Coupling of surface carboxyls of carboxymethylcellulase with aniline via chemical modification

Extreme thermostabilization in aqueous and water-miscible organic mixtures

Saleem A. Bokhari, Ahmed Afzal, M. Hamid Rashid, M. Ibrahim Rajoka, Khawar S. Siddiqui

Research output: Contribution to journalArticle

Abstract

We wish to report the attainment of the highest ever Topt by introducing approximately two aromatic rings through chemical modification of surface carboxyl groups in carboxymethylcellulase from Scopulariopsis sp. with concomitant decrease in Vmax, Km, and optimum pH! This extraordinary enhancement in thermophilicity of aniline-coupled CMCase (Topt = 122°C) by a margin of 73°C as compared with the native enzyme (Topt = 49°C) is the highest reported for any mesophilic enzyme that has been modified either through chemical modification or site-directed mutagenesis. It is also reported for the first time that aniline coupled CMCase (ACC) is simultaneously thermostable in aqueous as well as water-miscible organic solvents. The Topt of native CMCase and ACC were 25 and 90°C, respectively, in 40% (v/v) aqueous dioxan. The modified enzyme was also stabilized against irreversible thermal denaturation. Therefore, at 55°C, ACC had a half-life of 136 min as compared with native CMCase whose half-life was only 5 min. We believe that the reasons for this elevated thermostability and thermophilicity are surface aromatic-aromatic interactions and aromatic interactions with the sugar backbone of the substrate, respectively.

Original languageEnglish (US)
Pages (from-to)276-281
Number of pages6
JournalBiotechnology Progress
Volume18
Issue number2
DOIs
StatePublished - Apr 22 2002

Fingerprint

Water
Half-Life
Enzymes
Scopulariopsis
Site-Directed Mutagenesis
aniline
carboxymethylcellulase
Hot Temperature

ASJC Scopus subject areas

  • Biotechnology

Cite this

Coupling of surface carboxyls of carboxymethylcellulase with aniline via chemical modification : Extreme thermostabilization in aqueous and water-miscible organic mixtures. / Bokhari, Saleem A.; Afzal, Ahmed; Rashid, M. Hamid; Rajoka, M. Ibrahim; Siddiqui, Khawar S.

In: Biotechnology Progress, Vol. 18, No. 2, 22.04.2002, p. 276-281.

Research output: Contribution to journalArticle

@article{54ab62e6b926401cb83f31ceca22712a,
title = "Coupling of surface carboxyls of carboxymethylcellulase with aniline via chemical modification: Extreme thermostabilization in aqueous and water-miscible organic mixtures",
abstract = "We wish to report the attainment of the highest ever Topt by introducing approximately two aromatic rings through chemical modification of surface carboxyl groups in carboxymethylcellulase from Scopulariopsis sp. with concomitant decrease in Vmax, Km, and optimum pH! This extraordinary enhancement in thermophilicity of aniline-coupled CMCase (Topt = 122°C) by a margin of 73°C as compared with the native enzyme (Topt = 49°C) is the highest reported for any mesophilic enzyme that has been modified either through chemical modification or site-directed mutagenesis. It is also reported for the first time that aniline coupled CMCase (ACC) is simultaneously thermostable in aqueous as well as water-miscible organic solvents. The Topt of native CMCase and ACC were 25 and 90°C, respectively, in 40{\%} (v/v) aqueous dioxan. The modified enzyme was also stabilized against irreversible thermal denaturation. Therefore, at 55°C, ACC had a half-life of 136 min as compared with native CMCase whose half-life was only 5 min. We believe that the reasons for this elevated thermostability and thermophilicity are surface aromatic-aromatic interactions and aromatic interactions with the sugar backbone of the substrate, respectively.",
author = "Bokhari, {Saleem A.} and Ahmed Afzal and Rashid, {M. Hamid} and Rajoka, {M. Ibrahim} and Siddiqui, {Khawar S.}",
year = "2002",
month = "4",
day = "22",
doi = "10.1021/bp010146h",
language = "English (US)",
volume = "18",
pages = "276--281",
journal = "Biotechnology Progress",
issn = "8756-7938",
publisher = "John Wiley and Sons Ltd",
number = "2",

}

TY - JOUR

T1 - Coupling of surface carboxyls of carboxymethylcellulase with aniline via chemical modification

T2 - Extreme thermostabilization in aqueous and water-miscible organic mixtures

AU - Bokhari, Saleem A.

AU - Afzal, Ahmed

AU - Rashid, M. Hamid

AU - Rajoka, M. Ibrahim

AU - Siddiqui, Khawar S.

PY - 2002/4/22

Y1 - 2002/4/22

N2 - We wish to report the attainment of the highest ever Topt by introducing approximately two aromatic rings through chemical modification of surface carboxyl groups in carboxymethylcellulase from Scopulariopsis sp. with concomitant decrease in Vmax, Km, and optimum pH! This extraordinary enhancement in thermophilicity of aniline-coupled CMCase (Topt = 122°C) by a margin of 73°C as compared with the native enzyme (Topt = 49°C) is the highest reported for any mesophilic enzyme that has been modified either through chemical modification or site-directed mutagenesis. It is also reported for the first time that aniline coupled CMCase (ACC) is simultaneously thermostable in aqueous as well as water-miscible organic solvents. The Topt of native CMCase and ACC were 25 and 90°C, respectively, in 40% (v/v) aqueous dioxan. The modified enzyme was also stabilized against irreversible thermal denaturation. Therefore, at 55°C, ACC had a half-life of 136 min as compared with native CMCase whose half-life was only 5 min. We believe that the reasons for this elevated thermostability and thermophilicity are surface aromatic-aromatic interactions and aromatic interactions with the sugar backbone of the substrate, respectively.

AB - We wish to report the attainment of the highest ever Topt by introducing approximately two aromatic rings through chemical modification of surface carboxyl groups in carboxymethylcellulase from Scopulariopsis sp. with concomitant decrease in Vmax, Km, and optimum pH! This extraordinary enhancement in thermophilicity of aniline-coupled CMCase (Topt = 122°C) by a margin of 73°C as compared with the native enzyme (Topt = 49°C) is the highest reported for any mesophilic enzyme that has been modified either through chemical modification or site-directed mutagenesis. It is also reported for the first time that aniline coupled CMCase (ACC) is simultaneously thermostable in aqueous as well as water-miscible organic solvents. The Topt of native CMCase and ACC were 25 and 90°C, respectively, in 40% (v/v) aqueous dioxan. The modified enzyme was also stabilized against irreversible thermal denaturation. Therefore, at 55°C, ACC had a half-life of 136 min as compared with native CMCase whose half-life was only 5 min. We believe that the reasons for this elevated thermostability and thermophilicity are surface aromatic-aromatic interactions and aromatic interactions with the sugar backbone of the substrate, respectively.

UR - http://www.scopus.com/inward/record.url?scp=0036010277&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036010277&partnerID=8YFLogxK

U2 - 10.1021/bp010146h

DO - 10.1021/bp010146h

M3 - Article

VL - 18

SP - 276

EP - 281

JO - Biotechnology Progress

JF - Biotechnology Progress

SN - 8756-7938

IS - 2

ER -