Plant Nitrilase Homologues in Fungi: Phylogenetic and Functional Analysis with Focus on Nitrilases in Trametes versicolor and Agaricus bisporus

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F20%3A00532967" target="_blank" >RIV/61388971:_____/20:00532967 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/68407700:21460/20:00348820

Výsledek na webu

<a href="https://www.mdpi.com/1420-3049/25/17/3861" target="_blank" >https://www.mdpi.com/1420-3049/25/17/3861</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.3390/molecules25173861" target="_blank" >10.3390/molecules25173861</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Plant Nitrilase Homologues in Fungi: Phylogenetic and Functional Analysis with Focus on Nitrilases in Trametes versicolor and Agaricus bisporus

Popis výsledku v původním jazyce

Fungi contain many plant-nitrilase (NLase) homologues according to database searches. In this study, enzymes NitTv1 fromTrametes versicolorand NitAb fromAgaricus bisporuswere purified and characterized as the representatives of this type of fungal NLase. Both enzymes were slightly more similar to NIT4 type than to NIT1/NIT2/NIT3 type of plant NLases in terms of their amino acid sequences. Expression of the synthetic genes inEscherichia coliOrigami B (DE3) was induced with 0.02 mM isopropyl beta-D-1-thiogalactopyranoside at 20 degrees C. Purification of NitTv1 and NitAb by cobalt affinity chromatography gave ca. 6.6 mg and 9.6 mg of protein per 100 mL of culture medium, respectively. Their activities were determined with 25 mM of nitriles in 50 mM Tris/HCl buffer, pH 8.0, at 30 degrees C. NitTv1 and NitAb transformed beta-cyano-L-alanine (beta-CA) with the highest specific activities (ca. 132 and 40 U mg(-1), respectively) similar to plant NLase NIT4. beta-CA was transformed into Asn and Asp as in NIT4 but at lower Asn:Asp ratios. The fungal NLases also exhibited significant activities for (aryl)aliphatic nitriles such as 3-phenylpropionitrile, cinnamonitrile and fumaronitrile (substrates of NLase NIT1). NitTv1 was more stable than NitAb (at pH 5-9 vs. pH 5-7). These NLases may participate in plant-fungus interactions by detoxifying plant nitriles and/or producing plant hormones. Their homology models elucidated the molecular interactions with various nitriles in their active sites.

Název v anglickém jazyce

Plant Nitrilase Homologues in Fungi: Phylogenetic and Functional Analysis with Focus on Nitrilases in Trametes versicolor and Agaricus bisporus

Popis výsledku anglicky

Fungi contain many plant-nitrilase (NLase) homologues according to database searches. In this study, enzymes NitTv1 fromTrametes versicolorand NitAb fromAgaricus bisporuswere purified and characterized as the representatives of this type of fungal NLase. Both enzymes were slightly more similar to NIT4 type than to NIT1/NIT2/NIT3 type of plant NLases in terms of their amino acid sequences. Expression of the synthetic genes inEscherichia coliOrigami B (DE3) was induced with 0.02 mM isopropyl beta-D-1-thiogalactopyranoside at 20 degrees C. Purification of NitTv1 and NitAb by cobalt affinity chromatography gave ca. 6.6 mg and 9.6 mg of protein per 100 mL of culture medium, respectively. Their activities were determined with 25 mM of nitriles in 50 mM Tris/HCl buffer, pH 8.0, at 30 degrees C. NitTv1 and NitAb transformed beta-cyano-L-alanine (beta-CA) with the highest specific activities (ca. 132 and 40 U mg(-1), respectively) similar to plant NLase NIT4. beta-CA was transformed into Asn and Asp as in NIT4 but at lower Asn:Asp ratios. The fungal NLases also exhibited significant activities for (aryl)aliphatic nitriles such as 3-phenylpropionitrile, cinnamonitrile and fumaronitrile (substrates of NLase NIT1). NitTv1 was more stable than NitAb (at pH 5-9 vs. pH 5-7). These NLases may participate in plant-fungus interactions by detoxifying plant nitriles and/or producing plant hormones. Their homology models elucidated the molecular interactions with various nitriles in their active sites.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2020

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Molecules

ISSN

1420-3049

e-ISSN

—

Svazek periodika

25

Číslo periodika v rámci svazku

17

Stát vydavatele periodika

CH - Švýcarská konfederace

Počet stran výsledku

20

Strana od-do

3861

Kód UT WoS článku

000570398700001

EID výsledku v databázi Scopus

2-s2.0-85090004951