title: Novel precipitated fluorescent substrates for the screening of cellulolytic microorganisms.
abstract: 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, beta-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-beta-d-glucanase and beta-glucosidase.
authors.affiliation: Petersburg Nuclear Physics Institute, Russian Academy of Science, Molecular and Radiation Biophysics Division, 188300, Orlova roscha 1, Gatchina, Leningrad District, Russia.
authors.forename: Dina R
authors.initials: DR
authors.lastname: Ivanen
authors.forename: Natalia L
authors.initials: NL
authors.lastname: Rongjina
authors.forename: Sergey M
authors.initials: SM
authors.lastname: Shishlyannikov
authors.forename: Galina I
authors.initials: GI
authors.lastname: Litviakova
authors.forename: Luidmila S
authors.initials: LS
authors.lastname: Isaeva-Ivanova
authors.forename: Konstantin A
authors.initials: KA
authors.lastname: Shabalin
authors.forename: Anna A
authors.initials: AA
authors.lastname: Kulminskaya
mesh_terms: D001419:Bacteria / Q000201:enzymology / Q000302:isolation & purification*; D052160:Benzothiazoles / Q000737:chemistry; D044602:Cellulases / Q000378:metabolism; D002482:Cellulose / Q000378:metabolism*; D043366:Cellulose 1,4-beta-Cellobiosidase / Q000378:metabolism; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry*; D009844:Oligosaccharides / Q000737:chemistry; D013379:Substrate Specificity; D015003:Yeasts / Q000201:enzymology / Q000302:isolation & purification*; D001617:beta-Glucosidase / Q000378:metabolism
chemical_list: D052160:Benzothiazoles; D005456:Fluorescent Dyes; D009844:Oligosaccharides; D002482:Cellulose; D044602:Cellulases; D001617:beta-Glucosidase; D043366:Cellulose 1,4-beta-Cellobiosidase