title: Cellulase-xylanase synergy in designer cellulosomes for enhanced degradation of a complex cellulosic substrate.
abstract: Designer cellulosomes are precision-engineered multienzyme complexes in which the molecular architecture and enzyme content are exquisitely controlled. This system was used to examine enzyme cooperation for improved synergy among Thermobifida fusca glycoside hydrolases. Two T. fusca cellulases, Cel48A exoglucanase and Cel5A endoglucanase, and two T. fusca xylanases, endoxylanases Xyn10B and Xyn11A, were selected as enzymatic components of a mixed cellulase/xylanase-containing designer cellulosome. The resultant mixed multienzyme complex was fabricated on a single scaffoldin subunit bearing all four enzymes. Conversion of T. fusca enzymes to the cellulosomal mode followed by their subsequent incorporation into a tetravalent cellulosome led to assemblies with enhanced activity (~2.4-fold) on wheat straw as a complex cellulosic substrate. The enhanced synergy was caused by the proximity of the enzymes on the complex compared to the free-enzyme systems. The hydrolytic properties of the tetravalent designer cellulosome were compared with the combined action of two separate divalent cellulase- and xylanase-containing cellulosomes. Significantly, the tetravalent designer cellulosome system exhibited an ~2-fold enhancement in enzymatic activity compared to the activity of the mixture of two distinct divalent scaffoldin-borne enzymes. These results provide additional evidence that close proximity between cellulases and xylanases is key to the observed concerted degradation of the complex cellulosic substrate in which the integrated enzymes complement each other by promoting access to the relevant polysaccharide components of the substrate. The data demonstrate that cooperation among xylanases and cellulases can be augmented by their integration into a single designer cellulosome.
authors.affiliation: Department of Biological Chemistry, The Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Sarah
authors.initials: S
authors.lastname: Moraïs
authors.forename: Yoav
authors.initials: Y
authors.lastname: Barak
authors.forename: Jonathan
authors.initials: J
authors.lastname: Caspi
authors.forename: Yitzhak
authors.initials: Y
authors.lastname: Hadar
authors.forename: Raphael
authors.initials: R
authors.lastname: Lamed
authors.forename: Yuval
authors.initials: Y
authors.lastname: Shoham
authors.forename: David B
authors.initials: DB
authors.lastname: Wilson
authors.forename: Edward A
authors.initials: EA
authors.lastname: Bayer
mesh_terms: D000192:Actinomycetales / Q000201:enzymology* / Q000235:genetics; D001709:Biotechnology / Q000379:methods; D002480:Cellulase / Q000235:genetics* / Q000378:metabolism*; D044603:Cellulosomes / Q000235:genetics* / Q000378:metabolism*; D043364:Endo-1,4-beta Xylanases / Q000235:genetics* / Q000378:metabolism*; D007700:Kinetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014908:Triticum / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D002480:Cellulase; D043364:Endo-1,4-beta Xylanases