Discovery of a diverse set of esterases from hot spring microbial mat and sea sediment metagenomes

Mukhia

Front. Bioeng. Biotechnol.

et al. 2020

Bacterial lipases with activity spanning over a broad temperature and substrate range have several industrial applications. An efficient enzyme-producing bacterium Chryseobacterium polytrichastri ERMR1:04, previously reported from Sikkim Himalaya, was explored for purification and characterization of cold-adapted lipase. Optimum lipase production was observed in 1% (v/v) rice bran oil, pH 7 at 20 • C. Size exclusion and hydrophobic interaction chromatography purified the enzyme up to 21.3-fold predicting it to be a hexameric protein of 250 kDa, with 39.8 kDa monomeric unit. MALDI-TOF-MS analysis of the purified lipase showed maximum similarity with alpha/beta hydrolase (lipase superfamily). Biochemical characterization of the purified enzyme revealed optimum pH (8.0), temperature (37 • C) and activity over a temperature range of 5-65 • C. The tested metals (except Cu 2+ and Fe 2+) enhanced the enzyme activity and it was tolerant to 5% (v/v) methanol and isopropanol. The Km and Vmax values were determined as 0.104 mM and 3.58 U/mg, respectively for p-nitrophenyl palmitate. Bioinformatics analysis also supported in vitro findings by predicting enzyme's broad temperature and substrate specificity. The compatibility of the purified lipase with regular commercial detergents, coupled with its versatile temperature and substrate range, renders the given enzyme a promising biocatalyst for potential detergent formulations.

Section: Introductionmentioning

confidence: 99%

A Broad Temperature Active Lipase Purified From a Psychrotrophic Bacterium of Sikkim Himalaya With Potential Application in Detergent Formulation

Mukhia

Front. Bioeng. Biotechnol.

et al. 2020

Letters in Applied Microbiology

“…Cd 2+ , Zn 2+ ) out of the body and can remove these ions to the cytoplasm and reduce them to less toxic valence states, such as Hg 2+ and Cu 2+ (Silver and Phung 1996; Ranjan et al . 2018).…”

Section: Resultsmentioning

confidence: 99%

“…As for the mechanism of how microbial communities adapt to the heavy metal-polluted environment, microbes can directly transport heavy metal ions (e.g. Cd 2+ , Zn 2+ ) out of the body and can remove these ions to the cytoplasm and reduce them to less toxic valence states, such as Hg 2+ and Cu 2+ (Silver and Phung 1996;Ranjan et al 2018).…”

Section: Resultsmentioning

confidence: 99%

Diversity and functions of microbes in surface sediments under heavy metal pollution of western Chaohu Lake

Shen

Wang

Liang

et al. 2022

Significance and Impact of this Study: We analysed the effects of heavy metals in sediments from three sampling areas of Chaohu Lake on microbial communities. In this study, the evidence suggests that high concentrations of heavy metals can inhibit microbial species diversity, microorganisms related to the oxidation/reduction of heavy metals in the number of genes have been suppressed. Based on the type and number of viruses, we speculated that the virus in transferring genes associated with oxidation/reduction of heavy metals to different microbes played a potential role. These analyses will provide new insights into the adaptation of microorganisms to environments contaminated by heavy metals.

Appl Microbiol Biotechnol

“…For an extremely thermostable esterase of Pyrococcus furiosus , Pf_Est, Mandelli et al reported K M and V max values of 0.53 mmol/L and 6.5 × 10 −3 U respectively, when the substrate was p -nitrophenyl palmitate ( p NPP) (Mandelli et al 2016 ). Compared to esterases from functional metagenomic studies, for LOB4Est, MLC3 and SLC5 esterases, López-López et al and Ranjan et al, respectively, reported K M and V max values of 0.298 mM and 263.778 U/L, 134 μM and 14.55 μmol/min/mg and 196.5 μM and 2.393 μmol/min/mg when the substrate was firstly p -nitrophenyl laurate and p -nitrophenyl butyrate (López-López et al 2015 ; Ranjan et al 2018 ). Regarding the esterases in particular, functional at temperatures around 80 °C, as is the case for our study, the kinetic parameters of Est1 are 3.0 M for K M and 31.2 U/mg for V max (Lu et al 2019 ).…”

Section: Discussionmentioning

confidence: 99%

“…From metagenomic studies, the literature reports a wide variety of esterases, resulting from microbial communities of very diverse origins such as soil (Lu et al 2019 ; Lim et al 2020 ) and hot springs (Ranjan et al 2018 ; Sharma et al 2020 ).…”

Section: Introductionmentioning

confidence: 99%

AMWEst, a new thermostable and detergent-tolerant esterase retrieved from the Albian aquifer

Adjeroud,

Kecha,

Escuder-Rodríguez

et al. 2024

A fosmid library was constructed with the metagenomic DNA from the high-temperature sediment-rich water of the Albian aquifer (Algeria). Functional screening of this library was subsequently done looking for genes encoding lipolytic enzymes. We identified a novel gene named AMWEst (1209 base pairs) encoding a protein of 402 amino acids with a predicted molecular weight of 43.44 kDa and conferring esterase activity. AMWEst was successfully overexpressed in the yeast mesophilic host Saccharomyces cerevisiae, and the expression system used proved to be efficient and produced sufficient activity for its biochemical characterization. Multiple sequence alignment indicated that AMWEst contained a conserved pentapeptide motif (Gly120-His121-Ser122-Gln123-Gly124). The optimum pH and temperature of the recombinant esterase AMWEst were 8 and 80 °C, respectively. Additionally, AMWEst showed higher activity towards short carbon substrates and showed maximum activity for p-nitrophenyl hexanoate (C6). Notably, AMWEst has a remarkable thermostability, and the enzyme retains almost maximum activity at 70 °C after incubation for 1 h. Moreover, enzyme activity was enhanced by high concentrations of SDS and Triton X-100 detergents. Key points • A novel thermostable esterase has been retrieved through functional metagenomics • The esterase is detergent-tolerant, which is attractive for some applications • The esterase can be expressed in a yeast mesophilic host to enhance its yield