ABSTRACT
Alkaline xylanases from alkaliphilic Bacillus strains NCL (87-6-10) and Sam III were compared with the commercial xylanases Pulpzyme HC and Biopulp for their compatibility with detergents and proteases for laundry applications. Among the four xylanases evaluated, the enzyme from the alkaliphilic Bacillus strain NCL (87-6-10) was the most compatible. The enzyme retained its full activity (40 degrees C for 1 h) in the presence of detergents, whereas Pulpzyme HC and Sam III showed only 30% and 50% of their initial activity, respectively. Biopulp, though stable to detergents, had only marginal activity (5%)at pH 10. However, all four enzymes retained significant activity (80%) for 60 min in the presence of the proteases Alcalase and Conidiobolus protease. Supplementation of the enzyme enhanced the cleaning ability of the detergents.
Subject(s)
Bacillus/enzymology , Detergents , Endo-1,4-beta Xylanases/metabolism , Endopeptidases/metabolism , Industrial Microbiology/methods , Alkalies , Hydrogen-Ion Concentration , Paper , WoodABSTRACT
Two alkaline xylanases designated as "A" and "C", respectively, were isolated from the culture filtrates of the alkalophilic Bacillus grown on a wheat bran-yeast extract medium. The two xylanases occurred in the culture filtrate in a ratio of 10:90. These xylanases were purified to homogeneity on a CM-Sephadex matrix followed by further separation of Xylanase "A" on a phenyl sepharose column and preparative electrophoresis. The two xylanases differed considerably in their physico-chemical properties, kinetics and in their mode of action. Xylanase "C" had a molecular weight of 25,000 as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis and was a cationic protein with a pI of 8.9. In contrast xylanase "A" had a molecular weight of 45,000 with a pI of 5.3. The two xylanases showed distinct differences in their hydrolysis pattern. Xylanase "A" produced comparatively larger amounts of small molecular weight oligosaccharides and xylose namely xylotriose (X(3)), xylobiose (X(2)) and xylose even in the initial stages of hydrolysis (2 and 5 h) while xylanase "C" produced negligible amounts of X(2) and no xylose for the same period of incubation. At 24 h only traces of xylose was produced by xylanase "C" while substantial amounts of the monomer was produced by xylanase A in 24 h. Xylanase "A" had a broad pH optimum ranging from pH 6.0-10.0 at 40-60 degrees C while xylanase "C" had an optimum pH of 8.0 at 40-60 degrees C. Xylanases "A" and "C" differed in their K(m) and V(max) values. Xylanase "A" had a K(m) of 1.67 mg/ml and a V(max) of 3.85 x 10(2) micromol/ml/min, whereas xylanase "C" had a K(m) of 10 mg/ml and a V(max) of 1.43 x 10(4) micromol/ml/min.
Subject(s)
Bacillus/enzymology , Xylosidases/isolation & purification , Xylosidases/metabolism , Cellulase/isolation & purification , Chromatography, Ion Exchange , Electrophoresis, Polyacrylamide Gel , Isoenzymes/chemistry , Isoenzymes/isolation & purification , Isoenzymes/metabolism , Kinetics , Molecular Weight , Substrate Specificity , Xylan Endo-1,3-beta-Xylosidase , Xylosidases/chemistryABSTRACT
An alkalophilicBacillus (NCL-87-6-10, NCIM 2128), with a high productivity for extracellular xylanase (EC 3.2.1.8) and free of cellulase, was isolated from soil containing coconut fibre detritus. When grown on a wheat bran/yeast extract medium in submerged culture for 48 h, it produced 100 to 120 IU of enzyme activity per ml. The crude enzyme consists of two fractions of apparent mol sizes of approx 10.4 and 29 kDa in the proportion of 90:10, as determined by native gel exclusion chromatography. Optimum activity of the xylanase was at 60°C and pH 8.0. A two-fold increase in enzyme activity was obtained when reducing agents, thioethanol and dithiothreitol, were included in the assay.
ABSTRACT
A method for the location of proteins/enzymes by polyacrylamide gel electrophoresis using dialyzable low-molecular-weight fluorescent peptide markers is described. The markers prepared by treating the peptic digest of casein with fluorescamine showed several bands on gel electrophoresis which helped in locating the proteins. The located desired protein could subsequently be purified by extraction.
Subject(s)
Aldose-Ketose Isomerases , Carbohydrate Epimerases/isolation & purification , Electrophoresis, Polyacrylamide Gel , Fluorescent Dyes , Serum Albumin, Bovine/isolation & purification , Subtilisins/antagonists & inhibitors , Dialysis , Fluorescamine , Fluorescence , Plant Proteins/isolation & purification , Plants/analysis , Streptomyces/enzymologyABSTRACT
A simple, inexpensive acrylate polymer which has a capacity to absorb 170 ml of water per g has been developed. It can be used to concentrate dilute solutions of macromolecules such as proteins, nucleic acids, and carbohydrates. The polymer absorbs only low-molecular-weight substances such as glucose, sucrose, and inorganic salts. It can replace the various conventional concentration methods. No special device or electricity is needed for the concentration. The inexpensive polymer, molded in the form of rods, can be very conveniently used as "disposable concentration sticks."
