High-Throughput Screening Assay for Laccase Engineering toward Lignosulfonate Valorization

Rodriguez-Escribano, David; de Salas, Felipe; Pardo, Isabel; Camarero, Susana

doi:10.3390/ijms18081793

Title: High-Throughput Screening Assay for Laccase Engineering toward Lignosulfonate Valorization

Abstract

Lignin valorization is a pending issue for the integrated conversion of lignocellulose in consumer goods. Lignosulfonates (LS) are the main technical lignins commercialized today. However, their molecular weight should be enlarged to meet application requirements as additives or dispersing agents. Oxidation of lignosulfonates with fungal oxidoreductases, such as laccases, can increase the molecular weight of lignosulfonates by the cross-linking of lignin phenols. To advance in this direction, we describe here the development of a high-throughput screening (HTS) assay for the directed evolution of laccases, with lignosulfonate as substrate and the Folin-Ciocalteau reagent (FCR), to detect the decrease in phenolic content produced upon polymerization of lignosulfonate by the enzyme. Once the reaction conditions were adjusted to the 96-well-plate format, the enzyme for validating the assay was selected from a battery of high-redox-potential laccase variants functionally expressed in S. cerevisiae (the preferred host for the directed evolution of fungal oxidoreductases). The colorimetric response (absorbance at 760 nm) correlated with laccase activity secreted by the yeast. The HTS assay was reproducible (coefficient of variation (CV) = 15%) and sensitive enough to detect subtle differences in activity among yeast clones expressing a laccase mutant library obtained by error-prone PCR (epPCR). As a result, themore »« less

Authors:

Rodriguez-Escribano, David ^[1]; de Salas, Felipe ^[1]; Pardo, Isabel ^[2];

^[1]

Centro de Investigaciones Biologicas, Madrid (Spain)
Centro de Investigaciones Biologicas, Madrid (Spain); National Renewable Energy Lab. (NREL), Golden, CO (United States)

Publication Date:: Fri Aug 18 00:00:00 EDT 2017

Research Org.:: National Renewable Energy Laboratory (NREL), Golden, CO (United States)

Sponsoring Org.:: USDOE Office of Energy Efficiency and Renewable Energy (EERE)

OSTI Identifier:: 1378888

Report Number(s):: NREL/JA-5100-69122
Journal ID: ISSN 1422-0067; IJMCFK

Grant/Contract Number:: AC36-08GO28308

Resource Type:: Accepted Manuscript

Journal Name:: International Journal of Molecular Sciences (Online)

Additional Journal Information:: Journal Name: International Journal of Molecular Sciences (Online); Journal Volume: 18; Journal Issue: 8; Journal ID: ISSN 1422-0067

Publisher:: MDPI

Country of Publication:: United States

Language:: English

Subject:: 09 BIOMASS FUELS; laccase; lignosulfonate; high-throughput screening; phenolic content; enzyme directed evolution

Citation Formats


                    Rodriguez-Escribano, David, de Salas, Felipe, Pardo, Isabel, and Camarero, Susana. High-Throughput Screening Assay for Laccase Engineering toward Lignosulfonate Valorization.  United States: N. p., 2017. 
Web.  doi:10.3390/ijms18081793.

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                    Rodriguez-Escribano, David, de Salas, Felipe, Pardo, Isabel, & Camarero, Susana. High-Throughput Screening Assay for Laccase Engineering toward Lignosulfonate Valorization.  United States.  https://doi.org/10.3390/ijms18081793

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                    Rodriguez-Escribano, David, de Salas, Felipe, Pardo, Isabel, and Camarero, Susana. Fri .  
"High-Throughput Screening Assay for Laccase Engineering toward Lignosulfonate Valorization".  United States.  https://doi.org/10.3390/ijms18081793.  https://www.osti.gov/servlets/purl/1378888.

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@article{osti_1378888,

  title        = {High-Throughput Screening Assay for Laccase Engineering toward Lignosulfonate Valorization},

  author       = {Rodriguez-Escribano, David and de Salas, Felipe and Pardo, Isabel and Camarero, Susana},

  abstractNote = {Lignin valorization is a pending issue for the integrated conversion of lignocellulose in consumer goods. Lignosulfonates (LS) are the main technical lignins commercialized today. However, their molecular weight should be enlarged to meet application requirements as additives or dispersing agents. Oxidation of lignosulfonates with fungal oxidoreductases, such as laccases, can increase the molecular weight of lignosulfonates by the cross-linking of lignin phenols. To advance in this direction, we describe here the development of a high-throughput screening (HTS) assay for the directed evolution of laccases, with lignosulfonate as substrate and the Folin-Ciocalteau reagent (FCR), to detect the decrease in phenolic content produced upon polymerization of lignosulfonate by the enzyme. Once the reaction conditions were adjusted to the 96-well-plate format, the enzyme for validating the assay was selected from a battery of high-redox-potential laccase variants functionally expressed in S. cerevisiae (the preferred host for the directed evolution of fungal oxidoreductases). The colorimetric response (absorbance at 760 nm) correlated with laccase activity secreted by the yeast. The HTS assay was reproducible (coefficient of variation (CV) = 15%) and sensitive enough to detect subtle differences in activity among yeast clones expressing a laccase mutant library obtained by error-prone PCR (epPCR). As a result, the method is therefore feasible for screening thousands of clones during the precise engineering of laccases toward valorization of lignosulfonates.},

  doi          = {10.3390/ijms18081793},

  journal      = {International Journal of Molecular Sciences (Online)},

  number       = 8,

  volume       = 18,

  place        = {United States},

  year         = {Fri Aug 18 00:00:00 EDT 2017},

  month        = {Fri Aug 18 00:00:00 EDT 2017}

}

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Works referenced in this record:

Evaluating Lignins as Enzyme Substrates: Insights and Methodological Recommendations from a Study of Laccase-Catalyzed Lignin Polymerization
journal, February 2014

West, Mark A.; Hickson, Aynsley C.; Mattinen, Maija-Liisa
BioResources, Vol. 9, Issue 2
DOI: 10.15376/biores.9.2.2782-2796

Degradation mechanisms of phenolic β-1 lignin substructure model compounds by laccase of Coriolus versicolor
journal, April 1988

Kawai, Shingo; Umezawa, Toshiaki; Higuchi, Takayoshi
Archives of Biochemistry and Biophysics, Vol. 262, Issue 1
DOI: 10.1016/0003-9861(88)90172-5

Exploring the Oxidation of Lignin-Derived Phenols by a Library of Laccase Mutants
journal, September 2015

Pardo, Isabel; Camarero, Susana
Molecules, Vol. 20, Issue 9
DOI: 10.3390/molecules200915929

Laccase/mediated oxidation of a lignin model for improved delignification procedures
journal, November 2003

Barreca, Anna Maria; Fabbrini, Maura; Galli, Carlo
Journal of Molecular Catalysis B: Enzymatic, Vol. 26, Issue 1-2
DOI: 10.1016/j.molcatb.2003.08.001

Investigation of the Molecular Weight Increase of Commercial Lignosulfonates by Laccase Catalysis
journal, April 2010

Areskogh, Dimitri; Li, Jiebing; Gellerstedt, Göran
Biomacromolecules, Vol. 11, Issue 4
DOI: 10.1021/bm901258v

Structure and Properties of Sodium Lignosulfonate with Different Molecular Weight Used as Dye Dispersant
journal, November 2014

Yang, Dongjie; Li, Huijing; Qin, Yanlin
Journal of Dispersion Science and Technology, Vol. 36, Issue 4
DOI: 10.1080/01932691.2014.916221

Characterisation of enzymatically oxidised lignosulfonates and their application on lignocellulosic fabrics
journal, August 2009

Kim, Suyeon; Silva, Carla; Zille, Andrea
Polymer International, Vol. 58, Issue 8
DOI: 10.1002/pi.2600

Engineering Platforms for Directed Evolution of Laccase from Pycnoporus cinnabarinus
journal, December 2011

Camarero, S.; Pardo, I.; Cañas, A. I.
Applied and Environmental Microbiology, Vol. 78, Issue 5
DOI: 10.1128/AEM.07530-11

Horseradish Peroxidase Modification of Sulfomethylated Wheat Straw Alkali Lignin To Improve Its Dispersion Performance
journal, February 2015

Zhou, Haifeng; Chang, Yaqi; Wu, Xiaolei
ACS Sustainable Chemistry & Engineering, Vol. 3, Issue 3
DOI: 10.1021/sc500757y

Enzymatic polymerization of sodium lignosulfonates: effect of catalysts, initial molecular weight, and mediators
journal, March 2013

Madad, Nidal; Chebil, Latifa; Charbonnel, Celine
Canadian Journal of Chemistry, Vol. 91, Issue 3
DOI: 10.1139/cjc-2012-0036

Activation of Magnesium Lignosulfonate and Kraft Lignin: Influence on the Properties of Phenolic Resin-Based Composites for Potential Applications in Abrasive Materials
journal, June 2017

Klapiszewski, Lukasz; Jamrozik, Artur; Strzemiecka, Beata
International Journal of Molecular Sciences, Vol. 18, Issue 6
DOI: 10.3390/ijms18061224

Computer-Aided Laccase Engineering: Toward Biological Oxidation of Arylamines
journal, July 2016

Santiago, Gerard; de Salas, Felipe; Lucas, M. Fátima
ACS Catalysis, Vol. 6, Issue 8
DOI: 10.1021/acscatal.6b01460

Polymerization of Various Lignins via Immobilized Myceliophthora thermophila Laccase (MtL)
journal, August 2016

Huber, Daniela; Pellis, Alessandro; Daxbacher, Andreas
Polymers, Vol. 8, Issue 8
DOI: 10.3390/polym8080280

Development of chimeric laccases by directed evolution
journal, July 2012

Pardo, Isabel; Vicente, Ana Isabel; Mate, Diana M.
Biotechnology and Bioengineering, Vol. 109, Issue 12
DOI: 10.1002/bit.24588

Polymerization of lignosulfonates by the laccase-HBT (1-hydroxybenzotriazole) system improves dispersibility
journal, July 2010

Nugroho Prasetyo, Endry; Kudanga, Tukayi; Østergaard, Lars
Bioresource Technology, Vol. 101, Issue 14
DOI: 10.1016/j.biortech.2010.01.048

Influence of Oxygen and Mediators on Laccase-Catalyzed Polymerization of Lignosulfonate
journal, June 2016

Huber, Daniela; Ortner, Andreas; Daxbacher, Andreas
ACS Sustainable Chemistry & Engineering, Vol. 4, Issue 10
DOI: 10.1021/acssuschemeng.6b00692

Demonstration of lignin-to-peroxidase direct electron transfer. A TRANSIENT-STATE KINETICS, DIRECTED MUTAGENESIS, EPR AND NMR STUDY.
journal, December 2015

Sáez-Jiménez, Verónica; Baratto, Maria Camilla; Pogni, Rebecca
Journal of Biological Chemistry, Vol. 290, Issue 51
DOI: 10.1074/jbc.a115.665919

Development of chimeric laccases by directed evolution
journal, July 2012

Pardo, Isabel; Vicente, Ana Isabel; Mate, Diana M.
Biotechnology and Bioengineering, Vol. 109, Issue 12
DOI: 10.1002/bit.24588

Characterisation of enzymatically oxidised lignosulfonates and their application on lignocellulosic fabrics
journal, August 2009

Kim, Suyeon; Silva, Carla; Zille, Andrea
Polymer International, Vol. 58, Issue 8
DOI: 10.1002/pi.2600

Degradation mechanisms of phenolic β-1 lignin substructure model compounds by laccase of Coriolus versicolor
journal, April 1988

Kawai, Shingo; Umezawa, Toshiaki; Higuchi, Takayoshi
Archives of Biochemistry and Biophysics, Vol. 262, Issue 1
DOI: 10.1016/0003-9861(88)90172-5

Polymerization of lignosulfonates by the laccase-HBT (1-hydroxybenzotriazole) system improves dispersibility
journal, July 2010

Nugroho Prasetyo, Endry; Kudanga, Tukayi; Østergaard, Lars
Bioresource Technology, Vol. 101, Issue 14
DOI: 10.1016/j.biortech.2010.01.048

Computer-Aided Laccase Engineering: Toward Biological Oxidation of Arylamines
journal, July 2016

Santiago, Gerard; de Salas, Felipe; Lucas, M. Fátima
ACS Catalysis, Vol. 6, Issue 8
DOI: 10.1021/acscatal.6b01460

Influence of Oxygen and Mediators on Laccase-Catalyzed Polymerization of Lignosulfonate
journal, June 2016

Huber, Daniela; Ortner, Andreas; Daxbacher, Andreas
ACS Sustainable Chemistry & Engineering, Vol. 4, Issue 10
DOI: 10.1021/acssuschemeng.6b00692

Investigation of the Molecular Weight Increase of Commercial Lignosulfonates by Laccase Catalysis
journal, April 2010

Areskogh, Dimitri; Li, Jiebing; Gellerstedt, Göran
Biomacromolecules, Vol. 11, Issue 4
DOI: 10.1021/bm901258v

Horseradish Peroxidase Modification of Sulfomethylated Wheat Straw Alkali Lignin To Improve Its Dispersion Performance
journal, February 2015

Zhou, Haifeng; Chang, Yaqi; Wu, Xiaolei
ACS Sustainable Chemistry & Engineering, Vol. 3, Issue 3
DOI: 10.1021/sc500757y

The pros and cons of lignin valorisation in an integrated biorefinery
journal, January 2014

Strassberger, Zea; Tanase, Stefania; Rothenberg, Gadi
RSC Adv., Vol. 4, Issue 48
DOI: 10.1039/c4ra04747h

Demonstration of Lignin-to-Peroxidase Direct Electron Transfer: A TRANSIENT-STATE KINETICS, DIRECTED MUTAGENESIS, EPR, AND NMR STUDY
journal, August 2015

Sáez-Jiménez, Verónica; Baratto, Maria Camilla; Pogni, Rebecca
Journal of Biological Chemistry, Vol. 290, Issue 38
DOI: 10.1074/jbc.m115.665919

Lignin oxidation by laccase isozymes from Trametes versicolor and role of the mediator 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonate) in kraft lignin depolymerization.
journal, January 1995

Bourbonnais, R.; Paice, M. G.; Reid, I. D.
Applied and environmental microbiology, Vol. 61, Issue 5
DOI: 10.1128/aem.61.5.1876-1880.1995

Evaluating Lignins as Enzyme Substrates: Insights and Methodological Recommendations from a Study of Laccase-Catalyzed Lignin Polymerization
journal, February 2014

West, Mark A.; Hickson, Aynsley C.; Mattinen, Maija-Liisa
BioResources, Vol. 9, Issue 2
DOI: 10.15376/biores.9.2.2782-2796

Activation of Magnesium Lignosulfonate and Kraft Lignin: Influence on the Properties of Phenolic Resin-Based Composites for Potential Applications in Abrasive Materials
journal, June 2017

Klapiszewski, Lukasz; Jamrozik, Artur; Strzemiecka, Beata
International Journal of Molecular Sciences, Vol. 18, Issue 6
DOI: 10.3390/ijms18061224

Exploring the Oxidation of Lignin-Derived Phenols by a Library of Laccase Mutants
journal, September 2015

Pardo, Isabel; Camarero, Susana
Molecules, Vol. 20, Issue 9
DOI: 10.3390/molecules200915929

Works referencing / citing this record:

A highly stable laccase obtained by swapping the second cupredoxin domain
journal, October 2018

Pardo, Isabel; Rodríguez-Escribano, David; Aza, Pablo
Scientific Reports, Vol. 8, Issue 1
DOI: 10.1038/s41598-018-34008-3

Alternatives for Chemical and Biochemical Lignin Valorization: Hot Topics from a Bibliometric Analysis of the Research Published During the 2000–2016 Period
journal, July 2018

Abejón, Ricardo; Pérez-Acebo, Heriberto; Clavijo, Leonardo
Processes, Vol. 6, Issue 8
DOI: 10.3390/pr6080098

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Similar Records

Title: High-Throughput Screening Assay for Laccase Engineering toward Lignosulfonate Valorization

Abstract

Citation Formats

Evaluating Lignins as Enzyme Substrates: Insights and Methodological Recommendations from a Study of Laccase-Catalyzed Lignin Polymerization journal, February 2014

Degradation mechanisms of phenolic β-1 lignin substructure model compounds by laccase of Coriolus versicolor journal, April 1988

Exploring the Oxidation of Lignin-Derived Phenols by a Library of Laccase Mutants journal, September 2015

Laccase/mediated oxidation of a lignin model for improved delignification procedures journal, November 2003

Investigation of the Molecular Weight Increase of Commercial Lignosulfonates by Laccase Catalysis journal, April 2010

Structure and Properties of Sodium Lignosulfonate with Different Molecular Weight Used as Dye Dispersant journal, November 2014

Characterisation of enzymatically oxidised lignosulfonates and their application on lignocellulosic fabrics journal, August 2009

Engineering Platforms for Directed Evolution of Laccase from Pycnoporus cinnabarinus journal, December 2011

Horseradish Peroxidase Modification of Sulfomethylated Wheat Straw Alkali Lignin To Improve Its Dispersion Performance journal, February 2015

Enzymatic polymerization of sodium lignosulfonates: effect of catalysts, initial molecular weight, and mediators journal, March 2013

Activation of Magnesium Lignosulfonate and Kraft Lignin: Influence on the Properties of Phenolic Resin-Based Composites for Potential Applications in Abrasive Materials journal, June 2017

Computer-Aided Laccase Engineering: Toward Biological Oxidation of Arylamines journal, July 2016

Polymerization of Various Lignins via Immobilized Myceliophthora thermophila Laccase (MtL) journal, August 2016

Development of chimeric laccases by directed evolution journal, July 2012

Polymerization of lignosulfonates by the laccase-HBT (1-hydroxybenzotriazole) system improves dispersibility journal, July 2010

Influence of Oxygen and Mediators on Laccase-Catalyzed Polymerization of Lignosulfonate journal, June 2016

Demonstration of lignin-to-peroxidase direct electron transfer. A TRANSIENT-STATE KINETICS, DIRECTED MUTAGENESIS, EPR AND NMR STUDY. journal, December 2015

Development of chimeric laccases by directed evolution journal, July 2012

Characterisation of enzymatically oxidised lignosulfonates and their application on lignocellulosic fabrics journal, August 2009

Degradation mechanisms of phenolic β-1 lignin substructure model compounds by laccase of Coriolus versicolor journal, April 1988

Polymerization of lignosulfonates by the laccase-HBT (1-hydroxybenzotriazole) system improves dispersibility journal, July 2010

Computer-Aided Laccase Engineering: Toward Biological Oxidation of Arylamines journal, July 2016

Influence of Oxygen and Mediators on Laccase-Catalyzed Polymerization of Lignosulfonate journal, June 2016

Investigation of the Molecular Weight Increase of Commercial Lignosulfonates by Laccase Catalysis journal, April 2010

Horseradish Peroxidase Modification of Sulfomethylated Wheat Straw Alkali Lignin To Improve Its Dispersion Performance journal, February 2015

The pros and cons of lignin valorisation in an integrated biorefinery journal, January 2014

Demonstration of Lignin-to-Peroxidase Direct Electron Transfer: A TRANSIENT-STATE KINETICS, DIRECTED MUTAGENESIS, EPR, AND NMR STUDY journal, August 2015

Lignin oxidation by laccase isozymes from Trametes versicolor and role of the mediator 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonate) in kraft lignin depolymerization. journal, January 1995

Evaluating Lignins as Enzyme Substrates: Insights and Methodological Recommendations from a Study of Laccase-Catalyzed Lignin Polymerization journal, February 2014

Activation of Magnesium Lignosulfonate and Kraft Lignin: Influence on the Properties of Phenolic Resin-Based Composites for Potential Applications in Abrasive Materials journal, June 2017

Exploring the Oxidation of Lignin-Derived Phenols by a Library of Laccase Mutants journal, September 2015

A highly stable laccase obtained by swapping the second cupredoxin domain journal, October 2018

Alternatives for Chemical and Biochemical Lignin Valorization: Hot Topics from a Bibliometric Analysis of the Research Published During the 2000–2016 Period journal, July 2018

Evaluating Lignins as Enzyme Substrates: Insights and Methodological Recommendations from a Study of Laccase-Catalyzed Lignin Polymerization
journal, February 2014

Degradation mechanisms of phenolic β-1 lignin substructure model compounds by laccase of Coriolus versicolor
journal, April 1988

Exploring the Oxidation of Lignin-Derived Phenols by a Library of Laccase Mutants
journal, September 2015

Laccase/mediated oxidation of a lignin model for improved delignification procedures
journal, November 2003

Investigation of the Molecular Weight Increase of Commercial Lignosulfonates by Laccase Catalysis
journal, April 2010

Structure and Properties of Sodium Lignosulfonate with Different Molecular Weight Used as Dye Dispersant
journal, November 2014

Characterisation of enzymatically oxidised lignosulfonates and their application on lignocellulosic fabrics
journal, August 2009

Engineering Platforms for Directed Evolution of Laccase from Pycnoporus cinnabarinus
journal, December 2011

Horseradish Peroxidase Modification of Sulfomethylated Wheat Straw Alkali Lignin To Improve Its Dispersion Performance
journal, February 2015

Enzymatic polymerization of sodium lignosulfonates: effect of catalysts, initial molecular weight, and mediators
journal, March 2013

Activation of Magnesium Lignosulfonate and Kraft Lignin: Influence on the Properties of Phenolic Resin-Based Composites for Potential Applications in Abrasive Materials
journal, June 2017

Computer-Aided Laccase Engineering: Toward Biological Oxidation of Arylamines
journal, July 2016

Polymerization of Various Lignins via Immobilized Myceliophthora thermophila Laccase (MtL)
journal, August 2016

Development of chimeric laccases by directed evolution
journal, July 2012

Polymerization of lignosulfonates by the laccase-HBT (1-hydroxybenzotriazole) system improves dispersibility
journal, July 2010

Influence of Oxygen and Mediators on Laccase-Catalyzed Polymerization of Lignosulfonate
journal, June 2016

Demonstration of lignin-to-peroxidase direct electron transfer. A TRANSIENT-STATE KINETICS, DIRECTED MUTAGENESIS, EPR AND NMR STUDY.
journal, December 2015

Development of chimeric laccases by directed evolution
journal, July 2012

Characterisation of enzymatically oxidised lignosulfonates and their application on lignocellulosic fabrics
journal, August 2009

Degradation mechanisms of phenolic β-1 lignin substructure model compounds by laccase of Coriolus versicolor
journal, April 1988

Polymerization of lignosulfonates by the laccase-HBT (1-hydroxybenzotriazole) system improves dispersibility
journal, July 2010

Computer-Aided Laccase Engineering: Toward Biological Oxidation of Arylamines
journal, July 2016

Influence of Oxygen and Mediators on Laccase-Catalyzed Polymerization of Lignosulfonate
journal, June 2016

Investigation of the Molecular Weight Increase of Commercial Lignosulfonates by Laccase Catalysis
journal, April 2010

Horseradish Peroxidase Modification of Sulfomethylated Wheat Straw Alkali Lignin To Improve Its Dispersion Performance
journal, February 2015

The pros and cons of lignin valorisation in an integrated biorefinery
journal, January 2014

Demonstration of Lignin-to-Peroxidase Direct Electron Transfer: A TRANSIENT-STATE KINETICS, DIRECTED MUTAGENESIS, EPR, AND NMR STUDY
journal, August 2015

Lignin oxidation by laccase isozymes from Trametes versicolor and role of the mediator 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonate) in kraft lignin depolymerization.
journal, January 1995

Evaluating Lignins as Enzyme Substrates: Insights and Methodological Recommendations from a Study of Laccase-Catalyzed Lignin Polymerization
journal, February 2014

Activation of Magnesium Lignosulfonate and Kraft Lignin: Influence on the Properties of Phenolic Resin-Based Composites for Potential Applications in Abrasive Materials
journal, June 2017

Exploring the Oxidation of Lignin-Derived Phenols by a Library of Laccase Mutants
journal, September 2015

A highly stable laccase obtained by swapping the second cupredoxin domain
journal, October 2018

Alternatives for Chemical and Biochemical Lignin Valorization: Hot Topics from a Bibliometric Analysis of the Research Published During the 2000–2016 Period
journal, July 2018