Stabilization of Taq DNA polymerase at high temperature by protein folding pathways from a hyperthermophilic archaeon, Pyrococcus furiosus.
Pyrococcus furiosus, a hyperthermophilic archaeon growing optimally at 100 degrees C, encodes three protein chaperones, a small heat shock protein (sHsp), a prefoldin (Pfd), and a chaperonin (Cpn). In this study, we report that the passive chaperones sHsp and Pfd from P. furiosus can boost the protein refolding activity of the ATP-dependent Cpn from the same hyperthermophile. The thermo-stability of Taq polymerase was significantly improved by combinations of P. furiosus chaperones, showing ongoing protein folding activity at elevated temperatures and during thermal cycling. Based on these results, we propose that the protein folding apparatus in the hyperthermophilic archaeon, P. furiosus can be utilized to enhance the durability and cost effectiveness of high temperature biocatalysts.
Laksanalamai
Pongpan
P
Center of Marine Biotechnology, University of Maryland Biotechnology Institute, 701 E. Pratt St., Baltimore, MD 21202, USA. pongpan@comb.umbi.umd.edu
Pavlov
Andrey R
AR
Slesarev
Alexei I
AI
Robb
Frank T
FT
D019843:Archaeal Proteins / Q000737:chemistry*; D018833:Chaperonins; D050888:Heat-Shock Proteins, Small / Q000737:chemistry*; D006358:Hot Temperature; D018832:Molecular Chaperones / Q000737:chemistry*; D017510:Protein Folding; D020027:Pyrococcus furiosus / Q000737:chemistry*; D019914:Taq Polymerase / Q000737:chemistry*
D019843:Archaeal Proteins; D050888:Heat-Shock Proteins, Small; D018832:Molecular Chaperones; C112806:prefoldin; D019914:Taq Polymerase; D018833:Chaperonins