University of North Florida
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Stuart Chalk, Ph.D.
Department of Chemistry
University of North Florida
Phone: 1-904-620-1938
Fax: 1-904-620-3535
Email: schalk@unf.edu
Website: @unf

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Amyloglucosidase

@ ChemSpider@ NIST@ PubChem

Citations 6

"Simultaneous Determination Of α-amylase And Amyloglucosidase Activities Using Flow Injection Analysis With Fourier Transform Infrared Spectroscopic Detection And Partial Least-squares Data Treatment"
Anal. Chim. Acta 1998 Volume 366, Issue 1-3 Pages 35-43
R. Schindler, B. Lendl* and R. Kellner

Abstract: A flow injection analysis (FIA) system with fourier transform IR (FTIR) spectroscopic detection for the simultaneous determination of amyloglucosidase (exo-glucan 1,4-α-glucohydrolase, EC 3.2.1.3) and α-amylase (endo-glucan 1,4-α-glucanohydrolase, EC 3.2.1.1) activities in aqueous solution is proposed. Starch hydrolysis catalyzed by both enzymes was monitored in the mid-IR range by recording IR-spectra of the assay solutions before and after hydrolysis. The intensities of the resulting difference spectra were directly related to the enzyme activities. Furthermore, the different reaction pathways of the two enzymes under investigation produced distinct spectral changes resulting in difference spectra characteristic for each enzyme. Using partial least squares (PLS) regression it was possible to simultaneously determine both enzyme activities covering a range of 38-150 U/l (630-2500 nkat/l) for amyloglucosidase and a range of 500-3750 U/l (8300-62000 nkat/l) for α-amylase. Absorption due to the sample matrix was successfully eliminated by calculation of the difference spectra. The developed method was also successfully applied to the anal. of spiked fermentation broth samples yielding mean deviations of 2.6% and 10.8% for amyloglucosidase and α-amylase, respectively.
Fermentation broth Spectrophotometry Simultaneous analysis Catalysis Interferences Partial least squares

"Multistep Determination Of Enzyme Activity By Flow Injection And Sequential-injection Analysis. Assay Of Amyloglucosidase"
Talanta 1994 Volume 41, Issue 11 Pages 1881-1893
Elo Harald Hansen, Bodil Willumsen, Solveig K. Winther and Helle Drabøl,

Abstract: A reactor vessel containing 27.8 mM maltose substrate in 5 mM acetate buffer of pH 4.3 was heated to 37°C and amyloglucosidase (glucan 1,4-α-glucosidase) was added with stirring. Samples were withdrawn at preset times into a previously described FIA system (Hansen and Jensen, Ibid., 1993, 40, 1891) for spectrophotometric detection. Glucan 1,4-α-glucoside was also assayed using a sequential injection analysis (SIA) system. The SIA system comprised a peristaltic pump, a 10 port electrically actuated multi-position directional valve and a computer. The water carrier solution, 50 g/l maltose solution in 0.02 M acetate, sample solution and glucose dehydrogenase reagent were placed around the valve along side a narrow glass vessel mixing chamber with a stirring bar in a thermostatted water bath at 37°C. The flow rate was 1.2 ml/min and detection was by diode-array spectrophotometry. Both FIA and SIA allowed two mutually incompatible reactions to be completely separated.
Biological Spectrophotometry Sequential injection Heated reaction Valve Well stirred mixing chamber

"Automatic Spectrophotometric Determination Of Amyloglucosidase Activity Using P-nitrophenyl-α-D Glucopyranoside And A Flow Injection Analyzer"
Analyst 1986 Volume 111, Issue 8 Pages 927-929
Kaj André Holm

Abstract: The method is based on the hydrolysis of 4-nitrophenyl-α-D-glucopyranoside(I) by the cited enzyme, glucan 1,4-α-glucosidase(II), and spectrophotometric determination of the 4-nitrophenol released. The sample (30 µL) is injected into a water carrier stream (1.4 mL min-1), which is mixed with a reagent stream (0.53 mL min-1) containing 2 g L-1 of I in 0.1 M acetate buffer (pH 4.3). The solution is incubated for 20 s at 50°C, passed through a reaction coil at 60°C, and mixed with 0.1 M Na2CO3 (0.53 mL min-1) before the absorbance is measured at 400 nm. Similar procedures have been developed for the determination of α-L-arabinofuranosidase and α-galactosidase activity with 4-nitrophenyl-α-L-arabinofuranoside and 4-nitrophenyl-α-D-galactopyranoside as chromophores, respectively.
Spectrophotometry Enzyme Heated reaction

"Determination Of Amyloglucosidase Activity Using Flow Injection Analysis With Fourier Transform Infrared Spectrometric Detection"
Analyst 1997 Volume 122, Issue 6 Pages 531-534
R. Schindler, B. Lendl and R. Kellner

Abstract: Samples (250 µL each) and a 55 g/l starch solution in 0.1 M acetate buffer of pH 4.3 were simultaneously injected into two aqueous carrier streams (both at a flow rate of 0.88 ml/min) and merged in a reaction coil (250 cm x 0.5 mm i.d.) maintained at 54°C. When the reaction plug had filled the reaction coil, the flow was stopped for 5 min after which the FTIR spectrum was recorded from 950-1300 cm-1. The amyloglucosidase (I) activity was calculated from the difference in the absorbances at 1078 and 1020 cm-1 compared with a reference spectrum of unreacted starch. The calibration graph was linear from 50-2000 U/l I. The method was applied to fermentation broths: recoveries were 98-102% of added I.
Fermentation broth Spectrophotometry Stopped-flow Heated reaction

"A Comparative Kinetic Study Of Soluble And Immobilized Amyloglucosidase On Non-porous Glass Beads With An Automated Flow Injection Analysis System"
Acta Chim. Hung. 1992 Volume 129, Issue 3-4 Pages 461-468
Kiranas, E.R.;Tzouwara Karayanni, S.M.;Karayannis, M.I.

Abstract: A comparative kinetic study of solution and immobilized amyloglucosidase (AG; EC 3.2.1.3) on non-porous glass beads is presented with an automated flow injection analysis (FIA) system, with maltose as substrate. All measurements were based on the catalytic action of the enzymes soluble or immobilized AG, immobilized glucose oxidase, and soluble horseradish peroxidase. The main results are: (i) optimization of the attachment time of glutaraldehyde during the immobilization procedure; (ii) a detailed study on the pH of immobilization to find its optimum value for enzyme activity; (iii) determination of the half-life of AG single-bead string reactors under continuous operational conditions; (iv) determination of the kinetic parameters (Km and Amax) of the soluble and immobilized AG with maltose as substrate and study of the aging effects of immobilized AG on the kinetic parameters; (v) development of an analytical FIA mode, for the determination of maltose.
Immobilized enzyme Glass beads Optimization Kinetic Single bead string reactor

"Flow Injection Analysis For Rapid Amyloglucosidase Activity Determination"
J. Food Comp. Anal. 1989 Volume 2, Issue 4 Pages 364-370
Clark G. Hartford, David W. Muntz, James V. Evans and Gary T. Blair

Abstract: Modifications were made on the procedure described by Holm (Anal. Abstr., 1987, 49, 5D367) for the determination of glucan 1,4-α-glucosidase (I) which involved an increased sampling rate, replacement of Na2CO3 with sodium borate buffer for a better pH adjustment and elimination of the stopped-flow procedure. The calibration graph was essentially linear between 0.05 and 2.5 diazyme unit (DU) mL-1 (r 0.9996). The sensitivity was 0.1 DU g-1 although I levels in most samples were from 40 to 400 DUu g-1. The coefficient of variation was 1.2% compared with 3.9% for a manual method.
Buffer Calibration Sensitivity Stopped-flow