Identification and characterization of a novel carboxylesterase EstQ7 from a soil metagenomic library

© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature..

A novel lipolytic gene, estq7, was identified from a fosmid metagenomic library. The recombinant enzyme EstQ7 consists of 370 amino acids with an anticipated molecular mass of 42 kDa. Multiple sequence alignments showed that EstQ7 contained a pentapeptide motif GHSMG, and a putative catalytic triad Ser174-Asp306-His344. Interestingly, EstQ7 was found to have very little similarity to the characterized lipolytic enzymes. Phylogenetic analysis revealed that EstQ7 may be a member of a novel family of lipolytic enzymes. Biochemical characterization of the recombinant enzyme revealed that it constitutes a slightly alkalophilic, moderate thermophilic and highly active carboxylesterase against short-chain fatty acid esters with optimum temperature 50 ℃ and pH 8.2. The Km and kcat values toward p-nitrophenyl acetate were determined to be 0.17 mM and 1910s-1, respectively. Moreover, EstQ7 was demonstrated to have acyltransferase activity by GC-MS analysis. Structural modeling of the three-dimensional structure of this new enzyme showed that it exhibits a typical α/β hydrolase fold, and the catalytic triad residues are spatially close. Molecular docking revealed the interactions between the enzyme and the ligand. The high levels of lipolytic activity of EstQ7, combined with its moderate thermophilic property and acyltransferase activity, render this novel enzyme a promising candidate biocatalyst for food, pharmaceutical and biotechnological applications.

Medienart:

E-Artikel

Erscheinungsjahr:

2021

Erschienen:

2021

Enthalten in:

Zur Gesamtaufnahme - volume:203

Enthalten in:

Archives of microbiology - 203(2021), 7 vom: 21. Sept., Seite 4113-4125

Sprache:

Englisch

Beteiligte Personen:

Yan, Zhenzhen [VerfasserIn]
Ding, Liping [VerfasserIn]
Zou, Dandan [VerfasserIn]
Wang, Luyao [VerfasserIn]
Tan, Yuzhi [VerfasserIn]
Guo, Shuting [VerfasserIn]
Zhang, Yingchen [VerfasserIn]
Xin, Zhihong [VerfasserIn]

Links:

Volltext

Themen:

Acyltransferase
Carboxylesterase
EC 3.1.1.1
Journal Article
Metagenome
Molecular docking
Unculturable microorganism

Anmerkungen:

Date Completed 17.09.2021

Date Revised 17.09.2021

published: Print-Electronic

Citation Status MEDLINE

doi:

10.1007/s00203-021-02398-0

funding:

Förderinstitution / Projekttitel:

PPN (Katalog-ID):

NLM326061762