Novel precipitated fluorescent substrates for the screening of cellulolytic microorganisms.
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.
Ivanen
Dina R
DR
Petersburg Nuclear Physics Institute, Russian Academy of Science, Molecular and Radiation Biophysics Division, 188300, Orlova roscha 1, Gatchina, Leningrad District, Russia.
Rongjina
Natalia L
NL
Shishlyannikov
Sergey M
SM
Litviakova
Galina I
GI
Isaeva-Ivanova
Luidmila S
LS
Shabalin
Konstantin A
KA
Kulminskaya
Anna A
AA
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
D052160:Benzothiazoles; D005456:Fluorescent Dyes; D009844:Oligosaccharides; D002482:Cellulose; D044602:Cellulases; D001617:beta-Glucosidase; D043366:Cellulose 1,4-beta-Cellobiosidase