Novel precipitated fluorescent substrates for the screening of cellulolytic microorganisms

• Published in: Journal of microbiological methods 2009-03, Vol.76 (3), p.295-300
• Main Authors: Ivanen, Dina R., Rongjina, Natalia L., Shishlyannikov, Sergey M., Litviakova, Galina I., Isaeva-Ivanova, Luidmila S., Shabalin, Konstantin A., Kulminskaya, Anna A.
• Format: Article
• Language: English
• Subjects: 2-(2′-Benzothiazolyl)-phenyl cellooligosaccharides; Bacteria - enzymology; Bacteria - isolation & purification; Benzothiazoles - chemistry; beta-Glucosidase - metabolism; Cellulases; Cellulases - metabolism; Cellulose - metabolism; Cellulose 1,4-beta-Cellobiosidase - metabolism; Fluorescent Dyes - chemical synthesis; Fluorescent Dyes - chemistry; Fluorescent precipitated substrates; Oligosaccharides - chemistry; Polymyxa; Substrate Specificity; Yeasts - enzymology; Yeasts - isolation & purification
• ISSN: 0167-7012; 1872-8359
• DOI: 10.1016/j.mimet.2008.12.008

New substrates, 2-(2′-benzothiazolyl)-phenyl ( BTP) cellooligosaccharides with degree of polymerization (d.p.) 2–4 ( BTPG 2–4) were synthesized for the screening of microbial cellulolytic activity in plate assays. The substrates were very efficient that was shown for several cellulolytic bacteria, including yeast-like isolates from Kamchatka hot springs. Three tested bacterial strains and eighteen of 30 of the yeast isolates showed ability to degrade cellulose with cellobiohydrolase, β-glucosidase and endo-cellulase activities measured with standard substrates. The structures of 2-(2′-benzothiazolyl)-phenyl oligosaccharides were solved by NMR- and mass-spectrometry. The usefulness of the 2-(2′-benzothiazolyl)-phenyl substrates were also shown during purification of the B. polymyxa cellulolytic complex, which consists of at least three types of the enzymes: cellobiohydrolase, endo-β- d-glucanase and β-glucosidase.