Biochemical profiles of two thermostable and organic solvent-tolerant esterases derived from a compost metagenome

Applied Microbiology and Biotechnology
Mingji LuRolf Daniel

Abstract

Owing to the functional versatility and potential applications in industry, interest in lipolytic enzymes tolerant to organic solvents is increasing. In this study, functional screening of a compost soil metagenome resulted in identification of two lipolytic genes, est1 and est2, encoding 270 and 389 amino acids, respectively. The two genes were heterologously expressed and characterized. Est1 and Est2 are thermostable enzymes with optimal enzyme activities at 80 and 70 °C, respectively. A second-order rotatable design, which allows establishing the relationship between multiple variables with the obtained responses, was used to explore the combined effects of temperature and pH on esterase stability. The response curve indicated that Est1, and particularly Est2, retained high stability within a broad range of temperature and pH values. Furthermore, the effects of organic solvents on Est1 and Est2 activities and stabilities were assessed. Notably, Est2 activity was significantly enhanced (two- to tenfold) in the presence of ethanol, methanol, isopropanol, and 1-propanol over a concentration range between 6 and 30% (v/v). For the short-term stability (2 h of incubation), Est2 exhibited high tolerance against 60% (v/v) of ethanol...Continue Reading

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Datasets Mentioned

BETA
KR149567.1
KR149568.1
ETW96815
EMF58012

Methods Mentioned

BETA
PCR
electrophoresis

Software Mentioned

Clone Manager
FramePlot
BLASTP
MEGA
ESPript3
SignalP
Expresso webserver
TASSER

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