Stable linker peptides for a cellulose-binding domain-lipase fusion protein expressed in Pichia pastoris.
Fusion proteins composed of a cellulose-binding domain from Neocallimastix patriciarum cellulase A and Candida antarctica lipase B were constructed using different linker peptides. The aim was to create proteolytically stable linkers that were able to join the functional modules without disrupting their function. Six fusion variants containing linkers of 4-44 residues were expressed in Pichia pastoris and analysed. Three variants were found to be stable throughout 7-day cultivations. The cellulose-binding capacities of fusion proteins containing short linkers were slightly lower compared with those containing long linkers. The lipase-specific activities of all variants, in solution or immobilized on to cellulose, were equal to that of the wild-type lipase.
Gustavsson
M
M
Royal Institute of Technology, Stockholm Center for Physics, Astronomy and Biotechnology, Department of Biotechnology, S-10691 Stockholm, Sweden.
Lehtiö
J
J
Denman
S
S
Teeri
T T
TT
Hult
K
K
Martinelle
M
M
D000595:Amino Acid Sequence; D001665:Binding Sites; D002175:Candida / Q000201:enzymology; D002480:Cellulase / Q000737:chemistry*; D002482:Cellulose / Q000378:metabolism*; D004795:Enzyme Stability; D005656:Fungal Proteins; D015870:Gene Expression; D014644:Genetic Variation; D005822:Genetic Vectors; D006031:Glycosylation; D006868:Hydrolysis; D008049:Lipase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D020088:Neocallimastix / Q000201:enzymology; D010455:Peptides / Q000737:chemistry / Q000235:genetics; D010843:Pichia / Q000737:chemistry / Q000235:genetics*; D010957:Plasmids; D015202:Protein Engineering / Q000379:methods*; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D013997:Time Factors
D005656:Fungal Proteins; D010455:Peptides; D011993:Recombinant Fusion Proteins; D002482:Cellulose; D008049:Lipase; C431349:lipase B, Candida antarctica; D002480:Cellulase