Two simple and rapid methods for the detection of polymer-degrading enzymes on high-resolution, alkaline, cold, in situ-native (HiRACIN)-PAGE and high-resolution, in situ-inhibited native (HiRISIN)-PAGE

Ahmed Afzal, Salim Ahmed Bokhari, Waseem Ahmad, Mohammad Hamid Rashid, Mohammad Ibrahim Rajoka, Khawar Sohail Siddiqui

Research output: Contribution to journalArticle

Abstract

Two sensitive, high-resolution and exceedingly versatile methods for the detection of isoenzymes of polymer-degrading enzymes on high-resolution, alkaline, cold, in situ-native (HiRACIN)-PAGE and high-resolution in situ- inhibited, native (HiRISIN)-PAGE are described. Extracellular crude extracts containing xylanases and carboxymethylcellulases from Scopulariopsis sp. and glucoamylases from Aspergillus niger were subjected to non-denaturing PAGE containing substrates in the resolving gel. In case of HiRACIN-PAGE, the enzymes were prevented from degrading their respective substrates during run by carrying out electrophoresis at 4 °C and the pH of running and resolving gel buffer systems were increased from 8.5 to 10.6. In case of HiRISIN-PAGE, adding competitive inhibitor of the enzyme, cellobiose, in the resolving gel prevents the degradation of polymer during the run. These techniques were successfully applied, for the first time, to visualize four, three and four sharp and distinct bands of xylanases, glucoamylases and CMCases, respectively.

Original languageEnglish (US)
Pages (from-to)957-960
Number of pages4
JournalBiotechnology Letters
Volume22
Issue number11
DOIs
StatePublished - Jan 1 2000

Fingerprint

Native Polyacrylamide Gel Electrophoresis
Glucan 1,4-alpha-Glucosidase
Polymers
Gels
Enzymes
Isoenzymes
Cellobiose
Aspergillus
Enzyme Inhibitors
Substrates
Electrophoresis
Complex Mixtures
Scopulariopsis
Buffers
Aspergillus niger
Degradation

Keywords

  • Electrophoresis
  • Endoglucanase
  • Glucoamylase
  • Xylanase

ASJC Scopus subject areas

  • Biotechnology

Cite this

Two simple and rapid methods for the detection of polymer-degrading enzymes on high-resolution, alkaline, cold, in situ-native (HiRACIN)-PAGE and high-resolution, in situ-inhibited native (HiRISIN)-PAGE. / Afzal, Ahmed; Bokhari, Salim Ahmed; Ahmad, Waseem; Rashid, Mohammad Hamid; Rajoka, Mohammad Ibrahim; Siddiqui, Khawar Sohail.

In: Biotechnology Letters, Vol. 22, No. 11, 01.01.2000, p. 957-960.

Research output: Contribution to journalArticle

Afzal, Ahmed ; Bokhari, Salim Ahmed ; Ahmad, Waseem ; Rashid, Mohammad Hamid ; Rajoka, Mohammad Ibrahim ; Siddiqui, Khawar Sohail. / Two simple and rapid methods for the detection of polymer-degrading enzymes on high-resolution, alkaline, cold, in situ-native (HiRACIN)-PAGE and high-resolution, in situ-inhibited native (HiRISIN)-PAGE. In: Biotechnology Letters. 2000 ; Vol. 22, No. 11. pp. 957-960.
@article{ed6ff2c7e71f465fb838349750a72cc6,
title = "Two simple and rapid methods for the detection of polymer-degrading enzymes on high-resolution, alkaline, cold, in situ-native (HiRACIN)-PAGE and high-resolution, in situ-inhibited native (HiRISIN)-PAGE",
abstract = "Two sensitive, high-resolution and exceedingly versatile methods for the detection of isoenzymes of polymer-degrading enzymes on high-resolution, alkaline, cold, in situ-native (HiRACIN)-PAGE and high-resolution in situ- inhibited, native (HiRISIN)-PAGE are described. Extracellular crude extracts containing xylanases and carboxymethylcellulases from Scopulariopsis sp. and glucoamylases from Aspergillus niger were subjected to non-denaturing PAGE containing substrates in the resolving gel. In case of HiRACIN-PAGE, the enzymes were prevented from degrading their respective substrates during run by carrying out electrophoresis at 4 °C and the pH of running and resolving gel buffer systems were increased from 8.5 to 10.6. In case of HiRISIN-PAGE, adding competitive inhibitor of the enzyme, cellobiose, in the resolving gel prevents the degradation of polymer during the run. These techniques were successfully applied, for the first time, to visualize four, three and four sharp and distinct bands of xylanases, glucoamylases and CMCases, respectively.",
keywords = "Electrophoresis, Endoglucanase, Glucoamylase, Xylanase",
author = "Ahmed Afzal and Bokhari, {Salim Ahmed} and Waseem Ahmad and Rashid, {Mohammad Hamid} and Rajoka, {Mohammad Ibrahim} and Siddiqui, {Khawar Sohail}",
year = "2000",
month = "1",
day = "1",
doi = "10.1023/A:1005676824462",
language = "English (US)",
volume = "22",
pages = "957--960",
journal = "Biotechnology Letters",
issn = "0141-5492",
publisher = "Springer Netherlands",
number = "11",

}

TY - JOUR

T1 - Two simple and rapid methods for the detection of polymer-degrading enzymes on high-resolution, alkaline, cold, in situ-native (HiRACIN)-PAGE and high-resolution, in situ-inhibited native (HiRISIN)-PAGE

AU - Afzal, Ahmed

AU - Bokhari, Salim Ahmed

AU - Ahmad, Waseem

AU - Rashid, Mohammad Hamid

AU - Rajoka, Mohammad Ibrahim

AU - Siddiqui, Khawar Sohail

PY - 2000/1/1

Y1 - 2000/1/1

N2 - Two sensitive, high-resolution and exceedingly versatile methods for the detection of isoenzymes of polymer-degrading enzymes on high-resolution, alkaline, cold, in situ-native (HiRACIN)-PAGE and high-resolution in situ- inhibited, native (HiRISIN)-PAGE are described. Extracellular crude extracts containing xylanases and carboxymethylcellulases from Scopulariopsis sp. and glucoamylases from Aspergillus niger were subjected to non-denaturing PAGE containing substrates in the resolving gel. In case of HiRACIN-PAGE, the enzymes were prevented from degrading their respective substrates during run by carrying out electrophoresis at 4 °C and the pH of running and resolving gel buffer systems were increased from 8.5 to 10.6. In case of HiRISIN-PAGE, adding competitive inhibitor of the enzyme, cellobiose, in the resolving gel prevents the degradation of polymer during the run. These techniques were successfully applied, for the first time, to visualize four, three and four sharp and distinct bands of xylanases, glucoamylases and CMCases, respectively.

AB - Two sensitive, high-resolution and exceedingly versatile methods for the detection of isoenzymes of polymer-degrading enzymes on high-resolution, alkaline, cold, in situ-native (HiRACIN)-PAGE and high-resolution in situ- inhibited, native (HiRISIN)-PAGE are described. Extracellular crude extracts containing xylanases and carboxymethylcellulases from Scopulariopsis sp. and glucoamylases from Aspergillus niger were subjected to non-denaturing PAGE containing substrates in the resolving gel. In case of HiRACIN-PAGE, the enzymes were prevented from degrading their respective substrates during run by carrying out electrophoresis at 4 °C and the pH of running and resolving gel buffer systems were increased from 8.5 to 10.6. In case of HiRISIN-PAGE, adding competitive inhibitor of the enzyme, cellobiose, in the resolving gel prevents the degradation of polymer during the run. These techniques were successfully applied, for the first time, to visualize four, three and four sharp and distinct bands of xylanases, glucoamylases and CMCases, respectively.

KW - Electrophoresis

KW - Endoglucanase

KW - Glucoamylase

KW - Xylanase

UR - http://www.scopus.com/inward/record.url?scp=0033933337&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033933337&partnerID=8YFLogxK

U2 - 10.1023/A:1005676824462

DO - 10.1023/A:1005676824462

M3 - Article

AN - SCOPUS:0033933337

VL - 22

SP - 957

EP - 960

JO - Biotechnology Letters

JF - Biotechnology Letters

SN - 0141-5492

IS - 11

ER -