Metagenome-derived Haloalkane Dehalogenases with Novel
Catalytic Properties

J 2017

Metagenome-derived Haloalkane Dehalogenases with Novel Catalytic Properties

KOTÍK, Martin, Pavel VAŇÁČEK, Antonín KUNKA, Zbyněk PROKOP, Jiří DAMBORSKÝ et. al.

Základní údaje

Originální název

Metagenome-derived Haloalkane Dehalogenases with Novel Catalytic Properties

Autoři

KOTÍK, Martin (203 Česká republika), Pavel VAŇÁČEK (203 Česká republika, domácí), Antonín KUNKA (203 Česká republika, domácí), Zbyněk PROKOP (203 Česká republika, domácí) a Jiří DAMBORSKÝ (203 Česká republika, garant, domácí)

Vydání

APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, New York, Springer, 2017, 0175-7598

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

20803 Environmental biotechnology related ethics

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 3.340

Kód RIV

RIV/00216224:14310/17:00095407

Organizační jednotka

Přírodovědecká fakulta

DOI

http://dx.doi.org/10.1007/s00253-017-8393-3

UT WoS

000407077400009

Klíčová slova anglicky

Haloalkane dehalogenase; Metagenomic DNA; Heterologous production; Substrate specificity; Protein stability

Štítky

NZ, rivok

Změněno: 9. 4. 2018 10:36, Ing. Nicole Zrilić

Anotace

V originále

Haloalkane dehalogenases (HLDs) are environmentally relevant enzymes cleaving a carbon-halogen bond in a wide range of halogenated pollutants. PCR with degenerate primers and genome-walking was used for the retrieval of four HLD-encoding genes from groundwater-derived environmental DNA. Using specific primers and the environmental DNA as a template, we succeeded in generating additional amplicons, resulting altogether in three clusters of sequences with each cluster comprising 8–13 closely related putative HLD-encoding genes. A phylogenetic analysis of the translated genes revealed that three HLDs are members of the HLD-I subfamily, whereas one gene encodes an enzyme from the subfamily HLD-II. Two metagenome-derived HLDs, eHLDB and eHLD-C, each from a different subfamily, were heterologously produced in active form, purified and characterized in terms of their thermostability, pH and temperature optimum, quaternary structure, substrate specificity towards 30 halogenated compounds, and enantioselectivity. eHLD-B and eHLD-C showed striking differences in their activities, substrate preferences, and tolerance to temperature. Profound differences were also determined in the enantiopreference and enantioselectivity of these enzymes towards selected substrates. Comparing our data with those of known HLDs revealed that eHLD-C exhibits a unique combination of high thermostability, high activity, and an unusually broad pH optimum, which covers the entire range of pH 5.5–8.9. Moreover, a so far unreported high thermostability for HLDs was determined for this enzyme at pH values lower than 6.0. Thus, eHLD-C represents an attractive and novel biocatalyst for biotechnological applications.

Návaznosti

GA16-07965S, projekt VaV

Název: Řízená evoluce dynamických elementů v enzymech s využitím mikrofluidních čipů

Investor: Grantová agentura ČR, Řízená evoluce dynamických elementů v enzymech s využitím mikrofluidních čipů

LM2015047, projekt VaV

Název: Česká národní infrastruktura pro biologická data (Akronym: ELIXIR-CZ)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Česká národní infrastruktura pro biologická data

LM2015051, projekt VaV

Název: Centrum pro výzkum toxických látek v prostředí (Akronym: RECETOX RI)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Výzkumná infrastruktura RECETOX

LM2015055, projekt VaV

Název: Centrum pro systémovou biologii (Akronym: C4SYS)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, The national infrastructure C4SYS - Centre for Systems Biology

LO1214, projekt VaV

Název: Centrum pro výzkum toxických látek v prostředí (Akronym: RECETOX)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Centrum pro výzkum toxických látek v prostředí

Citovat

KOTÍK, Martin, Pavel VAŇÁČEK, Antonín KUNKA, Zbyněk PROKOP a Jiří DAMBORSKÝ. Metagenome-derived Haloalkane Dehalogenases with Novel Catalytic Properties. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY. New York: Springer, 2017, roč. 101, č. 16, s. 6385-6397. ISSN 0175-7598. Dostupné z: https://dx.doi.org/10.1007/s00253-017-8393-3.

@article{1403838,
   author = {Kotík, Martin and Vaňáček, Pavel and Kunka, Antonín and Prokop, Zbyněk and Damborský, Jiří},
   article_location = {New York},
   article_number = {16},
   doi = {http://dx.doi.org/10.1007/s00253-017-8393-3},
   keywords = {Haloalkane dehalogenase; Metagenomic DNA; Heterologous production; Substrate specificity; Protein stability},
   language = {eng},
   issn = {0175-7598},
   journal = {APPLIED MICROBIOLOGY AND BIOTECHNOLOGY},
   title = {Metagenome-derived Haloalkane Dehalogenases with Novel Catalytic Properties},
   url = {https://loschmidt.chemi.muni.cz/peg/publications/metagenome-derived-haloalkane-dehalogenases-with-novel-catalytic-properties/},
   volume = {101},
   year = {2017}
}

TY  - JOUR
ID  - 1403838
AU  - Kotík, Martin - Vaňáček, Pavel - Kunka, Antonín - Prokop, Zbyněk - Damborský, Jiří
PY  - 2017
TI  - Metagenome-derived Haloalkane Dehalogenases with Novel Catalytic Properties
JF  - APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
VL  - 101
IS  - 16
SP  - 6385-6397
EP  - 6385-6397
PB  - Springer
SN  - 01757598
KW  - Haloalkane dehalogenase
KW  - Metagenomic DNA
KW  - Heterologous production
KW  - Substrate specificity
KW  - Protein stability
UR  - https://loschmidt.chemi.muni.cz/peg/publications/metagenome-derived-haloalkane-dehalogenases-with-novel-catalytic-properties/
N2  - Haloalkane dehalogenases (HLDs) are environmentally relevant enzymes cleaving a carbon-halogen bond in a wide range of halogenated pollutants. PCR with degenerate primers and genome-walking was used for the retrieval of four HLD-encoding genes from groundwater-derived environmental DNA. Using specific primers and the environmental DNA as a template, we succeeded in generating additional amplicons, resulting altogether in three clusters of sequences with each cluster comprising 8–13 closely related putative HLD-encoding genes. A phylogenetic analysis of the translated genes revealed that three HLDs are members of the HLD-I subfamily, whereas one gene encodes an enzyme from the subfamily HLD-II. Two metagenome-derived HLDs, eHLDB and eHLD-C, each from a different subfamily, were heterologously produced in active form, purified and characterized in terms of their thermostability, pH and temperature optimum, quaternary structure, substrate specificity towards 30 halogenated compounds, and enantioselectivity. eHLD-B and eHLD-C showed striking differences in their activities, substrate preferences, and tolerance to temperature. Profound differences were also determined in the enantiopreference and enantioselectivity of these enzymes towards selected substrates. Comparing our data with those of known HLDs revealed that eHLD-C exhibits a unique combination of high thermostability, high activity, and an unusually broad pH optimum, which covers the entire range of pH 5.5–8.9. Moreover, a so far unreported high thermostability for HLDs was determined for this enzyme at pH values lower than 6.0. Thus, eHLD-C represents an attractive and novel biocatalyst for biotechnological applications.
ER  -

KOTÍK, Martin, Pavel VAŇÁČEK, Antonín KUNKA, Zbyněk PROKOP a Jiří DAMBORSKÝ. Metagenome-derived Haloalkane Dehalogenases with Novel Catalytic Properties. \textit{APPLIED MICROBIOLOGY AND BIOTECHNOLOGY}. New York: Springer, 2017, roč.~101, č.~16, s.~6385-6397. ISSN~0175-7598. Dostupné z: https://dx.doi.org/10.1007/s00253-017-8393-3.

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