A novel, versatile family IV carboxylesterase exhibits high stability and activity in a broad pH spectrum

Dukunde, Amélie; Schneider, Dominik; Lu, Mi; Brady, Silja; Daniel, Rolf

doi:10.1007/s10529-016-2282-1

Cited by 24 publications

(12 citation statements)

References 52 publications

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“…Mechanisms involved in metal ion resistance in esterases are not well established, with different effects on enzyme activity being observed with some metal ions. For example while the Est06 esterase has been shown to be resistant to Cu 2+ (Dukunde et al, 2017), we found Cu 2+ to have a positive effect on 7N9 esterase activity as it also has on esterase activity in a cold adapted enzyme isolated from Shewanella sp. (Hang et al, 2016).…”

Section: Discussionmentioning

confidence: 60%

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A Novel Cold Active Esterase from a Deep Sea Sponge Stelletta normani Metagenomic Library

et al. 2017

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Esterases catalyze the hydrolysis of ester bonds in fatty acid esters with short-chain acyl groups. Due to the widespread applications of lipolytic enzymes in various industrial applications, there continues to be an interest in novel esterases with unique properties. Marine ecosystems have long been acknowledged as a significant reservoir of microbial biodiversity and in particular of bacterial enzymes with desirable characteristics for industrial use, such as for example cold adaptation and activity in the alkaline pH range. We employed a functional metagenomic approach to exploit the enzymatic potential of one particular marine ecosystem, namely the microbiome of the deep sea sponge Stelletta normani. Screening of a metagenomics library from this sponge resulted in the identification of a number of lipolytic active clones. One of these encoded a highly, cold-active esterase 7N9, and the recombinant esterase was subsequently heterologously expressed in Escherichia coli. The esterase was classified as a type IV lipolytic enzyme, belonging to the GDSAG subfamily of hormone sensitive lipases. Furthermore, the recombinant 7N9 esterase was biochemically characterized and was found to be most active at alkaline pH (8.0) and displays salt tolerance over a wide range of concentrations. In silico docking studies confirmed the enzyme's activity toward short-chain fatty acids while also highlighting the specificity toward certain inhibitors. Furthermore, structural differences to a closely related mesophilic E40 esterase isolated from a marine sediment metagenomics library are discussed.

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Section: Discussionmentioning

confidence: 60%

“…Interestingly a novel carboxylesterase Est06, isolated from a forest soil metagenomics library has also recently been reported to share 61% similarity with a hypothetical protein from Candidatus Entotheonella sp. TSY1 (Dukunde et al, 2017). FIGURE 7 | pH dependency of 7N9 esterase.…”

Section: Discussionmentioning

confidence: 99%

A Novel Cold Active Esterase from a Deep Sea Sponge Stelletta normani Metagenomic Library

et al. 2017

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“…Furthermore, many bHSL genes were identified through metagenomic approaches directly from environmental DNA samples [46][47][48][49][50][51][52][53][54][55][56][57][58][59][60][61][62][63][64]. These metagenomic bHSLs were isolated from soil [46,53,55,56], mud and sediment-rich water [47], environmental soils [48], forest soil [49,60,64], marine sediments [50,51,58], activated sludge [52,54], mountain soil [57], tidal flat sediment [59], surface sediment [61], and permafrost [62,63]. Interestingly, bHSL family members were shown to be selectively inhibited by 5-methoxy-3-(4-phenoxyphenyl)-3H- [1,3,4]oxadiazol-2-one [65].…”

Section: Bacterial Hormone-sensitive Lipasesmentioning

confidence: 99%

Bacterial Hormone-Sensitive Lipases (bHSLs): Emerging Enzymes for Biotechnological Applications

Kim

2017

Journal of Microbiology and Biotechnology

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Lipases are important enzymes with biotechnological applications in dairy, detergent, food, fine chemicals, and pharmaceutical industries. Specifically, hormone-sensitive lipase (HSL) is an intracellular lipase that can be stimulated by several hormones, such as catecholamine, glucagon, and adrenocorticotropic hormone. Bacterial hormone-sensitive lipases (bHSLs), which are homologous to the C-terminal domain of HSL, have α/β-hydrolase fold with a catalytic triad composed of His, Asp, and Ser. These bHSLs could be used for a wide variety of industrial applications because of their high activity, broad substrate specificity, and remarkable stability. In this review, the relationships among HSLs, the microbiological origins, the crystal structures, and the biotechnological properties of bHSLs are summarized.

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“…Similarity searches showed that protein sequences similar to Est56 were mainly identified during metagenome screenings. These comprised ELP45 isolated from a forest topsoil [66], EstC23 from mountain soil [67], Est06 from forest soil [68], and EstMY from activated sludge [69], which showed 62%, 61%, 60%, and 60% amino acid identity to Est56, respectively.…”

Section: Identification and Sequence Analysis Of A Novel Lipolytic Genementioning

confidence: 99%

A Novel Carboxylesterase Derived from a Compost Metagenome Exhibiting High Stability and Activity towards High Salinity

Daniel

2021

Genes

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Halotolerant lipolytic enzymes have gained growing interest, due to potential applications under harsh conditions, such as hypersalinity and presence of organic solvents. In this study, a lipolytic gene, est56, encoding 287 amino acids was identified by functional screening of a compost metagenome. Subsequently, the gene was heterologously expressed, and the recombinant protein (Est56) was purified and characterized. Est56 is a mesophilic (Topt 50 °C) and moderate alkaliphilic (pHopt 8) enzyme, showing high thermostability at 30 and 40 °C. Strikingly, Est56 is halotolerant as it exhibited high activity and stability in the presence of up to 4 M NaCl or KCl. Est56 also displayed enhanced stability against high temperatures (50 and 60 °C) and urea (2, 4, and 6 M) in the presence of NaCl. In addition, the recently reported halotolerant lipolytic enzymes were summarized. Phylogenetic analysis grouped these enzymes into 13 lipolytic protein families. The majority (45%) including Est56 belonged to family IV. To explore the haloadaptation of halotolerant enzymes, the amino acid composition between halotolerant and halophilic enzymes was statistically compared. The most distinctive feature of halophilic from non-halophilic enzymes are the higher content of acidic residues (Asp and Glu), and a lower content of lysine, aliphatic hydrophobic (Leu, Met and Ile) and polar (Asn) residues. The amino acid composition and 3-D structure analysis suggested that the high content of acidic residues (Asp and Glu, 12.2%) and low content of lysine residues (0.7%), as well as the excess of surface-exposed acidic residues might be responsible for the haloadaptation of Est56.

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A novel, versatile family IV carboxylesterase exhibits high stability and activity in a broad pH spectrum

Abstract: The combination of Est06 properties place it among novel biocatalysts that have potential for industrial use including low temperature applications.

Cited by 24 publications

References 52 publications

A Novel Cold Active Esterase from a Deep Sea Sponge Stelletta normani Metagenomic Library

A Novel Cold Active Esterase from a Deep Sea Sponge Stelletta normani Metagenomic Library

Bacterial Hormone-Sensitive Lipases (bHSLs): Emerging Enzymes for Biotechnological Applications

A Novel Carboxylesterase Derived from a Compost Metagenome Exhibiting High Stability and Activity towards High Salinity

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