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Applied and Environmental Microbiology, February 2004, p. 1135-1144, Vol. 70, No. 2
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.2.1135-1144.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Distribution and Phylogenetic Analysis of Family 19 Chitinases in Actinobacteria

Tomokazu Kawase,¹ Akihiro Saito,² Toshiya Sato,¹ Ryo Kanai,¹ Takeshi Fujii,³ Naoki Nikaidou,¹ Kiyotaka Miyashita,² and Takeshi Watanabe^1*

Department of Applied Biological Chemistry, Faculty of Agriculture, Niigata University, Niigata 950-2181,¹ National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602,² National Institute for Agro-Environmental Sciences, Tsukuba, Ibaraki 305-8604, Japan³

Received 28 July 2003/ Accepted 12 November 2003

In organisms other than higher plants, family 19 chitinase was first discovered in Streptomyces griseus HUT6037, and later, the general occurrence of this enzyme in Streptomyces species was demonstrated. In the present study, the distribution of family 19 chitinases in the class Actinobacteria and the phylogenetic relationship of Actinobacteria family 19 chitinases with family 19 chitinases of other organisms were investigated. Forty-nine strains were chosen to cover almost all the suborders of the class Actinobacteria, and chitinase production was examined. Of the 49 strains, 22 formed cleared zones on agar plates containing colloidal chitin and thus appeared to produce chitinases. These 22 chitinase-positive strains were subjected to Southern hybridization analysis by using a labeled DNA fragment corresponding to the catalytic domain of ChiC, and the presence of genes similar to chiC of S. griseus HUT6037 in at least 13 strains was suggested by the results. PCR amplification and sequencing of the DNA fragments corresponding to the major part of the catalytic domains of the family 19 chitinase genes confirmed the presence of family 19 chitinase genes in these 13 strains. The strains possessing family 19 chitinase genes belong to 6 of the 10 suborders in the order Actinomycetales, which account for the greatest part of the Actinobacteria. Phylogenetic analysis suggested that there is a close evolutionary relationship between family 19 chitinases found in Actinobacteria and plant class IV chitinases. The general occurrence of family 19 chitinase genes in Streptomycineae and the high sequence similarity among the genes found in Actinobacteria suggest that the family 19 chitinase gene was first acquired by an ancestor of the Streptomycineae and spread among the Actinobacteria through horizontal gene transfer.

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* Corresponding author. Mailing address: Department of Applied Biological Chemistry, Faculty of Agriculture, Niigata University, 8050 Ikarashi-2, Niigata 950-2181, Japan. Phone: (81) 25-262-6647. Fax: (81) 25-262-6854. E-mail: wata{at}agr.niigata-u.ac.jp.

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Applied and Environmental Microbiology, February 2004, p. 1135-1144, Vol. 70, No. 2
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.2.1135-1144.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

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