Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Study on the interaction between soybean beta-amylase and substrate by the stopped-flow method.

T Kunikata1, H Yamano, T Nagamura

  • 1Laboratory of Biophysical Chemistry, College of Agriculture, University of Osaka Prefecture.

Journal of Biochemistry
|September 1, 1992
PubMed
Summary

Soybean beta-amylase activity was studied using fluorescence to determine enzyme-substrate binding and catalytic rates. Higher binding affinity correlated with smaller fluorescence changes, suggesting active site structural flexibility.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Postoperative delirium is associated with increased long-term mortality following oral and maxillofacial reconstructive surgery.

International journal of oral and maxillofacial surgery·2025
Same author

Decadal stability in coral cover could mask hidden changes on reefs in the East Asian Seas.

Communications biology·2023
Same author

Music before Dental Surgery Suppresses Sympathetic Activity Derived from Preoperative Anxiety: A Randomized Controlled Trial.

JDR clinical and translational research·2019
Same author

Differences in Viscosity of Superior and Inferior Spikelets of Japonica Rice with Various Percentages of Apparent Amylose Content.

Journal of agricultural and food chemistry·2017
Same author

Kinetic Study of the Active Site Structure of β-Amylase from Bacillus cereus var. mycoides.

Bioscience, biotechnology, and biochemistry·2016
Same author

Structural alterations of the superior temporal gyrus in schizophrenia: Detailed subregional differences.

European psychiatry : the journal of the Association of European Psychiatrists·2016

Area of Science:

  • Biochemistry
  • Enzymology

Background:

  • Soybean beta-amylase (EC 3.2.1.2) plays a crucial role in carbohydrate metabolism.
  • Understanding enzyme kinetics and active site dynamics is essential for enzyme engineering.

Purpose of the Study:

  • To investigate the kinetics of soybean beta-amylase hydrolysis of malto-oligosaccharides.
  • To characterize enzyme-substrate and enzyme-product complex formation using fluorescence.
  • To determine molecular activity (k0) and dissociation constants (KdS).

Main Methods:

  • Stopped-flow fluorescence spectroscopy to monitor substrate hydrolysis.
  • Analysis of time-course data to evaluate kinetic parameters.
  • Determination of fluorescence intensity differences (delta F) between free enzyme and enzyme complexes.

Related Experiment Videos

Main Results:

  • Dissociation constants (KdS) and molecular activity (k0) were determined and consistent with previous kinetic data.
  • Fluorescence changes (delta F) for substrates were smaller than for products (G2, G3).
  • A negative correlation between ligand binding affinity and delta F magnitude was observed.

Conclusions:

  • The study provides insights into the binding interactions of soybean beta-amylase with its substrates and products.
  • Smaller delta F values for higher affinity ligands suggest conformational changes in the enzyme's active site upon binding.
  • The findings imply either multiple tryptophan residues in the active site or ligand-induced structural rearrangements.