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Volumn 39, Issue 19 SUPPL., 2000, Pages 177-180

Direct NMR-spectroscopic determination of active-enzyme concentration by titration with a labeled inhibitor: Determination of the kcat value of almond β-glucosidase

Author keywords

Azasugars; Enzyme inhibitors; Enzyme kinetics; Glycosidases; NMR spectroscopy

Indexed keywords

1 AZAFAGOMINE; 1-AZAFAGOMINE; BETA GLUCOSIDASE; ENZYME INHIBITOR; GLYCOSIDE; INDOLIZINE DERIVATIVE;

EID: 0039172825     PISSN: 05700833     EISSN: None     Source Type: Journal    
DOI: None     Document Type: Article
Times cited : (9)

References (18)
  • 1
    • 0001441618 scopus 로고
    • F. W. Lichtenthaler, Angew. Chem. 1994, 106, 2456-2467; Angew. Chem. Int. Ed. Engl. 1994, 33, 2364-2374.
    • (1994) Angew. Chem. , vol.106 , pp. 2456-2467
    • Lichtenthaler, F.W.1
  • 2
    • 0001441617 scopus 로고
    • F. W. Lichtenthaler, Angew. Chem. 1994, 106, 2456-2467; Angew. Chem. Int. Ed. Engl. 1994, 33, 2364-2374.
    • (1994) Angew. Chem. Int. Ed. Engl. , vol.33 , pp. 2364-2374
  • 4
    • 0040040756 scopus 로고    scopus 로고
    • note
    • In the context of this work, specific binding is defined as binding of the inhibitor to the active site of the enzyme. Unspecific binding is defined as adsorption of the inhibitor on the enzyme in a random way.
  • 8
    • 0040634076 scopus 로고    scopus 로고
    • note
    • Spectra were generally run without internal reference, but vertical scaling factors were used. The peak height of the sample signal was compared with the peak height of a standard signal determined at the vertical scaling. The method was tested repeatedly and was reliable for determining the inhibitor concentration. Experiments with dioxane as an internal reference were also carried out, but it was found that dioxane bound to the enzyme unspecifically (to a higher degree than the inhibitor). Presence of the reference thus only led to complications.
  • 11
    • 0003751067 scopus 로고    scopus 로고
    • Worthington Biochemical Corporation, Lakewood, NJ, USA
    • Worthington Enzyme Manual, Worthington Biochemical Corporation, Lakewood, NJ, USA (http://www.worthington-biochem.com/manual/manIndex.html).
    • Worthington Enzyme Manual
  • 14
    • 0038855880 scopus 로고    scopus 로고
    • note
    • One unit of α-glucosidase, β-glucosidase, and isomaltase hydrolyze, per definition (Sigma), 1 μmol of p-nitrophenyl-α-D-glycoside per minute, while one unit of glucoamylase hydrolyze 1 μmol of maltose per minute, both at 37°C.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.