01996nas a2200241   4500008004100000022001400041245019800055210006900253260009500322300001200417490000800429520105500437653001401492653000801506653000901514653002301523653002401546653001001570653001301580100002101593700001101614856012901625       2005                        eng d  a0006-291X00aPurification and properties of a low molecular weight 1,4-beta-D-glucan glucohydrolase having one active site for carboxymethyl cellulose and xylan from an alkalothermophilic Thermomonospora sp0 aPurification and properties of a low molecular weight 14betaDglu  a525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USAbACADEMIC PRESS INC ELSEVIER SCIENCEcAPR  a111-1160 v3293 a<p>A low molecular weight 1,4-beta-D-glucan glucohydrolase from an extracellular culture filtrate of Thermomonospora sp. was purified to homogeneity. The molecular weight of the purified enzyme was 14.2 kDa by MALDI-TOF analysis and is in agreement with SDS-PAGE and gel filtration chromatography. The purified enzyme exhibited both endocarboxymethyl cellulase and endoxylanase activities. A kinetic method was employed to study the active site of the enzyme that hydrolyzes both carboxymethyl cellulose and xylan. The experimental data coincide well with the theoretical values calculated for the case of a single active site. Conformation and microenvironment at the active site was probed with fluorescent chemo-affinity labeling using o-phthalaidehyde as the chemical initiator. Formation of isoindole derivative resulted in complete inactivation of the enzyme to hydrolyze both xylan and CMC as judged by fluorescence studies corroborating a single active site for the hydrolysis of xylan and CMC. (C) 2005 Elsevier Inc. All rights reserved.</p>10aCellulase10aCMC10aOPTA10asingle active site10aThermomonospora sp.10axylan10aXylanase1 aJagtap, Sharmili1 aRao, M  uhttps://library.ncl.res.in/content/purification-and-properties-low-molecular-weight-14-beta-d-glucan-glucohydrolase-having-002180nas a2200205   4500008004100000022001400041245018700055210006900242260007500311300001200386490000700398520133500405653000801740653001401748653002801762653002001790100002101810700001401831856012901845       2009                        eng d  a1053-050900aFluorescence study on interactions of alpha-crystallin with the molten globule state of 1, 4-beta-D-glucan glucanohydrolase from thermomonospora sp induced by guanidine hydrochloride0 aFluorescence study on interactions of alphacrystallin with the m  a233 SPRING ST, NEW YORK, NY 10013 USAbSPRINGER/PLENUM PUBLISHERScNOV  a967-9730 v193 a<p>In this paper, the interaction between alpha- crystallin and molten globule structure of 1,4-beta-D-Glucan Glucohydrolase (TSC) from an alkalothermophilic Thermomonospora sp. was investigated mainly by fluorescence quenching spectra, circular dichroism and three dimensional fluorescence spectra under simulative physiological conditions. Denaturation studies using GdnCl indicated that TSC folds through a partially folded state that resembles molten globule at 1.8 M GdnCl. The chaperone activity of alpha- crystallin was employed to study refolding of TSC. Here we studied the refolding of GdnCl denatured TSC from its molten globule state (TSC-m complex) in the presence and absence of alpha-crystallin to elucidate the molecular mechanism of chaperone-mediated in vitro folding. Our results, based on intrinsic tryptophan fluorescence and ANS binding studies, suggest that alpha-crystallin formed a complex with a putative intermediate molten globule - like intermediate in the refolding pathway of TSC. Reconstitution of the active TSC was observed on cooling the alpha-crystallin aEuro cent TSC -m complex to 4A degrees C. Addition of alpha-crystallin to the molten globule - like intermediate of TSC (TSC-m complex) complex initiated the refolding of TSC with 69 % recovery of the biological activity of the enzyme.</p>10aANS10aCellulase10aGuanidine hydrochloride10aProtein folding1 aJagtap, Sharmili1 aRao, Mala  uhttps://library.ncl.res.in/content/fluorescence-study-interactions-alpha-crystallin-molten-globule-state-1-4-beta-d-glucan-0