Glucosidase isolation

All assays (21) contained C 180 / g protein, 1000 units of Cx (CM-cellulose) activity (26), and 100 units of /3-glucosidase where necessary, /3-glucosidase isolated from the cellulase of F. solani (21) was added. The a-celluloses were the residues left after delignification and extraction with 18% (w/v) NaOH of oat straw, birch wood, and ryegrass. 

Dinur T, Osiecki KM, Legler G, Gaft S, Desnick RJ, Grabowski GA (1986) Human add beta-glucosidase isolation and amino acid sequence of a peptide containing the catalytic site. Proc Natl Acad Sci U S A 83 1660-1664... 

Table 5. Properties of p-glucosidases isolated from T. reesei...

An ejco-/3-l,4-D-glucanase (eA u-j8-l,4-D-glucosidase) isolated from almond seeds acted on -D-glucans and jS-D-gluco-oligosaccharides by a mechanism similar to that of glucoamylase. A multiple (two)-subsite model was suggested for the active enzyme-carbohydrate complex on the basis of substrate specificities and rate data. 

His work on enzymes was continued during the Bonn period. Studies on acid phosphatases were carried out that made a major contribution to our knowledge of these enzymes. " The isolation of a crystalline /3-glucosidase from the emulsin of sweet almonds may be regarded as the crowning achievement of his work on glycosidases. ... 

Protein isolation with affinity precipitation has been discussed in detail by Mattiasson and co-workers (see, e.g. Galaev and Mattiassion, 1997) and they have provided an exhaustive tabulation. Polymers varied from alginate.s/chitosan to dextran to NIPAM. Examples of the used proteins are from trypsin, p-glucosidase, xylanase, alkaline protease, etc. It is remarkable that affinity precipitation can sometimes give results comparable to affinity chromatography. 

The sol-gel-entrapped microbial cells have shown excellent tolerance to different alcohols [99], The immobilized E. coli cells followed the Michaelis-Menten equation when quantified with the (3-glucosidase activity via the hydrolysis of 4-nitrophenyl-(3-D-galactopyranosdie [142], The sol-gel matrices doped with gelatin prevented the cell lysis, which usually occurs during the initial gelation process [143], Microorganisms are now widely used in the biosorption of different pollutants and toxicants. Bacillus sphaericus JG-A12 isolated from uranium mining water has been entrapped in aqueous silica nanosol for the accumulation of copper and uranium [144], Premkumar et al. [145] immobilized recombinant luminous bacteria into TEOS sol-gel to study the effect of sol-gel conditions on the cell response (luminescence). The entrapped and free cells showed almost the same intensity of luminescence (little lower), but the entrapped cells were more stable than the free cells (4 weeks at 4°C). This kind of stable cell could be employed in biosensors in the near future. 

Amylo-1 —> 6-glucosidase obtained by Cori and Larner218 from rabbit muscles, and R-enzyme isolated by Hobson, Whelan and Peat219 from potatoes and broad beans, are typical debranching enzymes, which will hydrolyze the 6 — 1-a-D-glucosidic linkage rather than the normal 4 —> 1-a-D linkage. These enzymes will therefore be particularly important in determinations of the fine structure of amylopectin, if they can be sufficiently well purified. 

Castanospermine is a plant alkaloid isolated from the seeds of the Australian chestnut tree, Castanospermium australe. As with most anti-HIV inhibitors, the discovery of the compound predates the time of isolation of HIV-1. The compound inhibits a-glucosidase-1, and therefore normal processing of the glycopro-... 

Isolation yield, using 5 mg (29.5 U) of commercially supplied glucosidase preparation from almond (Sigma, G-0395) per millilitre of reaction mixture. 

A novel 13-membered ring thiocyclitol (214), isolated from Salacia reticulata Wight (family Hippocrateaceae), showed promising a-glucosidase inhibitory activity (IC50 maltase, 0.227 /tM sucrase, 0.186 //M isomaltase, 0.099 however, it did not inhibit a-amylase from porcine pan-... 

The glucosidases involved in the biosynthetic pathway have been studied in detail by Hemscheidt and Zenk 203), and two of the isolated enzymes were specific for the hydrolysis of strictosidine (33) and were purified 120-fold. The enzyme was isolated from a number of indole alkaloid producing plants, including C. roseus, C.pusilus, and C. trichophyllus. The pH optimum was 6.5, and the values were 0.2 mM for enzyme I and 0.1 mM for enzyme II. Molecular weights were estimated at 230,000 for enzyme I and 450,000 for enzyme II. Unlike the case of the enzyme system of Scott et al. 198,199), tryptamine did not activate either of the two enzymes. The enzymes were highly substrate specific vincoside (85)... 

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