The use of starch azure for measurement of alpha-amylase activity

Gábor Lehoczki, L. Kandra, G. Gyémánt

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Despite being widely used, there is no standard protocol for α-amylase activity measurement with starch azure substrate. Boiling pre-treatment of starch azure suspension increased the reaction rate of hydrolysis catalysed by human salivary α-amylase (HSA) or porcine pancreatic α-amylase (PPA) and the sensitivity of spectrophotometric activity measurement has been improved. Kinetic constants, KM, and vmax, obtained from parallel isothermal titration calorimetric (ITC) measurements on natural and starch azure revealed, that the blue starch derivative does not differ significantly from its natural counterpart from kinetic point of view. Interestingly, substrate inhibition was observed in starch azure hydrolysis characterised by dissociation constants 49 mg/mL and 16.4 mg/mL for HSA and PPA, respectively. In this work a new protocol has been suggested for α-amylase activity measurement using boiled insoluble starch azure as substrate at 5 mg/mL concentration.

Original languageEnglish
Pages (from-to)263-266
Number of pages4
JournalCarbohydrate Polymers
Volume183
DOIs
Publication statusPublished - Mar 1 2018

Fingerprint

Amylases
alpha-Amylases
Starch
Hydrolysis
Substrates
Kinetics
Titration
Boiling liquids
Reaction rates
Suspensions
Derivatives

Keywords

  • Amylase
  • Assay method
  • Microcalorimetry
  • Starch azure

ASJC Scopus subject areas

  • Organic Chemistry
  • Polymers and Plastics
  • Materials Chemistry

Cite this

The use of starch azure for measurement of alpha-amylase activity. / Lehoczki, Gábor; Kandra, L.; Gyémánt, G.

In: Carbohydrate Polymers, Vol. 183, 01.03.2018, p. 263-266.

Research output: Contribution to journalArticle

@article{a062b19fb44e4d70a38c122abd9eb4f0,
title = "The use of starch azure for measurement of alpha-amylase activity",
abstract = "Despite being widely used, there is no standard protocol for α-amylase activity measurement with starch azure substrate. Boiling pre-treatment of starch azure suspension increased the reaction rate of hydrolysis catalysed by human salivary α-amylase (HSA) or porcine pancreatic α-amylase (PPA) and the sensitivity of spectrophotometric activity measurement has been improved. Kinetic constants, KM, and vmax, obtained from parallel isothermal titration calorimetric (ITC) measurements on natural and starch azure revealed, that the blue starch derivative does not differ significantly from its natural counterpart from kinetic point of view. Interestingly, substrate inhibition was observed in starch azure hydrolysis characterised by dissociation constants 49 mg/mL and 16.4 mg/mL for HSA and PPA, respectively. In this work a new protocol has been suggested for α-amylase activity measurement using boiled insoluble starch azure as substrate at 5 mg/mL concentration.",
keywords = "Amylase, Assay method, Microcalorimetry, Starch azure",
author = "G{\'a}bor Lehoczki and L. Kandra and G. Gy{\'e}m{\'a}nt",
year = "2018",
month = "3",
day = "1",
doi = "10.1016/j.carbpol.2017.12.037",
language = "English",
volume = "183",
pages = "263--266",
journal = "Carbohydrate Polymers",
issn = "0144-8617",
publisher = "Elsevier Limited",

}

TY - JOUR

T1 - The use of starch azure for measurement of alpha-amylase activity

AU - Lehoczki, Gábor

AU - Kandra, L.

AU - Gyémánt, G.

PY - 2018/3/1

Y1 - 2018/3/1

N2 - Despite being widely used, there is no standard protocol for α-amylase activity measurement with starch azure substrate. Boiling pre-treatment of starch azure suspension increased the reaction rate of hydrolysis catalysed by human salivary α-amylase (HSA) or porcine pancreatic α-amylase (PPA) and the sensitivity of spectrophotometric activity measurement has been improved. Kinetic constants, KM, and vmax, obtained from parallel isothermal titration calorimetric (ITC) measurements on natural and starch azure revealed, that the blue starch derivative does not differ significantly from its natural counterpart from kinetic point of view. Interestingly, substrate inhibition was observed in starch azure hydrolysis characterised by dissociation constants 49 mg/mL and 16.4 mg/mL for HSA and PPA, respectively. In this work a new protocol has been suggested for α-amylase activity measurement using boiled insoluble starch azure as substrate at 5 mg/mL concentration.

AB - Despite being widely used, there is no standard protocol for α-amylase activity measurement with starch azure substrate. Boiling pre-treatment of starch azure suspension increased the reaction rate of hydrolysis catalysed by human salivary α-amylase (HSA) or porcine pancreatic α-amylase (PPA) and the sensitivity of spectrophotometric activity measurement has been improved. Kinetic constants, KM, and vmax, obtained from parallel isothermal titration calorimetric (ITC) measurements on natural and starch azure revealed, that the blue starch derivative does not differ significantly from its natural counterpart from kinetic point of view. Interestingly, substrate inhibition was observed in starch azure hydrolysis characterised by dissociation constants 49 mg/mL and 16.4 mg/mL for HSA and PPA, respectively. In this work a new protocol has been suggested for α-amylase activity measurement using boiled insoluble starch azure as substrate at 5 mg/mL concentration.

KW - Amylase

KW - Assay method

KW - Microcalorimetry

KW - Starch azure

UR - http://www.scopus.com/inward/record.url?scp=85038811034&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85038811034&partnerID=8YFLogxK

U2 - 10.1016/j.carbpol.2017.12.037

DO - 10.1016/j.carbpol.2017.12.037

M3 - Article

C2 - 29352883

AN - SCOPUS:85038811034

VL - 183

SP - 263

EP - 266

JO - Carbohydrate Polymers

JF - Carbohydrate Polymers

SN - 0144-8617

ER -