title: A New Member of Family 11 Polysaccharide Lyase, Rhamnogalacturonan Lyase (CtRGLf) from Clostridium thermocellum.
abstract: A thermostable, alkaline rhamnogalacturonan lyase (RG lyase) CtRGLf, of family 11 polysaccharide lyase from Clostridium thermocellum was cloned, expressed, purified and biochemically characterised. Both, the full-length CtRGLf (80 kDa) protein and its truncated derivative CtRGL (63.9 kDa) were expressed as soluble proteins and displayed maximum activity against rhamnogalacturonan I (RG I). CtRGLf showed maximum activity at 70 °C, while CtRGL at 60 °C. Both enzymes showed maximum activity at pH 8.5. CtRGLf and CtRGL do not show higher activity on substrates with high β-D-galactopyranose (D-Galp) substitution, this catalytic property deviates from that of some earlier characterised RG lyases which prefer substrates with high D-Galp substitution. The enzyme activity of CtRGLf and CtRGL was enhanced by 1.5 and 1.3 fold, respectively, in the presence of 3 mM of Ca(2+) ions. The TLC analysis of the degraded products of RG I, released by the action of CtRGLf and CtRGL revealed the production of RG oligosaccharides as major products confirming their endolytic activity.
authors.affiliation: Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati, 781039, Assam, India.
authors.forename: Arun
authors.initials: A
authors.lastname: Dhillon
authors.affiliation: CIISA-Faculdade de Medicina Veterinária, Universidade de Lisboa, Av. da Universidade de Técnica, 1300-477, Lisbon, Portugal.
authors.forename: Vania O
authors.initials: VO
authors.lastname: Fernandes
authors.affiliation: CIISA-Faculdade de Medicina Veterinária, Universidade de Lisboa, Av. da Universidade de Técnica, 1300-477, Lisbon, Portugal.
authors.forename: Fernando M V
authors.initials: FM
authors.lastname: Dias
authors.affiliation: CIISA-Faculdade de Medicina Veterinária, Universidade de Lisboa, Av. da Universidade de Técnica, 1300-477, Lisbon, Portugal.
authors.forename: José A M
authors.initials: JA
authors.lastname: Prates
authors.affiliation: CIISA-Faculdade de Medicina Veterinária, Universidade de Lisboa, Av. da Universidade de Técnica, 1300-477, Lisbon, Portugal.
authors.forename: Luis M A
authors.initials: LM
authors.lastname: Ferreira
authors.affiliation: CIISA-Faculdade de Medicina Veterinária, Universidade de Lisboa, Av. da Universidade de Técnica, 1300-477, Lisbon, Portugal.
authors.forename: Carlos M G A
authors.initials: CM
authors.lastname: Fontes
authors.affiliation: CIISA-Faculdade de Medicina Veterinária, Universidade de Lisboa, Av. da Universidade de Técnica, 1300-477, Lisbon, Portugal.
authors.forename: M S J
authors.initials: MS
authors.lastname: Centeno
authors.affiliation: Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati, 781039, Assam, India. arungoyl@iitg.ernet.in.
authors.forename: Arun
authors.initials: A
authors.lastname: Goyal
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002384:Catalysis; D003001:Cloning, Molecular; D048013:Acetivibrio thermocellus / Q000201:enzymology* / Q000235:genetics; D010368:Pectins / Q000378:metabolism; D011133:Polysaccharide-Lyases / Q000235:genetics* / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; C042491:rhamnogalacturonan I; D010368:Pectins; D011133:Polysaccharide-Lyases; D002118:Calcium
keywords: Clostridium thermocellum; Pectic oligosaccharides; Pectin; Polysaccharide lyase; Rhamnogalacturonan; Rhamnogalacturonan Lyase; Rhamnogalacturonan oligosaccharides