized; D005292:Ferricyanides; D008982:Molybdenum; D010088:Oxidoreductases; C121602:ethylbenzene dehydrogenase; C028033:potassium ferricyanide
keywords: Chiral alcohols; Enzyme immobilization; Ethylbenzene dehydrogenase; Hydroxylation