| Title : Oligopeptidase B from Serratia proteamaculans. I. Determination of primary structure, isolation, and purification of wild-type and recombinant enzyme variants - Khairullin_2009_Biochemistry.(Mosc)_74_1164 |
| Author(s) : Khairullin RF , Mikhailova AG , Sebyakina TY , Lubenets NL , Ziganshin RH , Demidyuk IV , Gromova TY , Kostrov SV , Rumsh LD |
| Ref : Biochemistry (Mosc) , 74 :1164 , 2009 |
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Abstract :
A novel trypsin-like protease (PSP) from the psychrotolerant gram-negative microorganism Serratia proteamaculans was purified by ion-exchange chromatography on Q-Sepharose and affinity chromatography on immobilized basic pancreatic trypsin inhibitor (BPTI-Sepharose). PSP formed a tight complex with GroEL chaperonin. A method for dissociating the GroEL-PSP complex was developed. Electrophoretically homogeneous PSP had molecular mass of 78 kDa; the N-terminal amino acid sequence 1-10 was determined, and mass-spectral analysis of PSP tryptic peptides was carried out. The enzyme was found to be the previously unknown oligopeptidase B (OpdB). The S. proteamaculans 94 OpdB gene was sequenced and the producer strain Escherichia coli BL-21(DE3) pOpdB No. 22 was constructed. The yield of expressed His(6)-PSP was 1.5 mg/g biomass. |
| PubMedSearch : Khairullin_2009_Biochemistry.(Mosc)_74_1164 |
| PubMedID: 19916930 |
| Gene_locus related to this paper: 9gamm-b3vi58 |
| Gene_locus | 9gamm-b3vi58 |
Khairullin RF, Mikhailova AG, Sebyakina TY, Lubenets NL, Ziganshin RH, Demidyuk IV, Gromova TY, Kostrov SV, Rumsh LD (2009)
Oligopeptidase B from Serratia proteamaculans. I. Determination of primary structure, isolation, and purification of wild-type and recombinant enzyme variants
Biochemistry (Mosc)
74 :1164
Khairullin RF, Mikhailova AG, Sebyakina TY, Lubenets NL, Ziganshin RH, Demidyuk IV, Gromova TY, Kostrov SV, Rumsh LD (2009)
Biochemistry (Mosc)
74 :1164