Heterologous expression, purification and characterization of arylacetonitrilases from Nectria haematococca and Arthroderma benhamiae

0395703 - MBÚ 2014 RIV GB eng J - Článek v odborném periodiku
Veselá, Alicja Barbara - Petříčková, Alena - Weyrauch, P. - Martínková, Ludmila
Heterologous expression, purification and characterization of arylacetonitrilases from Nectria haematococca and Arthroderma benhamiae.
Biocatalysis and Biotransformation. Roč. 31, č. 1 (2013), s. 49-56. ISSN 1024-2422. E-ISSN 1029-2446
Grant CEP: GA ČR(CZ) GAP504/11/0394; GA TA ČR TA01021368; GA ČR GD305/09/H008; GA MŠMT OC09046
Institucionální podpora: RVO:61388971
Klíčová slova: Arthroderma benhamiae * Arylacetonitrilase * Nectria haematococca
Kód oboru RIV: CE - Biochemie
Impakt faktor: 1.093, rok: 2013

Two novel arylacetonitrilases were purified from Escherichia coli BL21-Gold (DE3) expressing nit genes from Nectria haematococca mpVI 77-13-4 (EEU45207; NitNh) and Arthroderma benhamiae CBS 112371 (EFE30690; NitAb). The nitrilases formed holoenzymes of 360 and 336 kDa through gel filtration, while their apparent subunit size in SDS-PAGE was approximately 36 and 37 kDa, respectively. The preferred substrates of the purified enzymes were phenylacetonitrile, (R, S)-mandelonitrile, and 3-indolylacetonitrile. Both enzymes hydrolyzed (R)-mandelonitrile preferentially but with different degrees of selectivity, the e.e.s of the product (R)-mandelic acid being 63 and 89% in NitAb and NitNh, respectively, at pH 8.0. NitAb exhibited a higher temperature and pH stability than NitNh. Significant amounts of amide (> 5% of total product) were produced only by NitNh (from 2-cyanopyridine, (R, S)-mandelonitrile and phenylacetonitrile)
Trvalý link: http://hdl.handle.net/11104/0223680