【For INTERNATIONAL COLLABORATIVE RESEARCH 2022】

SUMMARY of

DATE: 4 May 2023

2022 RESEARCH RESULTS REPORTFor International Collaborative Research with IPR, Osaka University
Research Title

Crystallization and X-ray analysis of lignocellulose-degrading enzyme fromIndonesia local isolate

ApplicantName

Prof. Dr. Ni Nyoman Tri Puspaningsih, M.Si.

Affiliation(1) Department of Chemistry, Faculty of Science and Technology,
Universitas Airlangga, INDONESIA(2) University-CoE-Research Center for Bio-Molecule Engineering,
Universitas Airlangga, INDONESIA

Present Title

Professor

Research Collaborator (Host PI)

Prof. Genji KURISU

Summary
GbtXyl43B and CelGH5 are lignocellulose-degrading enzyme group that has a high novelty inenzyme structure. Understanding the structural interactions between enzyme and its substrate can beused as a basis for engineering the properties of enzymes to improve the ability and capability ofenzymes catalytic processes. Three-dimensional structure of enzyme can explain the catalytic
mechanism of enzyme against substrate, so that it can be explained how the enzyme interacts withthe substrate and the residues involved. Therefore, this study was conducted to determine thethree-dimensional structure of GbtXyl43B and CelGH5, and also its interaction with thesubstrate/ligand by using an X-ray crystallographic approach. The specific aims of this research are;(1) optimization of enzyme crystal formation and (2) exploring x-ray crystallographic data. The
method used in this study includes the stages of enzyme production, enzyme purification, andcrystallization screening & x-ray diffraction. Based on screening result using crystal screen kit,
GbtXyl43B protein produces significant aggregation, so that it does not produce a crystal profile likethe screening result of CelGH5. The results of CelGH5 screening provide information that it has thepotential result for further processed to obtain crystal diffraction data. While GbtXyl43B must be
re-optimized for getting the best conditions to produce crystals formed. The positive conditions ofthe formation of CelGH5 crystals were optimized to obtain a large crystal size and feasible foranalysis using X-ray diffraction. The results of X-ray diffraction at Spring 8 on the optimized
CelGH5 crystals formed, indicated that the CelGH5 crystals obtained were in a polycrystal state, soit is necessary for further optimization. The CelGH5 protein derived by the metagenome approachfrom compost soil of palm oil waste has enormous potential to be crystallized and to determine its
three-dimensional protein structure. So, research related to determining the structure of CelGH5
needs to be continued in order to obtain optimal results in this collaboration. This research is stillongoing in order to obtain proper results and can be published scientifically, so that the achievementof this research in the form of research papers/articles cannot be achieved. The next researcher whocontinues this research activity and is still a research team, is expected to be able to achieve thedesired results so that the expected joint scientific publications can be achieved.
*Deadline: May 12, 2023
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