Araştırma Çıktıları

Permanent URI for this communityhttps://hdl.handle.net/11443/931

Browse

Author: search.filters.author.Yenenler, AsliSubject: search.filters.subject.Trichoderma reesei

Search Results

Now showing 1 - 2 of 2
  • Thumbnail Image
    Item
    Enhancing Enzymatic Properties of Endoglucanase I Enzyme from Trichoderma Reesei via Swapping from Cellobiohydrolase I Enzyme
    (MDPI, 2019-01-01) Yenenler, Asli; Kurt, Hasan; Sezerman, Osman Ugur
    Utilizing plant-based materials as a biofuel source is an increasingly popular attempt to redesign the global energy cycle. This endeavour underlines the potential of cellulase enzymes for green energy production and requires the structural and functional engineering of natural enzymes to enhance their utilization. In this work, we aimed to engineer enzymatic and functional properties of Endoglucanase I (EGI) by swapping the Ala43-Gly83 region of Cellobiohydrolase I (CBHI) from Trichoderma reesei. Herein, we report the enhanced enzymatic activity and improved thermal stability of the engineered enzyme, called EGI\_swapped, compared to EGI. The difference in the enzymatic activity profile of EGI\_swapped and the EGI enzymes became more pronounced upon increasing metal-ion concentrations in the reaction media. Notably, the engineered enzyme retained a considerable level of enzymatic activity after thermal incubation for 90 min at 70 degrees C while EGI completely lost its enzymatic activity. Circular Dichroism spectroscopy studies revealed distinctive conformational and thermal susceptibility differences between EGI\_swapped and EGI enzymes, confirming the improved structural integrity of the swapped enzyme. This study highlights the importance of swapping the metal-ion coordination region in the engineering of EGI enzyme for enhanced structural and thermal stability.
  • Thumbnail Image
    Item
    Understanding the impacts of self-shuffling approach on structure and function of shuffled endoglucanase enzyme via MD simulations
    (WALTER DE GRUYTER GMBH, 2020-01-01) Yenenler, Asli; Gerlevik, Umut; Sezerman, Ugur
    Objective: We identify the impacts of structural differences on functionality of EG3\_S2 endoglucanase enzyme with MD studies. The results of previous experimental studies have been explained in details with computational approach. The objective of this study is to explain the functional differences between shuffled enzyme (EG3\_S2) and its native counterpart (EG3\_nat) from Trichoderma reseei, via Molecular Dynamics approach. Materials and methods: For this purpose, we performed MD simulations along 30 ns at three different reaction temperatures collected as NpT ensemble, and then monitored the backbone motion, flexibilities of residues, and intramolecular interactions of EG3\_S2 and EG3\_nat enzymes. Results: According to MD results, we conclude that EG3 S2 and EG3\_nat enzymes have unique RMSD patterns, e.g. RMSD pattern of EG3\_S2 is more dynamic than that of EG3\_nat at all temperatures. In addition to this dynamicity, EG3 S2 establishes more salt bridge interactions than EG3\_nat. Conclusion: By taking these results into an account with the preservation of catalytic Glu residues in a proper manner, we explain the structural basis of differences between shuffled and native enzyme via molecular dynamic studies.