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Cellulase activity of a haloalkaliphilic anaerobic bacterium, strain Z-7026

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Summary

The cellulolytic activity of an alkaliphilic obligate anaerobic bacterium, Z-7026, which was isolated from the microbial community of soda-lake sediments and belongs to the cluster III of Clostridia with low G+C content, was studied. The bacterium was capable of growing in media with cellulose or cellobiose as the sole energy sources. Its maximal growth rate on cellobiose (0.042–0.046 h−1) was observed at an initial pH value of 8.5–9.0, whereas the maximal rate of cellulase synthesis, assayed by using a novel fluorimetric approach, was found to be 0.1 h−1 at pH 8–8.5. Secreted proteins revealed high affinity for cellulose and were represented by two major forms of molecular masses of 75 and 84 kDa, whereas the general protein composition of the precipitated and cellulose-bound preparations was similar to cellulosome subunits of Clostridium thermocellum. The optimum pH of the partially purified enzyme preparation towards both amorphous and crystalline cellulose was in the range 6–9, with more than 70% and less than 50% of maximal activity being retained at pH 9.2 and 5.0, respectively.

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Abbreviations

B:

Protein band in the electrophoretic separation

BSA:

bovine serum albumin

Cbh:

Cellobiohydrolase

Cel:

Cellulase

CBM:

Cellulose-binding module

CPB:

Citrate phosphate buffer

GH:

Glycosyl hydrolase

MCC:

Microcrystalline cellulose

MUF:

4-methylumbelliferon

MUFC:

4-methylumbelliferyl-β-D-cellobioside

MUFL:

4-methylumbelliferyl-β-D-lactopyranoside

PASC:

Phosphoric acid-swollen cellulose

S:

Protein subunit of the C. thermocellum cellulosome

SDS-PAGE:

Electrophoresis in polyacrylamide gel in the presence of sodium dodecyl sulfate

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Acknowledgements

The authors thank L. G. Vasilchenko, K. N. Karapetyan, and V.V. Khromonygina for qualified assistance and fruitful discussion. Critical reading of the manuscript by professor D. Haltrich is also gratefully acknowledged. This work was supported by grants RFBR 02-04-48286 and 02-04-49033, and the Program “Molecular and Cellular Biology” of Russian Academy of Sciences.

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Authors and Affiliations

  1. A. N. Bach Institute of Biochemistry, Russian Academy of Sciences, 33, Leninsky prospect, 119071, Moscow, Russia

    E. A. Zvereva, T. V. Fedorova & M. L. Rabinovich

  2. Winogradsky Institute of Microbiology, Russian Academy of Sciences, 7/2, Prospect 60-letija Octiabria, 117312, Moscow, Russia

    V. V. Kevbrin & T. N. Zhilina

Authors
  1. E. A. Zvereva
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  2. T. V. Fedorova
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  3. V. V. Kevbrin
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  4. T. N. Zhilina
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  5. M. L. Rabinovich
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Correspondence to M. L. Rabinovich.

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Communicated by K. Horikoshi

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Zvereva, E.A., Fedorova, T.V., Kevbrin, V.V. et al. Cellulase activity of a haloalkaliphilic anaerobic bacterium, strain Z-7026. Extremophiles 10, 53–60 (2006). https://doi.org/10.1007/s00792-005-0472-1

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