Abstract
A new cysteine protease (SpCP) with a molecular mass of about 50 kDa and optimal functioning at pH 8.0 was isolated from the culture medium of a Serratia proteamaculans 94 psychrotolerant strain using affinity and gel permeation chromatography. The enzyme N terminal amino acid sequence (SPVEEAEGDGIVLDV-) exhibits a reliable similarity to N terminal sequences of gingipains R, cysteine proteases from Porphyromonas gingivalis. Unlike gingipains R, SpCP displays a double substrate specificity and cleaves bonds formed by carboxylic groups of Arg, hydrophobic amino acid residues (Val, Leu, Ala, Tyr, and Phe), Pro, and Gly. SpCP can also hydrolyze native collagen. The enzyme catalysis is effective in a wide range of temperatures. Kinetic studies of Z-Ala-Phe-Arg-pNA hydrolysis catalyzed by the protease at 4 and 37°C showed that a decrease in temperature by more than 30°C causes a 1.3-fold increase in the k cat/K m ratio. Thus, SpCP is an enzyme adapted to low positive temperatures. A protease displaying such properties was found in microorganisms of the Serratia genus for the first time and may serve as a virulent factor for these bacteria.
Abbreviations
- A-64:
-
L-trans-3-carboxyoxirane-2-carbonyl-L-leucylagmatine
- Elman’s reagent:
-
5,5′-dithiobis(2-nitrobenzoic acid)
- GPR1 and GPR2:
-
P. gingivalis gingipains R1 and R2, respectively
- MES:
-
2-(N-morpholino)ethanesulfonic acid
- SpCP:
-
S. proteamaculans cysteine protease
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Original Russian Text © N.V. Mozhina, O.A. Burmistrova, D.V. Pupov, G.N. Rudenskaya, Ya.E. Dunaevsky, I.V. Demiduk, S.V. Kostrov, 2008, published in Bioorganicheskaya Khimiya, 2008, Vol. 34, No. 3, pp. 303–309.
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Mozhina, N.V., Burmistrova, O.A., Pupov, D.V. et al. Isolation and properties of Serratia proteamaculans 94 cysteine protease. Russ J Bioorg Chem 34, 274–279 (2008). https://doi.org/10.1134/S1068162008030035
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DOI: https://doi.org/10.1134/S1068162008030035