Protein engineering of GH11 xylanase from Aspergillus fumigatus RT-1 for catalytic efficiency improvement on kenaf biomass hydrolysis

Siti Intan Rosdianah Damis, Abdul Munir Abd. Murad, Farah Diba Abu Bakar, Siti Aishah Rashid, Nardiah Rizwana Jaafar, Rosli Md Illias

Research output: Contribution to journalArticle

Abstract

Enzyme hydrolysis faces a bottleneck due to the recalcitrance of the lignocellulose biomass. The protein engineering of GH11 xylanase from Aspergillus fumigatus RT-1 was performed near the active site and at the N-terminal region to improve its catalytic efficiency towards pretreated kenaf (Hibiscus cannabinus) hydrolysis. Five mutants were constructed by combined approaches of error-prone PCR, site-saturation and site-directed mutagenesis. The double mutant c168 t/Q192H showed the most effective hydrolysis reaction with a 13.9-fold increase in catalytic efficiency, followed by mutants Y7L and c168 t/Q192 H/Y7L with a 1.6-fold increase, respectively. The enhanced catalytic efficiency evoked an increase in sugar yield of up to 28% from pretreated kenaf. In addition, mutant c168 t/Q192 H/Y7L improved the thermostability at higher temperature and acid stability. This finding shows that mutations at distances less than 15 Å from the active site and at putative secondary binding sites affect xylanase catalytic efficiency towards insoluble substrates hydrolysis.

Original languageEnglish
Article number109383
JournalEnzyme and Microbial Technology
Volume131
DOIs
Publication statusPublished - 1 Dec 2019

Fingerprint

Hibiscus
Protein Engineering
Aspergillus fumigatus
Aspergillus
Biomass
Hydrolysis
Proteins
Catalytic Domain
Mutagenesis
Binding sites
Site-Directed Mutagenesis
Sugars
Enzymes
Binding Sites
Polymerase Chain Reaction
Mutation
Temperature
Acids
Substrates

Keywords

  • Active site
  • Catalytic efficiency
  • Directed evolution
  • Kenaf
  • N-terminal region
  • Xylanase

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology

Cite this

Protein engineering of GH11 xylanase from Aspergillus fumigatus RT-1 for catalytic efficiency improvement on kenaf biomass hydrolysis. / Damis, Siti Intan Rosdianah; Abd. Murad, Abdul Munir; Diba Abu Bakar, Farah; Rashid, Siti Aishah; Jaafar, Nardiah Rizwana; Illias, Rosli Md.

In: Enzyme and Microbial Technology, Vol. 131, 109383, 01.12.2019.

Research output: Contribution to journalArticle

Damis, Siti Intan Rosdianah ; Abd. Murad, Abdul Munir ; Diba Abu Bakar, Farah ; Rashid, Siti Aishah ; Jaafar, Nardiah Rizwana ; Illias, Rosli Md. / Protein engineering of GH11 xylanase from Aspergillus fumigatus RT-1 for catalytic efficiency improvement on kenaf biomass hydrolysis. In: Enzyme and Microbial Technology. 2019 ; Vol. 131.
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AU - Diba Abu Bakar, Farah

AU - Rashid, Siti Aishah

AU - Jaafar, Nardiah Rizwana

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