VIEW ARTICLE    DOI: 10.1094/ASBCJ-56-0019

Characterization of Pectinatus cerevisiiphilus and P. frisingensis by Ribotyping (1). Yasuo Motoyama (2), Tomoo Ogata, and Kazuhisa Sakai, Brewing Research & Development Laboratory, Asahi Breweries Ltd., 1-21, Midori, 1-Chome, Moriya-machi, Kitasoma-gun, lbaraki, 302-0106 Japan. (1) Presented at the 63rd ASBC Annual Meeting, Palm Springs, CA, June 1997. (2) Corresponding author. Phone: +81-297-46-1511. Fax: +81-297-46-1514. E-mail: <yasuo.motoyama@asahibeer.co.jp> J. Am. Soc. Brew. Chem. 56(1):19-23, 1998. Accepted December 18, 1997.

A total of 39 Pectinatus strains have been ribotyped in this investigation. Thirteen different ribotypes were observed from 34 strains of P. frisingensis, while three different types were shown with five strains of P. cerevisiiphilus when digested with EcoRI. These patterns consisted of two series of fragments: fragments conserved within a species, and polymorphic variation. P. frisingensis had conserved fragments of approximately 2.5-3, 7-8, and 10-20 kbp, whereas P. cerevisiiphilus had approximately 2-3, 5-6, and 20-30 kbp. These fragment patterns, characterized by sets of conserved fragments within a species, were species specific. Furthermore, the HindIII- and BamHI-digested restriction patterns have been examined to facilitate differentiation at the strain level. Thirty-four strains of P. frisingensis could be divided into 1 of 12 types. On the other hand, a composite ribotype, which combined the ribotypes produced by all restriction enzymes used, produced such high definition that all strains of P. frisingensis fell into 1 of 17 types. Keywords: Beer-spoilage bacteria, Pectinatus, Restriction fragment length polymorphism (RFLP), Ribosomal RNA, Ribotyping.

Un total de 39 cepas de Pectinatus han sido clasificadas en ribotipos en esta investigación. Trece ribotipos diferentes fueron observados de 34 cepas de P. frisingensis mientras tres tipos diferentes fueron mostrados con cinco cepas de P. cerevisiiphilus cuando se digirieron con EcoRI. Estos patrones consistieron de dos series de fragmentos: fragmentos conservados dentro de una especie, y una variación polimórfica. P. frisingensis ha conservado fragmentos de aproximar 2.5-3, 7-8, y 10-20 kbp, en donde P. cerevisiiphilus tuvo aproximar 2-3, 5-6, y 20-30 kbp. Estos fragmentos patrones, caracterizados por juegos de fragmentos conservados dentro de una especie, fueron especie-específicos. Además, los patrones de restricción digeridos HindIII y BamHI han sido examinados para facilitar la diferenciación a nivel de cepa. Cepas (34) de P. frisingensis podían estar dividas en uno de doce tipos. Por otra parte, un ribotipo compuesto el cual combinó los ribotipos producidos por toda las enzimas de restricción usadas, produjeron tan alta definición que todas las cepas de P. frisingensis estaban comprendidas en uno de los 17 tipos.