Enzyme catalysis with small ionic liquid quantities

Fischer, Fabian; Mutschler, Julien; Zufferey, Daniel

doi:10.1007/s10295-010-0908-1

Cited by 22 publications

(13 citation statements)

References 34 publications

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“…Furthermore, the subsequent hydrolysis of the biomass into fermentable sugars requires enzymes that remain active under conditions of high substrate loading and minimal solvent. The discovery of cellulose-dissolving ionic liquids in recent years suggests a new and 'greener' direction for processing of lignocellulosic materials [25-27] and to improve enzyme stability and activity under minimal solvent processing conditions [23]. However, there are concerns regarding retention of enzyme activities in the presence of ionic liquids [28].…”

Section: Resultsmentioning

confidence: 99%

“…Therefore, these four enzymes have the potential to be easily recovered after the treatment slurry and could be recycled. There is also the potential to use a His-tag to immobilize these enzymes and then apply them in a continuous reaction systems, eventually combined with the application of ionic liquids [23]. Further studies will be necessary to optimize conditions for specific reactions and perhaps improve the wild type enzyme performance.…”

Section: Discussionmentioning

confidence: 99%

“…The most active enzymes, identified by hydrolysis of chromophoric sugar aglycones, were selected for further investigation; including gene cloning, protein expression, and preliminary enzyme characterization. Activities of some enzymes were tested in the presence of ionic liquids, an emerging technology for biomass pretreatment as well as a new approach to increase enzyme stability and activity during minimal solvent processing [23]. …”

Section: Introductionmentioning

confidence: 99%

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Bioprospecting metagenomics of decaying wood: mining for new glycoside hydrolases

Taghavi

McCorkle

et al. 2011

Biotechnol Biofuels

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BackgroundTo efficiently deconstruct recalcitrant plant biomass to fermentable sugars in industrial processes, biocatalysts of higher performance and lower cost are required. The genetic diversity found in the metagenomes of natural microbial biomass decay communities may harbor such enzymes. Our goal was to discover and characterize new glycoside hydrolases (GHases) from microbial biomass decay communities, especially those from unknown or never previously cultivated microorganisms.ResultsFrom the metagenome sequences of an anaerobic microbial community actively decaying poplar biomass, we identified approximately 4,000 GHase homologs. Based on homology to GHase families/activities of interest and the quality of the sequences, candidates were selected for full-length cloning and subsequent expression. As an alternative strategy, a metagenome expression library was constructed and screened for GHase activities. These combined efforts resulted in the cloning of four novel GHases that could be successfully expressed in Escherichia coli. Further characterization showed that two enzymes showed significant activity on p-nitrophenyl-α-L-arabinofuranoside, one enzyme had significant activity against p-nitrophenyl-β-D-glucopyranoside, and one enzyme showed significant activity against p-nitrophenyl-β-D-xylopyranoside. Enzymes were also tested in the presence of ionic liquids.ConclusionsMetagenomics provides a good resource for mining novel biomass degrading enzymes and for screening of cellulolytic enzyme activities. The four GHases that were cloned may have potential application for deconstruction of biomass pretreated with ionic liquids, as they remain active in the presence of up to 20% ionic liquid (except for 1-ethyl-3-methylimidazolium diethyl phosphate). Alternatively, ionic liquids might be used to immobilize or stabilize these enzymes for minimal solvent processing of biomass.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Bioprospecting metagenomics of decaying wood: mining for new glycoside hydrolases

Taghavi

McCorkle

et al. 2011

Biotechnol Biofuels

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“…Using the his-tags, the enzymes, provided they are sufficiently stable, can be easily recovered from the treatment process and re-used in continuous applications, allowing alkaline and enhanced temperature conditions [53]. The here-detected activity of our enzymes (specifically proteins 1 and 2) under thermo-alkaliphilic conditions thus indicates their usefulness for plant biomass degradation, spurring the production of glucose and xylose/arabinose from (hemi)cellulosic material and also for pulp bleaching processes.…”

Section: Discussionmentioning

confidence: 99%

Molecular cloning, expression, and characterization of four novel thermo-alkaliphilic enzymes retrieved from a metagenomic library

Maruthamuthu

Elsas

2017

Biotechnol Biofuels

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BackgroundEnzyme discovery is a promising approach to aid in the deconstruction of recalcitrant plant biomass in an industrial process. Novel enzymes can be readily discovered by applying metagenomics on whole microbiomes. Our goal was to select, examine, and characterize eight novel glycoside hydrolases that were previously detected in metagenomic libraries, to serve biotechnological applications with high performance.ResultsHere, eight glycosyl hydrolase family candidate genes were selected from metagenomes of wheat straw-degrading microbial consortia using molecular cloning and subsequent gene expression studies in Escherichia coli. Four of the eight enzymes had significant activities on either pNP-β-d-galactopyranoside, pNP-β-d-xylopyranoside, pNP-α-l-arabinopyranoside or pNP-α-d-glucopyranoside. These proteins, denoted as proteins 1, 2, 5 and 6, were his-tag purified and their nature and activities further characterized using molecular and activity screens with the pNP-labeled substrates. Proteins 1 and 2 showed high homologies with (1) a β-galactosidase (74%) and (2) a β-xylosidase (84%), whereas the remaining two (5 and 6) were homologous with proteins reported as a diguanylate cyclase and an aquaporin, respectively. The β-galactosidase- and β-xylosidase-like proteins 1 and 2 were confirmed as being responsible for previously found thermo-alkaliphilic glycosidase activities of extracts of E. coli carrying the respective source fosmids. Remarkably, the β-xylosidase-like protein 2 showed activities with both pNP-Xyl and pNP-Ara in the temperature range 40–50 °C and pH range 8.0–10.0. Moreover, proteins 5 and 6 showed thermotolerant α-glucosidase activity at pH 10.0. In silico structure prediction of protein 5 revealed the presence of a potential “GGDEF” catalytic site, encoding α-glucosidase activity, whereas that of protein 6 showed a “GDSL” site, encoding a ‘new family’ α-glucosidase activity.ConclusionUsing a rational screening approach, we identified and characterized four thermo-alkaliphilic glycosyl hydrolases that have the potential to serve as constituents of enzyme cocktails that produce sugars from lignocellulosic plant remains.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-017-0808-y) contains supplementary material, which is available to authorized users.

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“…This indicates that the enzymatic activities and hydrolysis yield of polydatin to trans-resveratrol increased from 0 to 0.5 M and then decreased from 0.5 to 1.0 M. This may be explained as follows: the extraction and hydrolysis results for polydatin and trans-resveratrol represent the combined effects of the ionic-liquid solution and the enzyme catalytic conversion function. According to the literature[45,46], ionic liquids have a certain inhibitory effect on activities of enzymes. Low ionic-liquid concentration could improve reaction rates, but the greater the ionic-liquid concentration, the stronger the inhibition[36].…”

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confidence: 99%

A new approach for obtaining trans-resveratrol from tree peony seed oil extracted residues using ionic liquid-based enzymatic hydrolysis in situ extraction

Chen

Zhang²,

et al. 2016

Separation and Purification Technology

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Enzyme catalysis with small ionic liquid quantities

Cited by 22 publications

References 34 publications

Bioprospecting metagenomics of decaying wood: mining for new glycoside hydrolases

Bioprospecting metagenomics of decaying wood: mining for new glycoside hydrolases

Molecular cloning, expression, and characterization of four novel thermo-alkaliphilic enzymes retrieved from a metagenomic library

A new approach for obtaining trans-resveratrol from tree peony seed oil extracted residues using ionic liquid-based enzymatic hydrolysis in situ extraction

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