scholarly journals Chitinolytic enzymes from bacterium inhabiting human gastrointestinal tract -- critical parameters of protein isolation from anaerobic culture.

2011 ◽  
Vol 58 (2) ◽  
Author(s):  
Jarmila Dušková ◽  
Galina Tishchenko ◽  
Evgenia Ponomareva ◽  
Jiří Šimůnek ◽  
Ingrid Koppová ◽  
...  
Keyword(s):  
Gastrointestinal Tract ◽  
Size Exclusion ◽  
Critical Parameters ◽  
Protein Isolation ◽  
Chromatographic Separations ◽  
Healthy Human ◽  
Chitinolytic Enzymes ◽  
Sds Page ◽  
Clostridium Paraputrificum

The object of this study are chitinolytic enzymes produced by bacterium Clostridium paraputrificum J4 isolated from the gastrointestinal tract of a healthy human. In particular, we focus on the development of purification protocols, determination of properties of the enzymes and their activity profiles. The process of bacteria cultivation and isolation of chitinolytic complex of enzymes showing specific activities of endo-, exo-chitinase and N-acetyl-β-glucosaminidase was optimized. A range of various purification procedures were used such as ultrafiltration, precipitation, chromatographic separations (ion-exchange, size exclusion, chromatofocusing) in altered combinations. The optimal purification protocol comprises two or three steps. Individual samples were analyzed by SDS/PAGE electrophoresis and after renaturation their activity could be detected using zymograms. Mass spectroscopy peptide fragment analysis and MALDI analysis of the purest samples indicate presence of endochitinase B (molecular mass about 85 kDa) and of 60-kDa endo- and exochitinases.

scholarly journals Determination of molecular masses for petroleum distillates by simualted distillation

2012 ◽  
Vol 4 (5) ◽  
pp. 43-55 ◽  
Author(s):  
Lante Carbognani
Keyword(s):  
Stationary Phases ◽  
Size Exclusion ◽  
Chromatographic Separations ◽  
Calibration Standards ◽  
Hydrocarbon Group ◽  
Simulated Distillation ◽  
Size Exclusion Chromatographic ◽  
Petroleum Distillates ◽  
Molecular Masses

Determination of Molecular Mass (MM) for petroleum distillates is explored for selected samples viaHigh Temperature Simulated Distillation (HTSD). MM is determined as a by-product from routineHTSD carried out using open wall capillary columns coated with apolar stationary phases. No samplepre-separation into hydrocarbon group-types is required. Determined MM values were validated with resultsachieved via correlations based on specific gravity and refractive index. Furthermore, determined MM withthe former methodologies were found to match mass spectrometric determinations carried out for the basicpolar components present within the studied samples. Moreover, HTSD analyzed petroleum distillates areproposed as more representative calibration standards for size exclusion chromatographic separations, thusopening the feasibility of extrapolating MM determination for non volatile petroleum residua.

scholarly journals Biosynthesis of mucins (MUC2-6) along the longitudinal axis of the human gastrointestinal tract

1997 ◽  
Vol 273 (2) ◽  
pp. G296-G302 ◽  
Author(s):  
B. J. Van Klinken ◽  
J. Dekker ◽  
H. A. Buller ◽  
C. de Bolos ◽  
A. W. Einerhand
Keyword(s):  
Gastrointestinal Tract ◽  
Large Intestine ◽  
Allelic Variation ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Longitudinal Axis ◽  
Human Gastrointestinal Tract ◽  
Healthy Human ◽  
Sds Page ◽  
Small And Large Intestine

Little is known about the biosynthesis of mucin molecules in humans. Our aim was to examine the mucin biosynthesis (MUC2-6) along the longitudinal axis of the healthy human gastrointestinal tract. Biopsies of human stomach and small and large intestine were metabolically labeled with 35S-labeled amino acids, [35S]sulfate, or[3H]galactose, immunoprecipitated with antibodies against MUC2-6, and analyzed by reducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), MUC5AC [apparent molecular weight (M(r)) 500,000] and MUC6 (apparent M(r) 400,000) were detected in the stomach but not in the small or large intestine, MUC3 (apparent M(r) 550,000) was detected in duodenum and jejunum, MUC2 (apparent M(r)600,000) was detected throughout the small and large intestine, and MUC4 (apparent M(r) > 900,000) was detected predominantly in the large intestine. Interestingly, some individuals displayed double bands of MUC2 and MUC3 precursors, suggesting allelic variation within the respective genes. Between small and large intestine mature secreted MUC2 showed differences in mobility on SDS-PAGE, suggesting differences in glycosylation. Each of the MUC2, MUC3, MUC4, MUC5AC, and MUC6 precursors could be distinguished electrophoretically, and each showed region-specific expression along the gastrointestinal tract.

Sign in / Sign up

Export Citation Format

Share Document