Stable linker peptides for a cellulose-binding domain-lipase fusion protein expressed in Pichia pastoris

Fusion proteins composed of a cellulose-binding domain fromNeocallimastix patriciarum cellulase A and Candida antarcticalipase B were constructed using different linker peptides. Theaim was to create proteolytically stable linkers that were ableto join the functional modules without disrupting their function.Six fusion variants containing linkers of 4–44 residueswere expressed in Pichia pastoris and analysed. Three variantswere found to be stable throughout 7-day cultivations. The cellulose-bindingcapacities of fusion proteins containing short linkers wereslightly lower compared with those containing long linkers.The lipase-specific activities of all variants, in solutionor immobilized on to cellulose, were equal to that of the wild-typelipase.