Effects of organic solvents on the activity of free and immobilised laccase from Rhus vernicifera

Wan, Youchuan; Lü, Rong; Xiao, Ling; Du, Yumin; Miyakoshi, Tetsuo; Chen, Chen-Loung; Knill, Charles J.; Kennedy, John F.

doi:10.1016/j.ijbiomac.2010.07.003

Cited by 59 publications

(30 citation statements)

References 49 publications

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“…Recent example of Rhus vernicifera laccase losing more than 85% of initial activity in 20% ethanol and methanol may further emphasize significance of BliLacc resistance to organic solvents inactivation [33].…”

Section: Discussionmentioning

confidence: 99%

Expression and characterization of a thermostable organic solvent-tolerant laccase from Bacillus licheniformis ATCC 9945a

Lončar

Božić

Vujčić

2016

Journal of Molecular Catalysis B: Enzymatic

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Please cite this article as: Nikola Lončar, Nataša Božić, Zoran Vujčić, Expression and characterization of a thermostable organic solevent-tolerant laccase from Bacillus licheniformis ATCC 9945a, Journal of Molecular Catalysis B: Enzymatic http://dx.Abstract Highlights  Bacillus licheniformis 9945a laccase is overexpressed in E. coli with yield 50 mg/L.  Temperature optimum of laccase is 90⁰C and pH optimum is 7.0. Enzyme is thermostable with a melting temperature of 79⁰C at pH 7.0. Presence of organic solvents reduces melting temperature but activity remains impaired. Lignin model compounds are dimerized after one electron oxidation of phenolic group. AbstractBacterial laccases have proven advantages over fungal and plant counterparts in terms of wider pH optimum, higher stability and broader biocatalytic scope. In this work, Bacillus licheniformis ATCC 9945a laccase is produced heterologously in Escherichia coli. Produced laccase exhibits remarkably high temperature optimum at 90⁰C and possess significant thermostability and resistance to inactivation by organic solvents. Laccase has an apparent melting temperature of 79⁰C at pH 7.0 and above 70⁰C in range of pH 5.0-8.0, while having half-life of 50 min at 70⁰C. Presence of 10% organic solvents such as acetonitrile, dimethylformamide, dimethylsulfoxide or methanol reduces melting temperature to 45-52ᴼC but activity remains practically unimpaired. With 50% of acetonitrile and methanol laccase retained 3 ~40% of initial activity. EDTA and 300 mM sodium-chloride have positive effect on activity.Enzyme is active on syringaldazine, ABTS, phenols, amines, naphthol, lignin and lignin model compounds and mediates C-C bond formation via oxidative coupling after one electron oxidation of phenolic group. Successful polymerization of 2-naphthol was achieved with 77% conversion of 250 mg/L 2-naphtol in only 15 min which may further expand substrate scope of this enzyme towards polymer production and/or xenobiotics removal for environmental applications.

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Section: Discussionmentioning

confidence: 99%

Expression and characterization of a thermostable organic solvent-tolerant laccase from Bacillus licheniformis ATCC 9945a

Lončar

Božić

Vujčić

2016

Journal of Molecular Catalysis B: Enzymatic

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“…1.10.3.2 para -diphenol: oxygen oxidoreductase) is a cuproprotein that can catalyze the one-electron oxidation of a wide array of substrates with the simultaneous reduction of oxygen to water (D ' Annibale 1999, D ' Annibale 2000, Jiang et al 2005. Laccases can be produced by fungi, higher plants, and some bacteria (Dur á n et al 2002, Jolivalta et al 2000, Wan et al 2010, Rotkova et al 2009). …”

Section: Introductionmentioning

confidence: 99%

Immobilization of laccase on polyacrylamide and polyacrylamide – κ – carragennan-based semi-interpenetrating polymer networks

Gökgöz

Altinok

2012

Artificial Cells, Blood Substitutes, and Biotechnology

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In this study, laccase enzyme (L) from Agaricus bisporus was immobilized by entrapment into polyacrylamide (PAAm) and semi-interpenetrating polymer networks (semi-IPNs) prepared with either polyacrylamide/κ-carragennan (0.05g) [PAAm/ κ-car (0.05)] or polyacrylamide/κ-carragennan (0.1 g) [PAAm/ κ-car (0.1)]. The optimum pH was 6.0 for free L, 8.0 for PAAm-L, 8.5 for PAAm/κ-car (0.05)-L, and 9.0 for PAAm/κ-car (0.1)-L. The optimum temperature was determined as 45°C for free L and 60°C for all immobilized laccases. After 27 days of storage at 4°C, free enzyme lost its initial activity whereas immobilized enzymes retained 56 % (-)80% of their initial activities. The immobilized samples were used repeatedly 35 times by retaining 28 %-58 % of their initial activity. K(m(app)) values were calculated as 0.088, 0.139, 0.133, and 0.131 mM and Vmax values were found to be 2.83 x 10(-3), 4.51×10(-3), 4.76×10(-3), and 4.97×10(-3) mM min(-1) for free L and PAAm-L, PAAm/κ-car (0.05)-L, and PAAm/κ-car (0.1)-L, respectively.

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“…The use of water miscible solvents still required a water content of > 65% to maintain enzyme activity. Variations in the activity of the immobilized enzyme in the three solvents tested can be attributed to differences in the solvation layer [9] of the substrate and inhibitor and the products of the enzymatic reaction and the different partition between the solvent media and the microenvironment of the immobilized enzyme.…”

Section: Paraoxon Detection In Organic Solventsmentioning

confidence: 99%

“…Enzyme stability and activity are dependent on the local environment and can vary with the hydrophilicity/hydrophobicity of the solvent [8,9]. Enzymes in organic solvents often display reduced activity and selectivity as compared to water due to reduced conformational flexibility, altered substrate/inhibitor partition, removal of the hydration shell, and pH change in the enzyme microenvironment [6,8,10,11].…”

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confidence: 99%

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An acetylcholinesterase (AChE) biosensor with enhanced solvent resistance based on chitosan for the detection of pesticides

Warner

Andreescu

2016

Talanta

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Effects of organic solvents on the activity of free and immobilised laccase from Rhus vernicifera

Cited by 59 publications

References 49 publications

Expression and characterization of a thermostable organic solvent-tolerant laccase from Bacillus licheniformis ATCC 9945a

Expression and characterization of a thermostable organic solvent-tolerant laccase from Bacillus licheniformis ATCC 9945a

Immobilization of laccase on polyacrylamide and polyacrylamide – κ – carragennan-based semi-interpenetrating polymer networks

An acetylcholinesterase (AChE) biosensor with enhanced solvent resistance based on chitosan for the detection of pesticides

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