| Title : Cloning, sequencing, expression, and characterization of thermostability of oligopeptidase B from Serratia proteamaculans, a novel psychrophilic protease - Mikhailova_2014_Protein.Expr.Purif_93_63 |
| Author(s) : Mikhailova AG , Khairullin RF , Demidyuk IV , Kostrov SV , Grinberg NV , Burova TV , Grinberg VY , Rumsh LD |
| Ref : Protein Expr Purif , 93 :63 , 2014 |
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Abstract :
Protease from Serratia proteamaculans (PSP) is the first known psychrophilic oligopeptidase B. The gene of S. proteamaculans 94 oligopeptidase B was cloned, sequenced and expressed in Escherichia coli. The unfolding of PSP molecule following heat treatment at 37 degC by measuring fluorescence spectra was examined in parallel with the residual activity determination. The effect of PSP thermostabilization by glycerol at 37-50 degree was revealed. Calcium ions and buffer solution of low molarity cause the opposite effect - the acceleration of PSP inactivation at 37 degC. The thermal stability of PSP molecule in the presence of 0-100mM CaCl2 was also investigated by means of high-sensitivity differential scanning calorimetry. The artificial reconstruction of the natural complex PSP-chaperonin from S. roteamaculans was carried out: the stable complex (1:1) of chaperonin E. oli GroEL with active recombinant enzyme PSP was obtained. It was shown that complex formation with chaperonin promotes PSP thermostability at 37 degC. |
| PubMedSearch : Mikhailova_2014_Protein.Expr.Purif_93_63 |
| PubMedID: 24184401 |
| Gene_locus related to this paper: 9gamm-b3vi58 |
| Gene_locus | 9gamm-b3vi58 |
Mikhailova AG, Khairullin RF, Demidyuk IV, Kostrov SV, Grinberg NV, Burova TV, Grinberg VY, Rumsh LD (2014)
Cloning, sequencing, expression, and characterization of thermostability of oligopeptidase B from Serratia proteamaculans, a novel psychrophilic protease
Protein Expr Purif
93 :63
Mikhailova AG, Khairullin RF, Demidyuk IV, Kostrov SV, Grinberg NV, Burova TV, Grinberg VY, Rumsh LD (2014)
Protein Expr Purif
93 :63