Functional screening of metagenomic libraries is an effective approach for identification novel enzymes. A Caatinga biome goat rumen library was screened using esculin as a substrate, and gene from unknown bacterium encoding GH3 enzyme, BGL11, identified. None the BGL11 closely related genes have been previously characterized. Recombinant obtained kinetically Substrate specificity purified prot...

The uncharacterized protein Rsp3690 from Rhodobacter sphaeroides is a member of the amidohydrolase superfamily of enzymes. In this investigation the gene for Rsp3690 was expressed in Escherichia coli and purified to homogeneity, and the three-dimensional structure was determined to a resolution of 1.8 Å. The protein folds as a distorted (β/α)8-barrel, and the subunits associate as a homotetrame...

( 1) A method is described for assaying brain for cerebroside galactosidase activity. The enzyme was liberated by sonication and addition of sodium taurocholate, then by digestion with pancreatic enzymes. It was further purified by precipitation at pH 3. The enzyme was then incubated with an emulsion of galactose-labelled cerebroside in taurocholate and oleate at pH 4.5, and the liberated galac...

Comparative reactivities of three diaqua(tetraamine)Co(III) complexes towards hydrolysis of 4-nitrophenyl phosphate was undertaken. The results were contrasted with prussian blue promoted hydrolysis of 4-nitrophenyl phosphate. The tetraamineCobalt(III) complexes accelerated the hydrolytic reaction to varying degrees with the following reactivity order. [(tn)2Co(OH2)2] < [(trpn)Co(OH2)2] < [(tme...

Several esters of 4-methylumbelliferone and 2-naphthol were synthesized and examined as possible spectrofluorimetric titrants for bovine alpha-chymotrypsin, trypsin, thrombin, Factor Xa and for subtilisin Novo. 4-Methylumbelliferyl p-guanidinobenzoate hydrochloride (MUGB) is a satisfactory titrant for alpha- and beta-trypsin, thrombin and Factor Xa and 4-methylumbelliferyl p-(NNN-trimethylammon...

With p-nitrophenyl phosphate as the substrate, there reportedly is no organ-specific inhibition of alkaline phosphatase (EC 3.1.3.1) activity by L-phenylalanine. However, we found that at pH 10.0, with p-nitrophenyl phosphate as the substrate, L-phenylalanine obviously inhibits the alkaline phosphatase isoenzyme from human placenta, whereas there is little if any inhibition of the isoenzyme fro...

p-Nitrophenyl-p’-guanidinobenzoate is a suitable substrate for the titration of purified bovine Factor X which has been activated by a coagulant protein from Russell’s viper venom. Reaction of Factor Xa with p-nitrophenyl-P’-guanidinobenzoate occurs at the site which is responsible for the clotting activity of Factor Xa. Activation of bovine Factor X by the Russell’s viper venom coagulant prote...

p-Nitrophenyl-p’-guanidinobenzoate is a suitable substrate for the titration of purified bovine Factor X which has been activated by a coagulant protein from Russell’s viper venom. Reaction of Factor Xa with p-nitrophenyl-P’-guanidinobenzoate occurs at the site which is responsible for the clotting activity of Factor Xa. Activation of bovine Factor X by the Russell’s viper venom coagulant prote...

A cytosolic phosphoprotein phosphatase of M, = 95,000 purified from bovine cardiac muscle, which contains a catalytic subunit of M, = 35,000, is known to be associated with a Mg2+-activated p-nitrophenyl phosphatase activity. We have found that the enzyme preparation is also active toward phosphotyrosyl-IgG and -casein phosphorylated by pp60"", the transforming gene product of Rous sarcoma viru...