Expression and characterization of an esterase belonging to a new family via isolation from a metagenomic library of paper mill sludge

Jia, Mei-Lu; Zhong, Xiaolin; Lin, Zhiwei; Dong, Bingxue; Li, Gang

doi:10.1016/j.ijbiomac.2019.01.025

Cited by 19 publications

(13 citation statements)

References 33 publications

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“…Overall, approximately 4.89 and 2.56 Gb of cloned compost DNA were screened, yielding 199 and 51 positive clones for compost55 and compost76, respectively. Previous studies have used various vectors such as BACs, fosmids and plasmids for function-based screening of LEs from different bioresources (Lee et al 2004; Lämmle et al 2007; Kim et al 2010; Nacke 2011; Berlemont et al 2013; Shao et al 2013; Leis et al 2015; Jia et al 2019). The hit rate to recover a lipolytic-positive clone ranged from 0.714 to 208 per Gb of cloned DNA (Table 1).…”

Section: Resultsmentioning

confidence: 99%

“…Patatin-like-proteins and tannases (designated as P and T, respectively) were shaded in yellow. Other recent reported lipolytic families were shaded in magenta: 1, Est22 (Li et al 2017); 2, EstL28 (Seo et al 2014); 3, Rv0045c (Guo et al 2010); 4, EstGX1 (Jiménez et al 2012); 5, EstLiu (Rahman et al 2016); 6, EstY (Wu and Sun 2009); 7; EstGS (Nacke et al 2011); 8, EM3L4 (Lee et al 2011); 9, FLS18 (Hu et al 2010b); 10, Est903 (Jia et al 2019); 11, EstJ (Choi et al 2013); 12, PE10 (Jiang et al 2012); 13, Est12 (Wu et al 2013); 14, EstDZ2 (Zarafeta et al 2016); 15, Est9x (Jeon et al 2009); 16, Lip10 (Guo et al 2016); 17, EstGH (Nacke et al 2011); 18, EML1 (Jeon et al 2009); 19, FnL (Yu et al 2010); 20, EstP2K (Ouyang et al 2013); 21, LipA (Couto et al 2010); 22, LipSM54 (Li et al 2016); 23, MtEst45 (Lee 2016); 24, LipT (Chow et al 2012); 25, EstSt7 (Wei et al 2013); 26, Rlip1 (Liu et al 2009); 27, EstA (Chu et al 2008); 28, FLS12 (Hu et al 2010b); 29, lp_3505 (Esteban-Torres et al 2014). Outer ring: substrate specificity of corresponding clones towards different carbon chain length (C4 – C14) of triglycerides.…”

Section: Resultsmentioning

confidence: 99%

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Metagenomic screening for lipolytic genes reveals an ecology-clustered distribution pattern

Schneider

Daniel

2021

Preprint

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Lipolytic enzymes are one of the most important enzyme types for application in various industrial processes. Despite the continuously increasing demand, only a small portion of the so far encountered lipolytic enzymes exhibit adequate stability and activities for biotechnological applications. To explore novel and/or extremophilic lipolytic enzymes, microbial consortia in two composts at thermophilic stage were analyzed using function-driven and sequence-based metagenomic approaches. Analysis of community composition by amplicon-based 16S rRNA genes and transcripts, and direct metagenome sequencing revealed that the communities of the compost samples were dominated by members of the phyla Actinobacteria, Proteobacteria, Firmicutes, Bacteroidetes and Chloroflexi. Function-driven screening of the metagenomic libraries constructed from the two samples yielded 115 unique genes encoding lipolytic enzymes. The family assignment of these enzymes was conducted by analyzing the phylogenetic relationship and generation of a protein sequence similarity network according to an integral classification system. The sequence-based screening was performed by using a newly developed database, containing a set of profile Hidden Markov models, highly sensitive and specific for detection of lipolytic enzymes. By comparing the lipolytic enzymes identified through both approaches, we demonstrated that the activity-directed complements sequence-based detection, and vice versa. The sequence-based comparative analysis of lipolytic genes regarding diversity, function and taxonomic origin derived from 175 metagenomes indicated significant differences between habitats. Analysis of the prevalent and distinct microbial groups providing the lipolytic genes revealed characteristic patterns and groups driven by ecological factors. The here presented data suggests that the diversity and distribution of lipolytic genes in metagenomes of various habitats are largely constrained by ecological factors.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Metagenomic screening for lipolytic genes reveals an ecology-clustered distribution pattern

Schneider

Daniel

2021

Preprint

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“…Carboxylesterases (EC 3.1.1.1, carboxylester hydrolases, ‘true’ esterase) which hydrolyze ester-containing molecules, and preferentially hydrolyze ‘simple’ esters and usually only triglycerides composed of fatty acids shorter than C6, and 2). Lipases (EC 3.1.1.3, triacylglycerol hydrolases, ‘true’ lipase) which catalyze the hydrolysis of water-insoluble substrates, such as triglycerides composed of long-chain fatty acids [ 14 , 15 , 16 ]. These enzymes are members of the α/β hydrolase superfamily [ 17 ].…”

Section: Introductionmentioning

confidence: 99%

“…Esterases can catalyze the hydrolysis of ester bonds or the synthesis of ester linkages [ 16 ]. In recent years, esterases that retain activity over a broad range of temperatures have been identified and used for industrial applications.…”

Section: Introductionmentioning

confidence: 99%

Identification and Biochemical Characterization of a Novel Hormone-Sensitive Lipase Family Esterase Est19 from the Antarctic Bacterium Pseudomonas sp. E2-15

et al. 2021

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Esterases represent an important class of enzymes with a wide variety of industrial applications. A novel hormone-sensitive lipase (HSL) family esterase, Est19, from the Antarctic bacterium Pseudomonas sp. E2-15 is identified, cloned, and expressed. The enzyme possesses a GESAG motif containing an active serine (S) located within a highly conserved catalytic triad of Ser155, Asp253, and His282 residues. The catalytic efficiency (kcat/Km) of Est19 for the pNPC6 substrate is 148.68 s−1mM−1 at 40 °C. Replacing Glu154 juxtaposed to the critical catalytic serine with Asp (E154→D substitution) reduced the activity and catalytic efficiency of the enzyme two-fold, with little change in the substrate affinity. The wild-type enzyme retained near complete activity over a temperature range of 10–60 °C, while ~50% of its activity was retained at 0 °C. A phylogenetic analysis suggested that Est19 and its homologs may represent a new subfamily of HSL. The thermal stability and stereo-specificity suggest that the Est19 esterase may be useful for cold and chiral catalyses.

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“…Arguably, for the classification of microorganisms in environmental samples, the metagenomic approach has gained prominence due to its potential for fast genetic analysis of all genomes contained in an environmental niche 6 . In addition, it is a technique that provides genetic information for the discovery of biocatalysts or novel enzymes from uncultivated organisms, providing important insights about a wide diversity of a bacterial community [7][8][9][10] . Relevant cell-cell communication is mediated by secretion of chemical compounds (autoinducer) to the environment, a process known as quorum-sensing (QS) 11,12 .…”

Section: Introductionmentioning

confidence: 99%

Molecular and functional basis of a novel Amazonian Dark Earth Esterase 1 (Ade1) with hysteresis behavior and quorum-quenching activity

Vinces

Souza

Carvalho

et al. 2020

Preprint

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Amazon Dark Earth (ADE) soil is rich in organic compounds and its fertility has been associated with a high diversity of microorganisms. Herein, we investigate the biochemical and functional features of a novel esterase, Ade1, obtained from a metagenomic library of Amazonian Dark Earth soils of the Amazonian Rainforest, in Brazil. The esterases cleave ester bonds to form a carboxylic and an alcohol group. Esterases and lipases are enzymes found in almost all living organisms, demonstrating their biological relevance. We reported that Ade1 belongs to an α/β-hydrolase superfamily. We suggest that Ade1 is a moonlighting enzyme with hysteresis behavior and quorum-quenching activity, which may play a key role in the metabolism of a Gram-negative proteobacteria. In addition, molecular dynamics simulations reveal that the hysteresis behavior is directly associated with structural properties of the cap domain. Our findings reveal details of the molecular basis, catalytic and structural mechanisms of a novel α/β-hydrolase, which may be applied to other esterases of biotechnological, food, and/or pharmaceutical interest.Abstract Figure

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Expression and characterization of an esterase belonging to a new family via isolation from a metagenomic library of paper mill sludge

Cited by 19 publications

References 33 publications

Metagenomic screening for lipolytic genes reveals an ecology-clustered distribution pattern

Metagenomic screening for lipolytic genes reveals an ecology-clustered distribution pattern

Identification and Biochemical Characterization of a Novel Hormone-Sensitive Lipase Family Esterase Est19 from the Antarctic Bacterium Pseudomonas sp. E2-15

Molecular and functional basis of a novel Amazonian Dark Earth Esterase 1 (Ade1) with hysteresis behavior and quorum-quenching activity

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