Characterisation of recombinant Trichoderma reesei cellobiohydrolase and the potential of cellulase mixture in hydrolyzing oil palm empty fruit bunches

Doris Quay Huai Xia, Yong Hui Yee, Rosli Md Illias, Nor Muhammad Mahadi, Farah Diba Abu Bakar, Abdul Munir Abd. Murad

Research output: Contribution to journalArticle

Abstract

The potential for recombinant cellulases from fungi to hydrolyze oil palm empty fruit bunches (OPEFB) into simple sugars was investigated. A mixture of two recombinant enzymes consisting of a cellobiohydrolase from Trichoderma reesei and an endoglucanase from Aspergillus niger, was evaluated for OPEFB hydrolysis. The development of A. niger endoglucanase has been described previously. In this work, the development of a recombinant cellobiohydrolase and the activity of this enzyme mixture towards OPEFB hydrolysis were described. To obtain recombinant cellobiohydrolase, the cDNA encoding for cellobiohydrolase, cbhII, was isolated from the T. reesei strain M5, and the gene was expressed in the methylotrophic yeast, Pichia pastoris. Partially purified CbhII demonstrated optimum activity at 50°C and pH 5.0. This enzyme was shown to hydrolyze Avicel at a concentration of 0.486 U/mg under optimum conditions. Hydrolysis of pretreated OPEFB using CbhII and endoglucanase EglA showed that the total reducing sugar produced was higher in the enzyme mixture reaction than were produced in an individual enzyme reaction. Addition of β-glucosidase and an enzyme cofactor significantly increased enzyme activity and OPEFB cellulose hydrolysis. This study demonstrated the capability of an enzyme mixture to produce sugars from oil palm lignocellulosic waste.

Original languageEnglish
Pages (from-to)11-19
Number of pages9
JournalMalaysian Applied Biology
Volume46
Issue number2
Publication statusPublished - 2017

Fingerprint

Cellulose 1,4-beta-Cellobiosidase
cellulose 1,4-beta-cellobiosidase
Trichoderma reesei
Trichoderma
Cellulase
Elaeis guineensis
endo-1,4-beta-glucanase
Fruit
Oils
fruits
Enzymes
hydrolysis
Hydrolysis
enzymes
Aspergillus niger
Cellulose
lignocellulosic wastes
enzyme activity
sugars
Pichia pastoris

Keywords

  • Cellobiohydrolase
  • Enzyme mixture
  • Oil palm empty fruit bunch
  • Trichoderma reesei

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)

Cite this

Characterisation of recombinant Trichoderma reesei cellobiohydrolase and the potential of cellulase mixture in hydrolyzing oil palm empty fruit bunches. / Xia, Doris Quay Huai; Yee, Yong Hui; Illias, Rosli Md; Mahadi, Nor Muhammad; Abu Bakar, Farah Diba; Abd. Murad, Abdul Munir.

In: Malaysian Applied Biology, Vol. 46, No. 2, 2017, p. 11-19.

Research output: Contribution to journalArticle

@article{df81095ec6c74185834a610267a5f760,
title = "Characterisation of recombinant Trichoderma reesei cellobiohydrolase and the potential of cellulase mixture in hydrolyzing oil palm empty fruit bunches",
abstract = "The potential for recombinant cellulases from fungi to hydrolyze oil palm empty fruit bunches (OPEFB) into simple sugars was investigated. A mixture of two recombinant enzymes consisting of a cellobiohydrolase from Trichoderma reesei and an endoglucanase from Aspergillus niger, was evaluated for OPEFB hydrolysis. The development of A. niger endoglucanase has been described previously. In this work, the development of a recombinant cellobiohydrolase and the activity of this enzyme mixture towards OPEFB hydrolysis were described. To obtain recombinant cellobiohydrolase, the cDNA encoding for cellobiohydrolase, cbhII, was isolated from the T. reesei strain M5, and the gene was expressed in the methylotrophic yeast, Pichia pastoris. Partially purified CbhII demonstrated optimum activity at 50°C and pH 5.0. This enzyme was shown to hydrolyze Avicel at a concentration of 0.486 U/mg under optimum conditions. Hydrolysis of pretreated OPEFB using CbhII and endoglucanase EglA showed that the total reducing sugar produced was higher in the enzyme mixture reaction than were produced in an individual enzyme reaction. Addition of β-glucosidase and an enzyme cofactor significantly increased enzyme activity and OPEFB cellulose hydrolysis. This study demonstrated the capability of an enzyme mixture to produce sugars from oil palm lignocellulosic waste.",
keywords = "Cellobiohydrolase, Enzyme mixture, Oil palm empty fruit bunch, Trichoderma reesei",
author = "Xia, {Doris Quay Huai} and Yee, {Yong Hui} and Illias, {Rosli Md} and Mahadi, {Nor Muhammad} and {Abu Bakar}, {Farah Diba} and {Abd. Murad}, {Abdul Munir}",
year = "2017",
language = "English",
volume = "46",
pages = "11--19",
journal = "Malaysian Applied Biology",
issn = "0126-8643",
publisher = "Malaysian Society of Applied Biology",
number = "2",

}

TY - JOUR

T1 - Characterisation of recombinant Trichoderma reesei cellobiohydrolase and the potential of cellulase mixture in hydrolyzing oil palm empty fruit bunches

AU - Xia, Doris Quay Huai

AU - Yee, Yong Hui

AU - Illias, Rosli Md

AU - Mahadi, Nor Muhammad

AU - Abu Bakar, Farah Diba

AU - Abd. Murad, Abdul Munir

PY - 2017

Y1 - 2017

N2 - The potential for recombinant cellulases from fungi to hydrolyze oil palm empty fruit bunches (OPEFB) into simple sugars was investigated. A mixture of two recombinant enzymes consisting of a cellobiohydrolase from Trichoderma reesei and an endoglucanase from Aspergillus niger, was evaluated for OPEFB hydrolysis. The development of A. niger endoglucanase has been described previously. In this work, the development of a recombinant cellobiohydrolase and the activity of this enzyme mixture towards OPEFB hydrolysis were described. To obtain recombinant cellobiohydrolase, the cDNA encoding for cellobiohydrolase, cbhII, was isolated from the T. reesei strain M5, and the gene was expressed in the methylotrophic yeast, Pichia pastoris. Partially purified CbhII demonstrated optimum activity at 50°C and pH 5.0. This enzyme was shown to hydrolyze Avicel at a concentration of 0.486 U/mg under optimum conditions. Hydrolysis of pretreated OPEFB using CbhII and endoglucanase EglA showed that the total reducing sugar produced was higher in the enzyme mixture reaction than were produced in an individual enzyme reaction. Addition of β-glucosidase and an enzyme cofactor significantly increased enzyme activity and OPEFB cellulose hydrolysis. This study demonstrated the capability of an enzyme mixture to produce sugars from oil palm lignocellulosic waste.

AB - The potential for recombinant cellulases from fungi to hydrolyze oil palm empty fruit bunches (OPEFB) into simple sugars was investigated. A mixture of two recombinant enzymes consisting of a cellobiohydrolase from Trichoderma reesei and an endoglucanase from Aspergillus niger, was evaluated for OPEFB hydrolysis. The development of A. niger endoglucanase has been described previously. In this work, the development of a recombinant cellobiohydrolase and the activity of this enzyme mixture towards OPEFB hydrolysis were described. To obtain recombinant cellobiohydrolase, the cDNA encoding for cellobiohydrolase, cbhII, was isolated from the T. reesei strain M5, and the gene was expressed in the methylotrophic yeast, Pichia pastoris. Partially purified CbhII demonstrated optimum activity at 50°C and pH 5.0. This enzyme was shown to hydrolyze Avicel at a concentration of 0.486 U/mg under optimum conditions. Hydrolysis of pretreated OPEFB using CbhII and endoglucanase EglA showed that the total reducing sugar produced was higher in the enzyme mixture reaction than were produced in an individual enzyme reaction. Addition of β-glucosidase and an enzyme cofactor significantly increased enzyme activity and OPEFB cellulose hydrolysis. This study demonstrated the capability of an enzyme mixture to produce sugars from oil palm lignocellulosic waste.

KW - Cellobiohydrolase

KW - Enzyme mixture

KW - Oil palm empty fruit bunch

KW - Trichoderma reesei

UR - http://www.scopus.com/inward/record.url?scp=85021765350&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85021765350&partnerID=8YFLogxK

M3 - Article

VL - 46

SP - 11

EP - 19

JO - Malaysian Applied Biology

JF - Malaysian Applied Biology

SN - 0126-8643

IS - 2

ER -