CBMA

Publications (25/10/17): 44; Granted International Patents: 2; Citations: 4118; H-index: 19
Principal investigator and inventor in the successful development and worldwide marketing of a xylanase for use in the food industry, currently being marketed worldwide by Puratos N.V.

 My domain of specialisation may be said to be in white (industrial) biotechnology and in the study and development of proteins for use in this. The principal focus is on protein based polymers (PBPs) and cold-adapted enzymes, their fundamentals and application in industry.

I have a strong background in the study of proteins; their production and purification, physiochemical and biophysical characterisation and in protein structure determination and protein engineering. My focus has mainly been on glycoside hydrolases (in particular xylanases) and oxidoreductases with both fundamental and applied aspects being covered. Recently attention has been turned towards protein based polymers (silk-elastin based) and approaches for the large scale production and purification of these. In addition, I have experience in the valorisation of scientific research, with the successful commercialisation of enzymes for use in the food industry and a current involvement in the development of protein based polymers for the automobile industry.

Research Areas

1) Extremophilic Enzymes: adaptation to temperature and biotechnological applications of extremophiles.
Keywords: protein science/enzymology/enzyme kinetics/protein structure (X-ray and NMR) and function/protein dynamics/protein engineering (site directed and random mutagenesis)/biophysics/application of enzymes in industry/glycoside hydrolases/cold-adapted enzymes/food and feed industries.

2) ECOplast: Research in new biomass-based composites from renewable resources with improved properties for vehicle parts moulding.
Development of industrial scale production and purification procedures for protein based polymers.
Keywords: protein based polymer/molecular biology/recombinant protein production/batch and fed-batch production in E. coli/pilot and industrial scale recombinant protein production/laboratory and industrial scale protein purification/protein based polymer characterisation.
 
1) Extremophilic Enzymes: The researcher has a strong background and experience in the study of enzymes produced by organisms inhabiting extreme environments and in particular in understanding the adaptation strategies used by cold-adapted enzymes in allowing them to function efficiently at low temperatures. The principal focus of the researcher has been on glycoside hydrolases (and in particular xylanases), laccases and oxidoreductases. The work has involved studies in comparative protein science: molecular biology, physicochemical studies, enzyme kinetics, protein structure, function and dynamics and biophysical studies as well as investigation of the application of cold adapted enzymes in the food industry. Indeed the researcher is internationally recognised in the field of cold-adapted enzymes and xylanases with numerous invitations for highly cited (approx. 2600 citations) papers and reviews (e.g. FEMS Microbiol. Rev., EMBO Rep., book chapters etc.) as well as key note lectures at International scientific conferences (e.g. Gordon Research Conference, Ventura, USA; Plant and Microbe Adaptations to Cold Meeting, Salsomaggiore Terme, Italy). Furthermore, this work has resulted in 2 granted international patents (worldwide) for which the present researcher is the principal inventor (52%) and the successful development of a cold-adapted xylanase for use in baking as well as its worldwide commercialisation by an industrial partner (Puratos N.V.).
    
2) ECOplast: This is a European Union funded FP7-NMP-2009-SME Collaborative Project running from 2010-2014 and involving 13 partners from five different countries. The main objective of this project is to develop new biomass-based composites for use in the automotive industry by adapting existing base biopolymers and developing new biopolymers and new compounding procedures and materials.
The present researcher is involved in the design, development, production, purification and characterisation of a number of novel protein based polymers based on silk and elastin, with the aim of developing and optimising approaches for the efficient industrial scale production and purification of these. This work involves the optimisation of the laboratory scale production (1-3 litre) of these novel polymers in E. coli as well as the scale up and optimisation of the pilot (75 litre, Univ. Minho, Portugal) and industrial scale (500 litre, IBET, Oeiras, Portugal) production and purification processes. Both batch and fed-batch approaches are being investigated.
To date, we have increased production levels to greater than 10 g/L, representing an almost 150-fold increase as compared to production levels previously reported. We have also successfully developed and implemented both pilot and industrial scale fed-batch production and purification processes for these polymers. This work has allowed for a better fundamental understanding of biopolymer production and purification approaches and has enabled the production of large quantities of these polymers which are now being subjected to further characterisation and application studies. This has also led to a number of research articles which are just now beginning to be published.

EcoAgriFood: Innovative Green Products and Processes to Promote Agri-Food Bioeconomy. NORTE-01-0145-FEDER-000009. Norte2020, 2016-2019.

FunBioPlas: Novel synthetic biocomposites for biomedical devices. ERA-IB-2-6/004/2014. ERA-NET, 011016-300919, 2016-2019.

 EngXyl: Unravelling the determinants of xylanase function and stability and enhancing the biotechnological value of a highly active Antarctic xylanase. EXPL/BBB-BIO/1772/2013, Projetos Exploratórios de IC&DT 2013, Fundação para a Ciência e Tecnologia (FCT), Portugal, 070414 – 060415, Principal Investigator.

  NMR Net - National Facility for Nuclear Magnetic Resonance: from Molecular Structure and Dynamics to Protein Function, Cell Physiology, and Metabolomics. RECI/BBB-BQB/0230/2012, Projetos de ICDT 2012, Fundação para a Ciência e Tecnologia (FCT), Portugal, 010713 – 300616, Team Member.

  Psychrophiles and psychrophilic enzymes for the study of compatible solutes and their mechanism of action: the role of protein dynamics. ALTF 950-2006, European Molecular Biology Organisation (EMBO), Heidelberg, Germany, 130507-120509. Principal Investigator.

  Cold-adapted cross-linking enzymes in the food and feed industries (PINDENZ). First Europe Objectif 3, Exercice 2003, EPH3310300R0202, Direction Generale des Technologies de la Recherche et de l’Energie (DGTRE), Belgium, 130505-120507. Principal Investigator.

  Development of a xylanase for use in the food industry (PINDXYL). First Europe Objectif 3, Exercice 2002(2), R0202/215266, Direction Generale des Technologies de la Recherche et de l’Energie (DGTRE), Belgium, 120503-110505. Principal Investigator.

Collins T. and Gerday C. (2017). Enzyme catalysis in psychrophiles. In Margesin R., (Eds), Psychrophiles: From Biodiversity to Biotechnology, Springer International Publishing, Heidelberg, Germany. Ch. 10, pp. 209-235, Edition 2, ISBN 978-3-319-57056-3, DOI 10.1007/978-3-319-57057-0_10.

Barroca M., Santos G., Gerday C. and Collins T. (2017). Biotechnological aspects of cold-active enzymes. In Margesin R., (Eds), Psychrophiles: From Biodiversity to Biotechnology, Springer International Publishing, Heidelberg, Germany. Ch. 19, pp.461-475, Edition 2, ISBN 978-3-319-57056-3, DOI 10.1007/978-3-319-57057-0_19.

Silva D., Santos G., Barroca M. and Collins T. (2017). Inverse PCR for Point Mutation Introduction. In Domingues L. (Ed.) PCR: Methods and Protocols, Series Methods in Molecular Biology. Springer Science + Business Media LLC, New York. Series Vol. 1620. Ch. 5, pp. 87-100, ISBN 978-1-4939-7059-9, DOI 10.1007/978-1-4939-7060-5_5, Series ISSN 1064-3745.

Barroca M., Santos G., Johansson B., Gillotin F., Feller G. and Collins T. (2017). Deciphering the factors defining the pH-dependence of a commercial glycoside hydrolase family 8 enzyme. Enzyme and Microbial Technology, Jan 2017. 96: 163–169. Epub 17/10/16. ISSN 0141-0229. DOI: 10.1016/j.enzmictec.2016.10.011.

Pereira A.M., Machado R., Costa A., Ribeiro A., Bernardo T.C., Collins T., Gomes A.C., Leonor I., Kaplan D.L., Reis R.L. and Casal M. (2017). Silk-based biomaterials functionalized with fibronectin type II promotes cell adhesion. Acta Biomaterialia. Jan 1; 47:50-59. Epub 2016 Oct3. DOI: 10.1016/j.actbio.2016.10.002.

Barroca M., Rodrigues P., Sobral R., Costa M.M.R., Chaves S.R., Machado R., Casal M. and Collins T. Antibiotic free selection for the high level biosynthesis of a silk-elastin-like protein. (2016). Scientific Reports, 6, 39329; DOI: 10.1038/srep39329.

Roulling F., Godin A., Cipolla A., Collins T., Miyazaki K. and Feller G. (2016). Activity–stability relationships revisited in blue oxidases catalyzing electron transfer at extreme temperatures. Extremophiles, 20(5):621-9. DOI 10.1007/s00792-016-0851-9.

Machado R., Costa A., Sencadas V., Pereira A.M., Collins T., Rodríguez-Cabello J.C., Lanceros-Méndez S. and Casal M. (2015). Exploring the Properties of Genetically Engineered Silk-Elastin-Like Protein Films. Macromolecular Bioscience, 15, 1698–1709. DOI: 10.1002/mabi.201500132.

Costa A., Machado M., Ribeiro A., Collins T., Viruthachalam T., Neves-Petersen M.T., Rodríguez-Cabello J.C., Gomes A.C. and Casal M. (2015). Development of Elastin-Like Recombinamer Films with Antimicrobial Activity. Biomacromolecules, 16, 625?635. Epub 2015/01/12. DOI: 10.1021/bm5016706

Carneiro C., Correia A., Collins T., Vilanova M., Pais C., Gomes A.C., Real Oliveira M.E.C.D. and Sampaio P. (2015). DODAB:monoolein liposomes containing Candida albicans cell wall surface proteins: a novel adjuvant and delivery system. European Journal of Pharmaceutics and Biopharmaceutics, 89(2015),190–200. DOI: 10.1016/j.ejpb.2014.11.028.

Collins T., Barroca M., Branca F., Padrão J., Machado R. and Casal M. (2014). High Level Biosynthesis of a Silk-Elastin-like Protein in E. coli. Biomacromolecules, 15(7):2701-8. Epub 2014/06/03. DOI: org/10.1021/bm5005564.

Collins T., Barroca M., Branca F., Azevedo-Silva J., da Costa A., Machado R. and Casal M. (2013). Optimising a cell factory system for the bioproduction of silk-elastin-like polymers. Proceedings of the 1st International Conference on Natural Fibres (ICNF2013): Sustainable Materials for Advanced Applications, pp. 127. Fangueiro R. (Ed.), Guimarães, Portugal. ISBN: 978-989-20-3872-8.

Machado R., da Costa A., Sencadas V., Azevedo-Silva J., Collins T., Arias F.J., Lanceros-Mendez S., Rodríguez-Cabello J.C. and Casal M. (2013). Synthesis and processing of recombinant silk and elastin copolymers. Proceedings of the 1st International Conference on Natural Fibres (ICNF2013): Sustainable Materials for Advanced Applications, pp. 153. Fangueiro R. (Ed.), Guimarães, Portugal. ISBN: 978-989-20-3872-8.

Campos A.R, Oliveira B., Machado R., Collins T., Azevedo-Silva J., da Costa A., Immonen K., Wikström L. and Casal M. (2013). Novel nanocellulose reinforced recombinant protein based polymer composites. Proceedings of the 1^st International Conference on Natural Fibres (ICNF2013): Sustainable Materials for Advanced Applications, pp 333. Fangueiro R. (Ed.), Guimarães, Portugal. ISBN: 978-989-20-3872-8.

Collins T., Azevedo-Silva J., da Costa A., Branca F., Machado R. and Casal M. (2013). Batch production of a silk-elastin-like protein in E. coli BL21(DE3): key stress factors and parameters for optimisation. Microb. Cell Fact., Feb 27; 12:21. DOI: 10.1186/1475-2859-12-21.

Machado M., Correia C., Azevedo-Silva J, Collins T, Arias J, Rodríguez-Cabello J.C. and Casal M. (2013). High level expression and facile purification of recombinant silk-elastin-like polymers in auto induction shake flask cultures. AMB Express, Feb 5; 3(1):11. DOI: 10.1186/2191-0855-3-11.

Pereira A.M., Bernardo T.C., Costa A., Collins T., Machado R., Leonor I.B., Casal M. and Reis R.L. (2013). Development of new chimeric proteins for tissue engineering. J. Tissue Eng. Regen. Med., 7: 1, 46.

Collins T., Feller G., Gerday C. and Meuwis M.A. (2012). Family 8 enzymes with xylanolytic activity. US8309336B2.

Dutron A., Georis J., Genot B., Dauvrin T., Collins T., Hoyoux A. and Feller G. (2011). Use of family 8 enzymes with xylanolytic activity in baking. EP1549147B1(2011), US8192772(2012), CN1681392B(2010), WO2004/023879, PCT/BE03/00152; Granted and published in Europe (all countries in Europe), United States, Mexico, Japan, Eurasia, China, Brazil, Australia.

Collins T., Matzapetakis M. and Santos H. (2010). Backbone and side chain ¹H, ¹⁵N and ¹³C assignments for a thiol-disulphide oxidoreductase from the Antarctic bacterium Pseudoalteromonas haloplanktis TAC125, Biomol. NMR Assign, Vol. 4, No. 2, 151-154.

Marx J.C., D'Amico S., Collins T. and Feller G. (2009). Cold-Adapted Enzymes: Fundamentals and Biotechnological Aspects. In Meshram, S. U. and Shinde, G. B. (eds.), Applied Biotechnology. I.K. International Publishing House Pvt. Limited, pp. 183-188. ISBN: 9380026560, 9789380026565.

Collins T., Roulling F., Florence P., Marx J.C., Feller G., Gerday C. and D’Amico S. (2008). Fundamentals of cold-adapted enzymes. In Margesin R., Schinner F., Marx J.C. and Gerday C. (Eds.), Psychrophiles: From Biodiversity to Biotechnology, Springer-Verlag, Heidelberg, Germany. Ch. 13, pp. 211-227.

Collins T., D'Amico S., Marx J., Feller G. and Gerday C. (2007). Cold-Adapted Enzymes. In Gerday, C. and Glansdorff, N. (eds.), Physiology and biochemistry of extremophiles, ASM Press, Washington, D.C., pp. 165-179.

Marx J.C., Collins T., D'Amico S., Feller G. and Gerday C. (2007). Cold-Adapted Enzymes from Marine Antarctic Microorganisms. Mar. Biotechnol. (NY), Vol. 9, 293–304.

Collins T., Hoyoux A., Van Petegem F., De Vos D., Van Beeumen J., Stals I., Nerinckx W., Claeyssens M., Georis J., Dauvrin T., Arnaut F., Gerday C. and Feller G. (2007.) A novel family 8 psychrophilic xylanase: fundamentals and application. In JAMSTEC, E.R.C. (Ed.), Proceedings of the International Symposium on Extremophiles and their Applications 2005. Japan Science and Technology Information Aggregator, Electronic (J-STAGE), Tokyo, Japan, pp. 73-79. DOI: 10.11490/isea.2005.0.73.0

Gerday C., D’Amico S., Collins T., Marx J.C., Feller G. and Sonan G. (2007). Adaptation strategies and uses of cold adapted enzymes in biotechnological processes. In JAMSTEC, E.R.C. (Ed.), Proceedings of the International Symposium on Extremophiles and their Applications 2005. Japan Science and Technology Information Aggregator, Electronic (J-STAGE), Tokyo, Japan, pp. 66-72. DOI: 10.11490/isea.2005.0.66.0

Collins T., D'Amico S., Georlette D., Marx J.C., Huston A.L. and Feller G. (2006). A nondetergent sulfobetaine prevents protein aggregation in microcalorimetric studies. Anal. Biochem. 352, 299-301.

De Vos D., Collins T., Nerinckx W., Savvides S.N., Claeyssens M., Gerday C., Feller G. and Van Beeumen J. (2006). Oligosaccharide binding in family 8 glycosidases: crystal structures of active-site mutants of the beta-1,4-xylanase pXyl from Pseudoaltermonas haloplanktis TAH3a in complex with substrate and product. Biochemistry, 45, 4797-4807.

Note: De Vos D. and Collins T. contributed equally to this work i.e. joint first authors.

D'Amico S., Collins T., Marx J.C., Feller G. and Gerday C. (2006). Psychrophilic microorganisms: challenges for life. EMBO Rep., 7, 385-389.

Note: All authors contributed equally to this work

Collins T., Hoyoux A., Dutron A., Georis J., Genot B., Dauvrin T., Arnaut F., Gerday C. and Feller G. (2006). Use of glycoside hydrolase family 8 xylanases in baking. JCS, 43, 79-84.

Collins T., De Vos D., Hoyoux A., Savvides S.N., Gerday C., Van Beeumen J. and Feller G. (2005). Study of the active site residues of a glycoside hydrolase family 8 xylanase. J. Mol. Biol., 354, 425-435.

Georis J., Dauvrin T., Hoyoux A., Collins T. and Feller G. (2005). Novel xylanases and their use. World Intellectual Property Organization, PCT, WO 2005/087916 A2.

Collins T., Gerday C. and Feller G. (2005). Xylanases, xylanase families and extremophilic xylanases. FEMS Microbiol. Rev., 29, 3-23.

D’Amico S., Georlette D., Collins T., Feller G. and Gerday C. (2005). Microcalorimetry as applied to psychrophilic enzymes. In Ladbury, J.E. (ed.), Biocalorimetry 2: Application of Calorimetry in the Biological Sciences. Wiley J. and Sons Ltd., The Atrium, Chichester, U.K., pp. 231-240. ISBN: 0470011114, 9780470011119, doi: 10.1002/0470011122.ch13

Hoyoux A., Blaise V., Collins T., D'Amico S., Gratia E., Huston A.L., Marx J.C., Sonan G., Zeng, Y., Feller G. and Gerday C. (2004). Extreme catalysts from low-temperature environments. J. Biosci. Bioeng., 98, 317-330.

Marx J.C., Blaise V., Collins T., D'Amico S., Delille D., Gratia E., Hoyoux A., Huston A.L., Sonan G., Feller G. and Gerday C. (2004). A perspective on cold enzymes: current knowledge and frequently asked questions. Cell. Mol. Biol. (Noisy-le-grand), 50, 643-655. doi: 10.1170/T554

Georlette D., Blaise V., Collins T., D'Amico S., Gratia E., Hoyoux A., Marx J.C., Sonan G., Feller G. and Gerday C. (2004). Some like it cold: biocatalysis at low temperatures. FEMS Microbiol. Rev., 28, 25-42.

Collins T., Meuwis M.A., Gerday C. and Feller G. (2003). Activity, stability and flexibility in glycosidases adapted to extreme thermal environments. J. Mol. Biol., 328, 419-428.

Van Petegem F., Collins T., Meuwis M.A., Gerday C., Feller G. and Van Beeumen J. (2003). The structure of a cold-adapted family 8 xylanase at 1.3 A resolution. Structural adaptations to cold and investigation of the active site. J. Biol. Chem., 278, 7531-7539.

Note: Van Petegem, F. and Collins T. contributed equally to this work i.e. joint first authors.

Van Petegem F., Collins T., Meuwis M.A., Gerday C., Feller G. and Van Beeumen J. (2002). Crystallization and preliminary X-ray analysis of a xylanase from the psychrophile Pseudoalteromonas haloplanktis. Acta Crystallogr. D Biol. Crystallogr., 58, 1494-1496.

Collins T., Meuwis M.A., Stals I., Claeyssens M., Feller G. and Gerday C. (2002). A novel family 8 xylanase: functional and physico-chemical characterization. J. Biol. Chem., 277, 35133-35139.

Collins T., Claverie P., D'Amico S., Georlette D., Gratia E., Hoyoux A., Meuwis M.A., Poncin J., Sonan G., Feller G. and Gerday C. (2002). Life in the cold: psychrophilic enzymes. In Pandalai, S.G. (ed.), Recent Research Developments in Proteins. Transworld Research Network, Trivandrum, Vol. 1, pp. 13-26. ISBN: 81-7895-063-4.

D'Amico S., Claverie P., Collins T., Georlette D., Gratia E., Hoyoux A., Meuwis M.A., Feller G. and Gerday C. (2002). Molecular basis of cold adaptation. Philos. Trans. R. Soc. Lond. B Biol. Sci., 357, 917-925.

Georlette D., Bentahir M., Claverie P., Collins T., D’Amico S., Delille D., Feller G., Gratia E., Hoyoux A., Lonhienne T., Meuwis M.A., Zecchinon L. and Gerday C. (2002). Cold-adapted enzymes. In Bulte J. and Decuyper M. (eds.), Physics and Chemistry Basis of Biotechnology. Vol. 7 of Series Focus on Biotechnology, Kluwer Academic Publishers B.V., pp. 177-196. doi: 10.1007/0-306-46891-3_7.

Zecchinon L., Claverie P., Collins T., D'Amico S., Delille D., Feller G., Georlette D., Gratia E., Hoyoux A., Meuwis M.A., Sonan G. and Gerday C. (2001). Did psychrophilic enzymes really win the challenge? Extremophiles, 5, 313-321.

D'Amico S., Claverie P., Collins T., Feller G., Georlette D., Gratia E., Hoyoux A., Meuwis M.A., Zecchinon L. and Gerday C. (2001). Cold-adapted enzymes: an unachieved symphony. In Storey K.B. and Storey J.M. (eds.), Cell and Molecular Responses To Stress, Vol. 2, Protein Adaptations and Signal Transduction. Elsevier, Amsterdam., Vol. 3, pp. 31-42.

Gerday C., Aittaleb M., Bentahier M., Chessa J.P., Claverie P., Collins T., D'Amico S., Dumont J., Garsoux G., Georlette D., Hoyoux A., Lonhienne T., Meuwis M.A. and Feller G. (2000). Cold-adapted enzymes: from fundamentals to biotechnology. Trends Biotechnol., 18, 103-107.

I have supervised post-doc, PhD, BSc. and MSc. student projects in both Belgium (University of Liege) and Portugal (University of Minho):

Supervision: PhD Ongoing

- Mário Jorge Faria Barroca, PhD in Applied and Environmental Microbiology, University of Minho, Braga, Portugal. Project title: Enhancement of the biotechnological value of a commercial cold-adapted xylanase. Supervisor.

- Augusto Alexandre Costa Barbosa, PhD in Molecular and Environmental Biology, University of Minho, Braga, Portugal. Project title: New generation antigen delivery system for Candida albicans recombinant proteins as a vaccination strategy against systemic fungal infections. Co-supervisor.

Supervision: Research Grantees Completed

- Mário Jorge Faria Barroca, University of Minho, Braga, Portugal. 2014-2015. Unravelling the determinants of xylanase function and stability and enhancing the biotechnological value of a highly active Antarctic xylanase. Research Grantee, FCT Exploratory Project EXPL/BBB-BIO/1772/2013.

Supervision: MSc. Projects Completed

- Augusto Alexandre Costa Barbosa, MSc. in Molecular Genetics, University of Minho, Braga, Portugal. 2017. Development of a novel recombinant subunit antigen as a vaccination target against candidiasis: Cht3 from Candida albicans.

- Gustavo de Almeida Santos, MSc. in Molecular Genetics, University of Minho, Braga, Portugal. 2015. Engineering a commercial cold-adapted xylanase for improved low pH stability.

- Diogo Ribeiro Silva, MSc. in Molecular Genetics, University of Minho, Braga, Portugal. 2015. Understanding adaptation to pH in an industrially relevant glycoside hydrolase family 8 Xylanase

- Paulo Rodrigues, MSc. in Applied Biochemistry, University of Minho, Braga, Portugal. 2014. Genetic approaches for optimising the fed-batch production of a silk elastin-like polymer in E. coli.

- Mário Barroca, MSc. in Biophysics and Bionanosystems., University of Minho, Braga, Portugal. 2013. Optimisation of the fed-batch production of a silk-elastin-like protein: overcoming acetate accumulation and plasmid instability.

- Catherine Oliveira Ferreira, MSc. in Molecular Genetics, University of Minho, Braga, Portugal. 2012. Thioldisulphide oxidoreductases: production, purification and structural analysis of a cold adapted DsbA.

- Frederic Roulling, MSc., University of Liege, Liege, Belgium. 2007. Study of a blue oxidase from the white continent.

- Virginia Brancato, MSc., University of Liege, Liege, Belgium. 2006. pH stability, activity and random mutagenesis of a psychrophilic xylanase.

- Florian Gillotin, MSc., University of Liege, Liege, Belgium, 2005. Study of the activity and stability of a psychrophilic xylanase as a function of pH.

Supervision: BSc. Projects Completed

- Ana Sofia de Oliveira e Silva, BSc. in Applied Biology. University of Minho, Braga, Portugal. 2013. Production, purification and development of an activity assay for a cold adapted DsbA.

- João M. Cunha Ribeiro, BSc. in Applied Biology, University of Minho, Braga, Portugal. 2013. Optimisation of the production of a silk-elastin-like polymer for use in the automobile industry.

- Sophie Leclercq, BSc., University of Liege, Liege, Belgium.2003. Study of a family 8 xylanase: stability and study of the catalytic residues.

- David Gustin, BSc. University of Liege, Liege, Belgium. 2003. Study of a psychrophilic xylanase: thermostabilisation and study of the active site.