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Molecular Characterization of a Fungicidal Endoglucanase from the Cyanobacterium Calothrix elenkinii

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Abstract

A gene responsible for fungicidal activity was identified in the cyanobacterial strain Calothrix elenkinii RPC1, which had shown promise as a biocontrol agent. Functional screening of the genomic library revealed fungicidal (against Pythium aphanidermatum) and endoglucanase activities in two clones. Sequencing revealed an open reading frame of 1,044 bp, encoding 348 amino acid residues with a predicted molecular weight of 38 kDa. Analysis of the deduced amino acid sequence of the putative gene (cael1) showed 99% similarity with the β-1,4-endoglucanase from Anabaena laxa RPAN8 and 97% with the glucanase belonging to the peptidase M20 family of Anabaena variabilis and Nostoc sp. PCC7120, respectively. The putative promoters, ribosomal binding sites and a signal peptide of 22 amino acid residues were identified, revealing the secretory nature of the protein. The phylogenetic tree indicated a close relationship of the gene with Bacillus sp. This study is the first to report on the characterization of an endoglucanase in Calothrix sp.

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Acknowledgments

The authors are thankful to the Post Graduate School and Director, Indian Agricultural Research Institute, New Delhi, India, for providing a fellowship toward the Ph.D. program of the senior author. The study was also partly funded by the AMAAS Network Project on Microorganisms (Theme: Microbial Genomics) granted by the Indian Council of Agricultural Research, New Delhi. We are extremely grateful to Dr. N. K. Singh, National Research Centre for Plant Biotechnology, IARI, New Delhi, for providing the facilities for nebulization of genomic DNA. We are thankful to the Division of Microbiology, IARI, New Delhi, for providing necessary facilities for undertaking this study.

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  1. Vishal Gupta

    Present address: Centre for Cellular and Molecular Biology (CCMB), Council of Scientific and Industrial Research (CSIR), Hyderabad, 500007, Andhra Pradesh, India

Authors and Affiliations

  1. Division of Microbiology, Indian Agricultural Research Institute, New Delhi, 110012, India

    Chitra Natarajan, Vishal Gupta & Radha Prasanna

  2. National Research Centre for Plant Biotechnology, IARI, New Delhi, 110012, India

    Kanika Kumar

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  1. Chitra Natarajan
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Correspondence to Radha Prasanna.

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Supplementary Fig. 1

Identification of promoter gene sequences in cael from RPC1 by BPROM software

Supplementary Fig. 2

Identification of signal peptide and cleavage site by hidden Markov model (HMM). (S2A) Eng from RPC1; (S2B) glucanase precursor protein from Anabaena laxa (ADE22238); (S2C) Anabaena variabilis/Nostoc glucanases (CAA66983/BAB75332). Cleavage site assigned by a probability score together with scores for the n-region, h-region, and c-region of the signal peptide

Supplementary material 3 (DOC 48 kb)

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Natarajan, C., Gupta, V., Kumar, K. et al. Molecular Characterization of a Fungicidal Endoglucanase from the Cyanobacterium Calothrix elenkinii . Biochem Genet 51, 766–779 (2013). https://doi.org/10.1007/s10528-013-9605-x

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