title: A New Subtilase-Like Protease Deriving from Fusarium equiseti with High Potential for Industrial Applications.
abstract: A gene encoding a novel extracellular subtilisin-like protease was cloned from the ascomycete Fusarium equiseti and expressed in Trichoderma reesei. The F. equiseti protease (Fe protease) showed excellent performance in stain removal and good compatibility with several commercial laundry detergent formulations, suggesting that it has high potential for use in various industrial applications. The recombinant enzyme was purified and characterized. The temperature optimum of the Fe protease was 60 °C and it showed high activity in the pH range of 6-10, with a sharp decline in activity at pH above 10. The amino acid specificity of the Fe protease was studied using casein, cytochrome c, and ubiquitin as substrates. The Fe protease had broad substrate specificity: almost all amino acid residues were accepted at position P1, even though it showed some preference for cleavage at the C-terminal side of asparagine and histidine residues. The S4 subsite of Fe protease favors aspartic acid and threonine. The other well-characterized proteases from filamentous fungi, Proteinase K from Engyodontium album, Thermomycolin from Malbranchea sulfurea, and alkaline subtilisins from Bacillus species prefer hydrophobic amino acids in both the S1 and S4 subsites. Due to its different specificity compared to the members of the S8 family of clan SB of proteases, we consider that the Fe protease is a new protease. It does not belong to any previously defined IUBMB groups of proteases.
authors.affiliation: Roal Oy, Tykkimäentie 15, 05200, Rajamäki, Finland, kari.juntunen@roal.fi.
authors.forename: Kari
authors.initials: K
authors.lastname: Juntunen
authors.forename: Susanna
authors.initials: S
authors.lastname: Mäkinen
authors.forename: Sari
authors.initials: S
authors.lastname: Isoniemi
authors.forename: Leena
authors.initials: L
authors.lastname: Valtakari
authors.forename: Alexander
authors.initials: A
authors.lastname: Pelzer
authors.forename: Janne
authors.initials: J
authors.lastname: Jänis
authors.forename: Marja
authors.initials: M
authors.lastname: Paloheimo
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004396:Coloring Agents; D003902:Detergents / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005670:Fusarium / Q000187:drug effects / Q000201:enzymology*; D006861:Hydrogen Peroxide / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007218:Industrial Microbiology; D007700:Kinetics; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D016877:Oxidants / Q000494:pharmacology; D059748:Proteolysis / Q000187:drug effects; D013379:Substrate Specificity / Q000187:drug effects; D013381:Subtilisins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D013696:Temperature; D014242:Trichoderma / Q000201:enzymology
chemical_list: D004396:Coloring Agents; D003902:Detergents; D016877:Oxidants; D006861:Hydrogen Peroxide; D013381:Subtilisins