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"Desain Struktur Mutan Α-Amilase Saccharomycopsis Fibuligera R64 Untuk Meningkatkan Adsorptivitasnya Terhadap Substrat Dengan Menggunakan Metode Bioinformatika"

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α-Amilase merupakan salah satu enzim penting yang digunakan pada industri
berbasis pati. Pada prosesnya, suhu tinggi dibutuhkan untuk memecah molekul
pati sehingga meningkatkan biaya produksi. Tingginya adsorptivitas α-amilase
terhadap substrat diketahui dapat menurunkan suhu hidrolisis. Modul pengikat
karbohidrat (CBM) dan/atau sisi pengikatan di permukaan (SBS) merupakan dua
bagian penting yang bertanggung jawab pada pengikatan substrat. α-Amilase
Saccharomycopsis fibuligera R64 (Sfamy R64), enzim asal Indonesia, diketahui
memiliki aktivitas amilolitik yang tinggi namun memiliki adsorptivitas yang
rendah terhadap substrat. Penelitian ini bertujuan untuk merancang mutan Sfamy
R64 agar memiliki adsorptivitas yang lebih baik dengan menambahkan SBS
pada strukturnya menggunakan metode bioinformatika. Perilaku struktural
Sfamy R64 dan kontrol positif (α-amilase Aspergillus niger) dipelajariv
menggunakan simulasi dinamika molekul (DM). Setelah itu, mutan Sfamy R64
dirancang untuk memiliki SBS yang stabil dengan meniru kontrol positif.
Afinitas substrat dievaluasi menggunakan metode MM/GBSA. Mutan Sfamy
R64 dengan konstruksi S383Y/S386W/N421G/S278N/A281K/Q384K/K398R
dan insersi G400_S401insTDGS stabil selama simulasi dan substrat dapat terikat
hingga 55 ns. Mutan dan kontrol positif memiliki perilaku SBS serupa dan energi
interaksi pada natif, mutan, dan kontrol positif masing-masing -5,2; -8,2; dan -
17,6 kkal/mol. Peningkatan pengikatan substrat yang terjadi pada mutan ini
berpotensi untuk diuji pada laboratorium.
Kata kunci: α-amilase, Sfamy R64, sisi pengikatan permukaan, adsorptivitas
pati, simulasi dinamika molekul

α-Amylase is one of the important enzymes in the starch-processing industry. In
that processing, high temperature is needed to break down the starch molecules
thus resulting in high cost of production. The high adsorptivity of α-amylase
toward substrate is known to decrease the hydrolysis temperature. Carbohydrate
Binding Module (CBM) and/or Surface Binding Site (SBS) are two important
part which responsible to the adsorption of starch. Saccharomycopsis fibuligera
R64 α-amylase (Sfamy R64), a locally-sourced enzyme from Indonesia, showed
high amylolytic activity but low starch adsorptivity. This study aimed to design
a mutant of Sfamy R64 to have better adsorptivity by introducting a SBS on its
structure using bioinformatics approach. The structural behavior of Sfamy R64
and positive control (Aspergillus niger α-amylase) were studied using molecular
dynamics simulation. After that, the mutants of Sfamy R64 were designed to
have a stable SBS which mimic the positive control. The substrate affinity was
evaluated using MM/GBSA method. Mutant of Sfamy R64 that constracted by
S383Y/S386W/N421G/S278N/A281K/Q384K/K398R and insertions of
G400_S401insTDGS was stable throughout DM simulation and substrate could
bound to the SBS over 55 ns. Mutant and positive control have similar structural
behavior of SBS and interaction energy of wild-type, mutant, and positive
control were -5.2, -8.2, and -17.6 kcal/mol, respectively. The enhanced substrate
binding in the mutant suggests the potential to be tested in laboratory.
Keywords: α-amylase, Sfamy R64, surface binding site, starch adsorptivity,
molecular dynamics simulation

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TM560

Publisher

Bandung : ., 2018

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Indonesia

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NONE

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