Original title: Nové možnosti aplikace nittrilas v biokatalýze a bioremediaci
Translated title: New possibilities of nitrilases in biocatalysis and bioremediation
Authors: Veselá, Alicja Barbara ; Bezouška, Karel (advisor) ; Weignerová, Lenka (referee)
Document type: Master’s theses
Year: 2011
Language: cze
Abstract: [cze] [eng]
Nitrilasy jsou enzymy, jež katalyzují hydrolýzu nitrilů na karboxylové kyseliny. Pro svou chemo- a enantioselektivitu mají tyto enzymy velký potenciál pro využití v biokatalýze, např. při syntéze kyseliny mandlové a mandelamidu, nebo bioremediaci míst kontaminovaných organickými nitrily. V této práci byla studována nitrilasová aktivita rekombinantních kmenů E. coli exprimujících hypotetické fungální nitrilasy z hub Giberella moniliformis a Nectria haematococca mpVI 77-13-4 a schopnost bakterií rodu Rhodococcus a Nocardia hydrolyzovat benzonitrilové herbicidy dichlobenil (2,6-dichlorbenzonitril), ioxynil (3,5-dijod- 4-hydroxybenzonitril) a bromoxynil (3,5-dibrom-4-hydroxybenzonitril). Hypotetické fungální nitrilasy byly exprimovány jako funkční enzymy. Nitrilasa z G. moniliformis vykazovala nejvyšší aktivitu k benzonitrilu (30,9 U/mg proteinu); při kultivaci byla získána aktivita 2560 U/l buněčné suspenze. Preferovaným substrátem enzymu z N. haematococca byl fenylacetonitril (12,3 U/mg prot.), celkem bylo získáno 28050 U/l buněčné suspenze. Nitrilasa z N. haematococca byla schopna hydrolyzovat mandelonitril (5,9 U/mg prot.). Půdní bakterie Rhodococcus rhodochrous PA-34, Nocardia globerula NHB-2 a Rhodococcus sp. NDB 1165 hydrolyzovaly herbicidy ioxynil, bromoxynil a chlorovaný analog 3,5-dichlor-4-... Nitrilases are enzymes which catalyze the hydrolysis of nitriles to corresponding carboxylic acids. These enzymes have a great potential in biocatalysis, for example in the synthesis of mandelic acid and mandelamide, because of their chemo- and enantioselectivity. As bioremediation agents they are also applicable to sites contaminated with organic nitriles. In this work, activities of recombinant strains of E. coli expressing hypothetical nitrilases from fungi Giberella moniliformis and Nectria haematococca mpVI 77-13-4 were studied, as well as the biodegradation potential of bacteria from Rhodococcus and Nocardia genera towards benzonitrile herbicides dichlobenil (2,6-dichlorobenzonitrile), ioxynil (3,5-diiodo-4- hydroxybenzonitrile) and bromoxynil (3,5-dibromo-4-hydroxybenzonitrile). The hypothetical fungal nitrilases were expressed as functional enzymes. Nitrilase from G. moniliformis showed highest activity towards benzonitrile (30.9 U/mg protein), total activity yield was 2,560 U/l cell culture. The preferred substrate of the nitrilase from N. haematococca was phenylacetonitrile (12.3 U/mg prot.), total activity yield was 28,050 U/l cell culture. Nitrilase from N. haematococca was also able to hydrolyze mandelonitrile (5.9 U/mg prot.). Soil bacteria Rhodococcus rhodochrous PA-34, Nocardia globerula...
Keywords: Gibberella moniliformis; Nectria haematococca; Nitrilase; Nocardia; Rhodococcus; Gibberella moniliformis; Nectria haematococca; Nitrilasa; Nocardia; Rhodococcus

Institution: Charles University Faculties (theses) (web)
Document availability information: Available in the Charles University Digital Repository.
Original record: http://hdl.handle.net/20.500.11956/33077

Permalink: http://www.nusl.cz/ntk/nusl-297553


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Universities and colleges > Public universities > Charles University > Charles University Faculties (theses)
Academic theses (ETDs) > Master’s theses