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Psychrophilic trypsin-type protease from Serratia proteamaculans

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Abstract

A preparative method for purification of a novel protease from the psychrotolerant Gram-negative microorganism Serratia proteamaculans (PSP) was developed using affinity chromatography on BPTI-Sepharose. It yielded electrophoretically homogeneous PSP preparation of 60 kD. The PSP properties (temperature and pH stability, high catalytic efficiency) indicate that this enzyme can be defined as a psychrophilic protease. Inhibitory analysis together with substrate specificity indicates that the studied PSP exhibits properties of serine trypsin-like and Zn-dependent protease.

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Abbreviations

ATEE:

Nα-acetyl-L-tyrosine ethyl ester

BAPNA:

Nα-benzoyl-DL-arginine p-nitroanilide

BPTI:

bovine pancreatic trypsin inhibitor

DFP:

diisopropylfluorophosphate

DMSO:

dimethylsulfoxide

DTDP:

4,4′-dithiodipyridine

PMSF:

phenylmethylsulfonyl fluoride

PSP:

Serratia proteamaculans protease

STI:

soybean trypsin inhibitor

TLCK:

Nα-p-tosyl-L-lysyl-chloromethyl-ketone

Z-Lys-S-Bzl:

Nα-benzyloxycarbonyl-L-lysine thiobenzyl ester

buffer A:

0.1 M Tris-HCl, pH 8.0, 50 mM CaCl2

buffer B:

0.1 M Tris-HCl, pH 8.0

buffer C:

0.1 M Tris-HCl, pH 9.0, 50 mM CaCl2, 1 mM MgCl2

buffer D:

10 mM Hepes-KOH, pH 7.5, 1 mM MgCl2

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Authors and Affiliations

  1. Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, 117997, Moscow, Russia

    A. G. Mikhailova, V. V. Likhareva, R. F. Khairullin, N. L. Lubenets & L. D. Rumsh

  2. Institute of Molecular Genetics, Russian Academy of Sciences, pl. Akademika I. V. Kurchatova 2, 123182, Moscow, Russia

    I. V. Demidyuk & S. V. Kostrov

Authors
  1. A. G. Mikhailova
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  2. V. V. Likhareva
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  3. R. F. Khairullin
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  4. N. L. Lubenets
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  5. L. D. Rumsh
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  6. I. V. Demidyuk
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  7. S. V. Kostrov
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Correspondence to A. G. Mikhailova.

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Original Russian Text © A. G. Mikhailova, V. V. Likhareva, R. F. Khairullin, N. L. Lubenets, L. D. Rumsh, I. V. Demidyuk, S. V. Kostrov, 2006, published in Biokhimiya, 2006, Vol. 71, No. 5, pp. 697–706.

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Mikhailova, A.G., Likhareva, V.V., Khairullin, R.F. et al. Psychrophilic trypsin-type protease from Serratia proteamaculans . Biochemistry (Moscow) 71, 563–570 (2006). https://doi.org/10.1134/S0006297906050166

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  • DOI: https://doi.org/10.1134/S0006297906050166

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