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Purification and Partial Characterization of a Novel β-1,3-Endoglucanase from Streptomyces rutgersensis

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Abstract

A β-1,3-endoglucanase produced by Streptomyces rutgersensis was purified to a homogeneity by the fractional precipitation with ammonium sulfate, ion exchange chromatography on Q-Sepharose and hydrophobic chromatography on Butyl Sepharose. A typical procedure provided 11.74-fold purification with 12.53 % yield. SDS-PAGE of the purified protein showed one protein band. The exact molecular mass of the enzyme obtained by mass spectrometry was 41.25 kDa; the isoelectric point was between pH 4.2–4.4. The optimal β-glucanase catalytic activity was at pH 7 and 50 °C. An enzyme was only active toward glucose polymers containing β-1,3 linkages and hydrolyzed Saccharomyces cerevisiae cell wall β-glucan in an endo-like way: reaction products were different molecular size β-glucans, which were larger than glucose.

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Abbreviations

AU:

Unit of enzyme activity

ESI:

Electrospray ionization

Km :

Michaelis–Menten constant

MS:

Mass spectrometry

pI:

Isoelectric point

QTOF:

Quadrupole time-of-flight mass spectrometer

RPM:

Revolutions per minute

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

SEC:

Size-exclusion chromatography

TLC:

Thin layer chromatography

Vmax :

Maximum velocity

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Authors and Affiliations

  1. JSC “Biocentras”, Graičiūno St. 10, 02241, Vilnius, Lithuania

    Artur Javmen & Saulius Grigiškis

  2. Department of Immunology, State Scientific Research Institute Centre for Innovative Medicine, Moletu Pl. 29, 08409, Vilnius, Lithuania

    Artur Javmen & Mykolas Mauricas

  3. Department of Chemistry and Bioengineering, Vilnius Gediminas Technical University, Saulėtekio St. 11, 10223, Vilnius, Lithuania

    Mark Rudenkov

Authors
  1. Artur Javmen
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  2. Saulius Grigiškis
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  3. Mark Rudenkov
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  4. Mykolas Mauricas
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Correspondence to Artur Javmen.

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Javmen, A., Grigiškis, S., Rudenkov, M. et al. Purification and Partial Characterization of a Novel β-1,3-Endoglucanase from Streptomyces rutgersensis . Protein J 32, 411–417 (2013). https://doi.org/10.1007/s10930-013-9500-7

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