Lipases and Esterases from Extremophiles: Overview and Case Example of the Production and Purification of an Esterase from Thermus thermophilus HB27

Fuciños, Pablo; González, Roberto; Atanes, Estrella; Sestelo, A.B.F.; Guerra, Nelson Pérez; Pastrana, Lorenzo; Rúa, M. Luisa

doi:10.1007/978-1-61779-600-5_15

Cited by 38 publications

(21 citation statements)

References 76 publications

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“…Resilience against organic solvents is of particular interest for industrial applications, as esterases are widely used in processes like the synthesis of pharmaceutical and fine chemicals where these solvents are present (Fuciños et al, 2012). All tested organic solvents showed an inhibiting effect on ML-005.…”

Section: Resultsmentioning

confidence: 99%

Characterization of ML-005, a Novel Metaproteomics-Derived Esterase

2018

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A novel gene encoding for a lipolytic enzyme, designated ML-005, was recently identified using a functional metaproteomics approach. We heterologously expressed this protein in Escherichia coli and biochemically characterized it. ML-005 exhibited lipolytic activity toward short-chained substrates with the preferred substrate being p-nitrophenyl-butyrate, suggesting that ML-005 is an esterase. According to homology analysis and site-directed mutagenesis, the catalytic triad of the enzyme was identified as Ser-99, Asp-164, and His-191. Its optimal pH was determined to be at pH 8. Optimal activity was observed at 45°C. It also exhibited temperature, pH and salt tolerance. Residual relative activity after incubating at 50–60°C for 360 min was above 80% of its initial activity. It showed tolerance over a broad range of pH (5–12) and retained most of its initial activity. Furthermore, incubating ML-005 in 1 – 5M NaCl solution had negligible effect on its activity. DTT, EDTA, and ß-mercaptoethanol had no significant effect on ML-005’s activity. However, addition of PMSF led to almost complete inactivation consistent with ML-005 being a serine hydrolase. ML-005 remains stable in the presence of a range of metal ions, but addition of Cu2+ significantly reduces its relative activity. Organic solvents have an inhibitory effect on ML-005, but it retained 21% of activity in 10% methanol. SDS had the most pronounced inhibitory effect on ML-005 among all detergents tested and completely inactivated it. Furthermore, the Vmax of ML-005 was determined to be 59.8 μM/min along with a Km of 137.9 μM. The kcat of ML-005 is 26 s-1 and kcat/Km is 1.88 × 105 M-1 s-1.

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Section: Resultsmentioning

confidence: 99%

Characterization of ML-005, a Novel Metaproteomics-Derived Esterase

2018

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“…Besides, the purified esterase also showed better activity against C6 (pNPH), C8 (pNPC), C10 (pNPD) and C12 (pNPL). The reported esterases from deep-sea metagenomic library [38], Oenococcus oeni [39], Geobacillus thermodenitrificans T2 [40], Thermus thermophilus HB27 [13] had also shown a strong preference for the hydrolysis of pNPB. However the present esterase with broad substrate specificity offers potential applications in the production of various high value products.…”

Section: Chemicalsmentioning

confidence: 89%

“…With respect to this fact enzymes from extremophiles often perform better in various commercial applications compared to normal biocatalysts [14]. So far numerous organic solvent tolerant, halo-tolerant and thermostable esterases have been reported individually [13,15]. Recently we reported a thermostable, organic solvent tolerant and alkali tolerant esterase produced by a thermophilic bacteria Geobacillus sp.…”

Section: Introductionmentioning

confidence: 99%

“…They are important due to their stability under high temperature conditions [13]. At higher temperature, catalytic rates of enzyme increases, even in some cases thermophilic enzymes retain organic solvent stability, making them important biocatalysts for large scale organic syntheses or reactions where reduced water activity is quite desired.…”

Section: Introductionmentioning

confidence: 99%

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Purification and characterization of a thermo-halophilic, alkali-stable and extremely benzene tolerant esterase from a thermo-halo tolerant Bacillus cereus strain AGP-03, isolated from ‘Bakreshwar’ hot spring, India

Ghati

Paul

2015

Process Biochemistry

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“…Harsh, extremophilic environments can be sources of biotechnologically relevant bacteria and therefore hold great promise for the biotechnological industry (Raddadi et al, 2015). For example, extremophilic microorganisms can yield enzymes such as lipases and esterases that can be used under a wide range of conditions and may have relevant applications in the food, detergent and biofuel industries (Fuciños et al, 2012). There are many other examples of biotechnologically relevant microorganisms from extreme environments, including the wellknown case of Thermus aquaticus, which produces the widely used Taq polymerase; or the hyperthermophilic biofuel-producing archaea that live in deep-sea hydrothermal vents (Chien et al, 1976;Nishimura and Sako, 2009).…”

Section: Introductionmentioning

confidence: 99%

Microbial communities of the Mediterranean rocky shore: ecology and biotechnological potential of the sea‐land transition

Molina‐Menor

Tanner

Vidal‐Verdú

et al. 2019

Microbial Biotechnology

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Summary Microbial communities from harsh environments hold great promise as sources of biotechnologically relevant strains and compounds. In the present work, we have characterized the microorganisms from the supralittoral and splash zone in three different rocky locations of the Western Mediterranean coast, a tough environment characterized by high levels of irradiation and large temperature and salinity fluctuations. We have retrieved a complete view of the ecology and functional aspects of these communities and assessed the biotechnological potential of the cultivable microorganisms. All three locations displayed very similar taxonomic profiles, with the genus Rubrobacter and the families Xenococcaceae, Flammeovirgaceae, Phyllobacteriaceae, Rhodobacteraceae and Trueperaceae being the most abundant taxa; and Ascomycota and halotolerant archaea as members of the eukaryotic and archaeal community respectively. In parallel, the culture‐dependent approach yielded a 100‐isolates collection, out of which 12 displayed high antioxidant activities, as evidenced by two in vitro (hydrogen peroxide and DPPH) and confirmed in vivo with Caenorhabditis elegans assays, in which two isolates, CR22 and CR24, resulted in extended survival rates of the nematodes. This work is the first complete characterization of the Mediterranean splash‐zone coastal microbiome, and our results indicate that this microbial niche is home of an extremophilic community that holds biotechnological potential.

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Lipases and Esterases from Extremophiles: Overview and Case Example of the Production and Purification of an Esterase from Thermus thermophilus HB27

Cited by 38 publications

References 76 publications

Characterization of ML-005, a Novel Metaproteomics-Derived Esterase

Characterization of ML-005, a Novel Metaproteomics-Derived Esterase

Purification and characterization of a thermo-halophilic, alkali-stable and extremely benzene tolerant esterase from a thermo-halo tolerant Bacillus cereus strain AGP-03, isolated from ‘Bakreshwar’ hot spring, India

Microbial communities of the Mediterranean rocky shore: ecology and biotechnological potential of the sea‐land transition

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