TAILIEUCHUNG - Báo cáo khoa học: Characterization of a recombinantly expressed proteinase K-like enzyme from a psychrotrophic Serratia sp.

The gene encoding a peptidase that belongs to the proteinase K family of serine peptidases has been identified from a psychrotrophic Serratiasp., and cloned and expressed inEscherichia coli. The gene has 1890 base pairs and encodes a precursor protein of 629 amino acids with a theoretical molecular mass of kDa. | ềFEBS Journal Characterization of a recombinantly expressed proteinase K-like enzyme from a psychrotrophic Serratia sp. Atle Noralf Larsen1 Elin Moe1 2 Ronny Helland2 Dag Rune Gjellesvik3 and Nils Peder Willassen1 2 1 Department of Molecular Biotechnology Institute of MedicalBiology Faculty of Medicine University of Tromso Norway 2 The Norwegian StructuralBiology Centre University of Tromso Norway 3 Biotec Pharmacon ASA Tromso Norway Keywords Bioprospecting proteinase K like psychrotrophic Serratia sp stability Correspondence N. P. Willassen Department of Molecular Biotechnology Institute of MedicalBiology University of Tromso N-9037 Tromso Norway Tel 47 77 64 46 51 Fax 47 77 64 53 50 E-mail nilspw@ Received 8 September 2005 revised 26 October 2005 accepted 31 October 2005 doi The gene encoding a peptidase that belongs to the proteinase K family of serine peptidases has been identified from a psychrotrophic Serratia sp. and cloned and expressed in Escherichia coli. The gene has 1890 base pairs and encodes a precursor protein of 629 amino acids with a theoretical molecular mass of kDa. Sequence analysis suggests that the peptidase consists of a prepro region a catalytic domain and two C-terminal domains. The enzyme is recombinantly expressed as an active 56 kDa peptidase and includes both C-terminal domains. Purified enzyme is converted to the 34 kDa form by autolytic cleavage when incubated at 50 C for 30 min but retains full activity. In the present work the Serratia peptidase SPRK is compared with the family representative proteinase K PRK from Tritirachium album Limber. Both enzymes show a relatively high thermal stability and a broad pH stability profile. SPRK possess superior stability towards SDS at 50 C compared to PRK. On the other hand SPRK is considerably more labile to removal of calcium ions. The activity profiles against temperature and pH differ for the two enzymes. SPRK shows both a broader pH .

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