title: Production of epoxide hydrolases in batch fermentations of Botryosphaeria rhodina.
abstract: The filamentous fungus Botryosphaeria rhodina (ATCC 9055) was investigated related to its ability for epoxide hydrolase (EH) production. Epoxide hydrolase activity is located at two different sites of the cells. The larger part is present in the cytosol (70%), while the smaller part is associated to membranes (30%). In media optimization experiments, an activity of 3.5 U/gDW for aromatic epoxide hydrolysis of para-nitro-styrene oxide (pNSO) could be obtained. Activity increased by 30% when pNSO was added to the culture during exponential growth. An increase of enzyme activity up to 6 U/gDW was achieved during batch-fermentations in a bioreactor with 2.7 l working volume. Evaluation of fermentations with 30 l working volume revealed a relation of oxygen uptake rate to EH expression. Oxygen limitation resulted in a decreased EH activity. Parameter estimation by the linearization method of Hanes yielded Km values of 2.54 and 1.00 mM for the substrates S-pNSO and R-pNSO, respectively. vmax was 3.4 times higher when using R-pNSO. A protein purification strategy leading to a 47-fold increase in specific activity (940 U/mgProtein) was developed as a first step to investigate molecular and structural characteristics of the EH.
authors.affiliation: Institute of Biochemical Engineering, Technische Universität Braunschweig, Gaussstrasse 17, 38106 Braunschweig, Germany.
authors.forename: Guido
authors.initials: G
authors.lastname: Melzer
authors.forename: Stefan
authors.initials: S
authors.lastname: Junne
authors.forename: Roland
authors.initials: R
authors.lastname: Wohlgemuth
authors.forename: Dietmar C
authors.initials: DC
authors.lastname: Hempel
authors.forename: Peter
authors.initials: P
authors.lastname: Götz
mesh_terms: D001203:Ascomycota / Q000187:drug effects / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism; D019149:Bioreactors; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D004851:Epoxide Hydrolases / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism*; D005285:Fermentation; D007477:Ions / Q000494:pharmacology; D008670:Metals / Q000494:pharmacology; D010100:Oxygen / Q000378:metabolism; D013696:Temperature
chemical_list: D003470:Culture Media; D007477:Ions; D008670:Metals; D004851:Epoxide Hydrolases; D010100:Oxygen