title: Heat-labile proteases in molecular biology applications.
abstract: Thermolabile proteases were identified in three Gram-negative psychrotrophic bacteria. The protease from the psychrotrophic strain A9 was purified and its application to common molecular biology techniques was demonstrated. Heat-stable molecular biology enzymes (Taq polymerase and PvuII) were digested by a heat-labile protease, which was then inactivated by a mild heat treatment. The clear benefit of using heat-labile proteases arises in situations where further reactions may be accomplished without an intermediate purification step, thereby saving time and avoiding the possibility of product loss.
authors: Department of Biochemistry and Microbiology, University of Victoria, P.O. Box 3055, V8W 3P6, Victoria, BC, Canada.
authors: A J
authors: AJ
authors: Moran
authors: M
authors: M
authors: Hills
authors: J
authors: J
authors: Gunton
authors: F E
authors: FE
authors: Nano
mesh_terms: D003080:Cold Temperature; D003470:Culture Media; D004269:DNA, Bacterial / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D010450:Endopeptidases / Q000302:isolation & purification* / Q000378:metabolism*; D004795:Enzyme Stability / Q000502:physiology; D006090:Gram-Negative Bacteria / Q000201:enzymology* / Q000254:growth & development; D006358:Hot Temperature; D008967:Molecular Biology / Q000379:methods*; D016133:Polymerase Chain Reaction; D012623:Seawater / Q000382:microbiology; D019914:Taq Polymerase / Q000378:metabolism
chemical_list: D003470:Culture Media; D004269:DNA, Bacterial; D019914:Taq Polymerase; C025630:CAGCTG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific; D010450:Endopeptidases