Metagenomic approach for the isolation of a novel low-temperature-active lipase from uncultured bacteria of marine sediment

Hårdeman, Fredrik; Sjöling, Sara

doi:10.1111/j.1574-6941.2006.00206.x

Cited by 154 publications

(99 citation statements)

References 61 publications

(123 reference statements)

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“…These include low substrate affinity, thermolability, and high specific activity at low temperatures, which can together help achieve saving in energy costs and in reducing undesirable chemical side reactions; as well as allowing rapid thermal inactivation (Cavicchioli et al, 2002;Santiago et al, 2016). Other "cold-active" lipolytic enzymes from family IV have previously been described, but these usually possess higher optimal temperatures (35-50 • C; Hårdeman and Sjöling, 2007;Fu et al, 2011), whereas our esterase has a high activity at 4-40 • C, identifying it as truly "cold-active." In addition this work also broadens the description of members of the lipolytic enzyme family IV, as thermophilic and mesophilic enzymes of this family have to date been already described (Rhee et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

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: Introductionmentioning

confidence: 99%

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

et al. 2017

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“…Using this approach, several groups isolated novel enzymes from various environments (Ranjan et al, 2005;Rhee et al, 2005;Feng et al, 2007;Hardeman and Sjoling, 2007;Tirawongsaroj et al, 2008), and many of them were found to offer a good potential as new tools for industrial applications, because their biophysical properties arise from improved adaptations to the conditions met in the environment during the sampling process. Although such studies reported the characterization of metagenome-derived enzymes, insights into their physiological aspect are often lacking.…”

Section: Introductionmentioning

confidence: 99%

Insights into bacterial cellulose biosynthesis by functional metagenomics on Antarctic soil samples

et al. 2009

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In this study, the mining of an Antarctic soil sample by functional metagenomics allowed the isolation of a cold-adapted protein (RBcel1) that hydrolyzes only carboxymethyl cellulose. The new enzyme is related to family 5 of the glycosyl hydrolase (GH5) protein from Pseudomonas stutzeri (Pst_2494) and does not possess a carbohydrate-binding domain. The protein was produced and purified to homogeneity. RBcel1 displayed an endoglucanase activity, producing cellobiose and cellotriose, using carboxymethyl cellulose as a substrate. Moreover, the study of pH and the thermal dependence of the hydrolytic activity shows that RBcel1 was active from pH 6 to pH 9 and remained significantly active when temperature decreased (18% of activity at 10 1C). It is interesting that RBcel1 was able to synthetize non-reticulated cellulose using cellobiose as a substrate. Moreover, by a combination of bioinformatics and enzyme analysis, the physiological relevance of the RBcel1 protein and its mesophilic homologous Pst_2494 protein from P. stutzeri, A1501, was established as the key enzymes involved in the production of cellulose by bacteria. In addition, RBcel1 and Pst_2494 are the two primary enzymes belonging to the GH5 family involved in this process.

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“…Reasons for this include the reluctance of many microorganisms present in these samples to lyse by protocols that have been developed mainly for DNA extraction from mesophilic samples and the release of very stable nucleases upon cell lysis. Nevertheless, significant progress has been made, and various methods allowing the isolation of highquality DNA from a variety of environments, i.e., soil (45,87,134,139), marine picoplankton (117), contaminated subsurface sediments (1), groundwater (128), hot springs and mud holes in solfataric fields (94), surface water from rivers (145), glacier ice (109), Antarctic desert soil (48), and buffalo rumens (25), have been developed.…”

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confidence: 99%

Metagenomic Analyses: Past and Future Trends

Simon

Daniel

2011

Appl Environ Microbiol

631

349

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Metagenomics has revolutionized microbiology by paving the way for a cultivation-independent assessment and exploitation of microbial communities present in complex ecosystems. Metagenomics comprising construction and screening of metagenomic DNA libraries has proven to be a powerful tool to isolate new enzymes and drugs of industrial importance. So far, the majority of the metagenomically exploited habitats comprised temperate environments, such as soil and marine environments. Recently, metagenomes of extreme environments have also been used as sources of novel biocatalysts. The employment of next-generation sequencing techniques for metagenomics resulted in the generation of large sequence data sets derived from various environments, such as soil, the human body, and ocean water. Analyses of these data sets opened a window into the enormous taxonomic and functional diversity of environmental microbial communities. To assess the functional dynamics of microbial communities, metatranscriptomics and metaproteomics have been developed. The combination of DNA-based, mRNA-based, and protein-based analyses of microbial communities present in different environments is a way to elucidate the compositions, functions, and interactions of microbial communities and to link these to environmental processes.

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Metagenomic approach for the isolation of a novel low-temperature-active lipase from uncultured bacteria of marine sediment

Cited by 154 publications

References 61 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

Insights into bacterial cellulose biosynthesis by functional metagenomics on Antarctic soil samples

Metagenomic Analyses: Past and Future Trends

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