Araştırma Çıktıları

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Author: search.filters.author.Yenenler, Asli

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    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.
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    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.
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    A possible founder mutation in FZD6 gene in a Turkish family with autosomal recessive nail dysplasia
    (BMC, 2019-01-01) Saygi, Ceren; Alanay, Yasemin; Sezerman, Ugur; Yenenler, Asli; Ozoren, Nesrin
    BackgroundAutosomal recessive nail dysplasia is characterized by thick and hard nails with a very slow growth on the hands and feet. Mutations in FZD6 gene were found to be associated with autosomal recessive nail dysplasia in 2011. Presently, only seven mutations have been reported in FZD6 gene
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    Design and characterizations of two novel cellulases through single-gene shuffling of Cel12A (EG3) gene from Trichoderma reseei
    (OXFORD UNIV PRESS, 2016-01-01) Yenenler, Asli; Sezerman, Osman Ugur
    Cellulases have great potential to be widely used for industrial applications. In general, naturally occurring cellulases are not optimized and limited to meet the industrial needs. These limitations lead to demand for novel cellulases with enhanced enzymatic properties. Here, we describe the enzymatic and structural properties of two novel enzymes, EG3\_S1 and EG3\_S2, obtained through the single-gene shuffling approach of Cel12A(EG3) gene from Trichoderma reseei. EG3\_S1 and EG3\_S2 shuffled enzymes display 59 and 75\% identity in protein sequence with respect to native, respectively. Toward 4-MUC, the minimum activity of EG3\_S1 was reported as 5.9-fold decrease in native at 35 degrees C, whereas the maximum activity of EG3\_S2 was reported as 15.4-fold increase in native activity at 40 degrees C. Also, the diminished enzyme activity of EG3\_S1 was reported within range of 0.6-to 0.8-fold of native and within range of 0.5-to 0.7-fold of native toward CMC and Na-CMC, respectively. For EG3\_S2 enzyme, the improved enzymatic activities within range of 1.1- to 1.4-fold of native and within range of 1.1-to 1.6-fold of nativewere reported toward CMC and Na-CMC, respectively. Moreover, we have reported 6.5-fold increase in the kcat/Km ratio of EG3\_S2 with respect to native and suggested EG3\_S2 enzyme as more efficient catalysis for hydrolysis reactions than its native counterpart.