Karakterisasi Bioplastik Komposit Dari Tepung Biji Durian Dan Tepung Porang Dengan Penambahan Ekstrak Etanol Daun Pucuk Merah (S. Myrtifolium) Sebagai Bahan Antibakteri Dan Uji In Silico

Permatasari, Nelsy Dian and M.App.Sc, Dr. Ir. Sudarminto Setyo Yuwono and S.Si,M.Si,Ph.D, Masruri (2022) Karakterisasi Bioplastik Komposit Dari Tepung Biji Durian Dan Tepung Porang Dengan Penambahan Ekstrak Etanol Daun Pucuk Merah (S. Myrtifolium) Sebagai Bahan Antibakteri Dan Uji In Silico. Doktor thesis, Universitas Brawijaya.

Abstract

Plastik sintetis telah menimbulkan permasalahan dan tantangan yang signifikan bagi dunia saat ini karena membutuhkan waktu lama untuk terurai, tidak ramah lingkungan dan mencemari manusia secara tidak langsung. Diperlukan plastik ramah lingkungan yang terbuat dari bahan alam. Umumnya, plastik ramah lingkungan terbuat dari bahan terbarukan seperti pati atau tepung utuh. Salah satu bahan alam yang digunakan yaitu biji durian yang dapat diubah menjadi tepung biji durian karena tepung durian mengandung pati, karbohidrat, protein, lemak dan serat yang dapat digunakan sebagai bahan bioplastik. Pembuatan bioplastik dari tepung saja sudah pernah dilakukan namun menghasilkan karakteristik fisik yang rendah sehingga diperlukan bahan lain. Bahan lain yang digunakan adalah tepung porang. Tepung porang dapat berperan sebagai binder atau pengikat, thickening, gelling, texturing dan sebagai pembentuk film. Upaya peningkatan sifat antibakteri dari bioplastik ditambahkan ekstrak etanol daun S.myrtifolium pada bioplastik. S.myrtifolium mengandung senyawa antibakteri yang dapat menghambat pertumbuhan bakteri E.coli secara in vitro. Penambahan ekstrak etanol daun S.myrtifolium pada bioplastik belum pernah dilakukan sehingga merupakan keterbaruan pada penelitian ini. Pembuktian aktivitas secara komputasi/ virtual belum ada yang melakukan sehingga diperlukan pengujian secara in silico untuk mengetahui mekanisme penghambatan ekstrak etanol daun S. myrtifolium sebagai bahan bioaktif dalam menghambat pertumbuhan bakteri E. coli. Penelitian Tahap 1 bertujuan : mengetahui, menganalisis dan mengevaluasi karakteristik fisik, kimia, fungsional dan struktural tepung biji durian var.tembaga dan tepung biji durian komersial. Karakteristik ini meliputi proksimat, pati, amilosa, amilopektin, ukuran partikel, gugus fungsi, pola kristalinitas, mikrostruktur, sifat termal, sifat pasting. Penelitian Tahap 2 bertujuan : mengetahui, menganalisis dan mengevaluasi pengaruh penambahan tepung biji durian, tepung porang dan ekstrak etanol daun pucuk merah terhadap sifat fisik bioplastik dan sifat antibakteri bioplastik menggunakan metode Faktorial Design menggunakan Design Expert DX.10.0.0 dengan 3 faktor yaitu tepung biji durian (X1), tepung porang (X2) dan dan ekstrak etanol daun pucuk merah (X3) menghasilkan 8 percobaan setelah dioperasikan dengan program Design Expert DX 10.0.0. batas bawah (-1) dan batas atas (+1) untuk tepung biji durian adalah 0,5 dan 1 % w/v. Batas bawah (-1) dan batas atas (+1) untuk tepung porang adalah 0,5 dan 1 % w/v dan batas bawah (-1) dan batas atas (+1) dari ekstrak etanol daun pucuk merah adalah 5 dan 25 %v/v. Respon (Y) yang diukur adalah kekuatan tarik (Y1), perpanjangan (Y2) dan zona hambat (Y3). Pengujian aktivitas antibakteri dilakukan dengan uji zona hambat menggunakan bakteri E.coli. suspensi bakteri disebar dalam plate agar, film dipotong dan diletakkan pada plate agar. Plate diinkubasi selama 12 jam pada suhu 37°C. Diameter zona hambat diukur dan diulang sebanyak tiga kali. Kontrol yaitu bioplastik dengan penambahan natrium benzoat, perlakuan terbaik adalah bioplastik antibakteri terbaik kemudian dibandingkan dengan bioplastik yang terbuat dari tepung porang (TP) dan bioplastik dari bahan tepung biji durian (TBD) dan tepung porang (TP). Tahap 3 bertujuan : mengetahui, menganalisis dan mengevaluasi mekanisme penghambatan senyawa antibakteri dari ekstrak etanol daun pucuk merah dengan metode in silico untuk memprediksi aktivitas dan mekanisme senyawa antibakteri pada daun pucuk merah secara virtual dengan menggunakan data primer (berdasarkan hasil pengujian LCMS). Berdasarkan pengujian LCMS diperoleh empat senyawa yaitu 2-Propanon,1,3-bis(5-nitro-2-furanil), Quercetin-3-O-alpha-L-arabinopyranoside, Calopiptin dan Auraptenol. Menentukan protein target yang akan digunakan. Protein target yang digunakan yaitu Alr (alanine racemase) dan TG yang diunduh dari database Protein Data Bank (PDB). Protein alanine racemase (4WR3) dan transglikosilase (1SLY) sebagai reseptor diunduh dari database PDB. Alanin rasemase (Alr) dan transglikosilase (TG) diprediksi ikatannya dengan menggunakan Molegro virtual docker versi 5. Struktur 3D ligan 2-Propanon, 1,3-bis(5-nitro-2-furanil) (CID152537), Quercetin-3-O-alpha-L-arabinopyranoside (CID 5481224), Calopiptin (CID 12302265) dan Auraptenol (CID 13343540) diunduh dari database PubChem. Quercetin-3-O-beta-D-glucoronide secara komputasi dimodelkan oleh server web Cheminfo (http://www.cheminfo.org/) dan MolView (https://molview.org/). Reseptor diikatkan ke ligan menggunakan Molegro Virtual Docker 5 dengan luas spesifik alanin racemase sebagai berikut, X=43,87; Y=2,45; Z=2,99; Radius 9 dan transglikosilase X=10.52 Y=47.19; Z=57,06 Radius 13. Parameter untuk docking adalah MolGrid 0.3A, pengulangan docking 10x, dan RMSD < 2. Memvalidasi hasil docking daun Pucuk Merah, dilakukan docking reseptor dengan kontrol antibakteri Sodium Benzoate (CID243) dari PubChem, Pyridoxal-5'-Phosphate (PDB ID 4WR3) untuk alanine racemase dan 4-O-(4-O-Sulfonyl- N-Acetylglucosamininyl)-5-Methylhydroxy-L-Proline-Taurine (PDB ID 1SLY) untuk transglikosilase. Hasil docking dianalisis dengan PyMol 2.2 dan Discovery Studio 21 Hasil penelitian tahap I diperoleh tepung biji durian var.tembaga (TBDT) berbeda nyata dengan tepung biji durian komersial (TBDK) pada parameter protein, lemak, kadar karbohidrat, serat kasar, amilosa, swelling power, OAC, WAC, L,a,b derajat putih. TBDT dan TBDK tidak berbeda nyata pada parameter kadar abu, kadar air, kadar pati dan kelarutan. Secara mikrostruktur TBDT memiliki permukaan kasar yang dilapisi oleh lapisan tipis. Sedangkan pada TBDK tidak dilapisi lapisan tipis. TBDT memiliki ukuran partikel lebih kecil dan memiliki suhu gelatinoisasi lebih tinggi daripada TBDK. TBDT dan TBDK memiliki pola spektra yang sama berdasarkan uji FTIR. TBDT dan TBDK memiliki pola kristalinitas yang sama yaitu pola difraksi tipe A. TBDK memiliki nilai viskositas breakdown, viskositas setback, waktu puncak dan suhu pasting lebih tinggi dibandingkan dengan TBDT. Namun memiliki viskositas trough dan viskositas final lebih rendah berdasarkan uji pasting berdasarkan RVA. Hasil penelitian tahap II diperoleh perlakuan terbaik didapatkan pada massa tepung biji durian 0,5% b/v , massa tepung porang 1% b/v dan konsentrasi ekstrak etanol daun S.myrtifolium 25% v/v menghasilkan kuat tarik 3,30 MPa, elongasi 50% dan zona hambat 15,33 mm. Selain itu pada kombinasi itu mempunyai ketebalan 0,115 mm, modulus young 0,066 MPa, densitas 1,37 g/cm3 kadar air 17,14% dan kelarutan 76,91 %. Hasil penelitian tahap III diperoleh senyawa bioaktif khususnya flavonoid dalam S. myrtifolium berpotensi sebagai antibakteri. Mekanisme antibakteri flavonoid S. myrtifolium adalah melalui penghambatan biosintesis dinding sel.

English Abstract

Synthetic plastics have caused significant problems and challenges for the world today because they take a long time to decompose, are not environmentally friendly and pollute humans indirectly. Environmentally friendly plastic made from natural materials is required. Generally, eco-friendly plastics are made from renewable materials such as starch or whole flour. One of the natural ingredients used is durian seeds which can be converted into durian seed flour because durian seed flour contains starch, carbohydrates, protein, fat and fiber which can be used as bioplastic materials. The manufacture of bioplastics from flour alone has been done but produces low physical characteristics so that other materials are needed. Another ingredient used is porang flour. Porang flour can act as a binder, thickening, gelling, texturing and as a film former. Efforts to improve the antibacterial properties of bioplastics added ethanol extract of S.myrtifolium leaves to bioplastics. S.myrtifolium contains antibacterial compounds that can inhibit the growth of E.coli bacteria in vitro. The addition of ethanolic extract of S.myrtifolium leaves to bioplastics has never been done so it is a novelty in this study. There is no evidence of computational/virtual activity, so in silico testing is needed to determine the mechanism of inhibition of the ethanolic extract of S. myrtifolium leaves as a bioactive ingredient in inhibiting the growth of E. coli bacteria. The stage I of research aims: to determine, analyze and evaluate the physical, chemical, functional and structural characteristics of durian seed flour var.tembaga (TDSF) and commercial durian seed flour (CDSF). These characteristics include proximate, starch, amylose, amylopectin, particle size, functional group, crystallinity pattern, microstructure, thermal properties and pasting properties. The stage II of research aims: to determine, analyze and evaluate the effect of adding durian seed flour, porang flour and ethanolic extract of S.myrtifolium leaves to the physical properties of bioplastics and antibacterial properties of bioplastics using the Factorial Design method using Design Expert DX.10.0.0 with 3 factors, namely durian seed flour var.tembaga (X1), porang flour (X2) and ethanolic extract of S.myrtifolium leaves (X3) resulted in 8 trials after being operated with the Design Expert DX 10.0.0 program. The lower limit (-1) and upper limit (+1) for durian seed flour were 0.5 and 1% w/v, respectively. The lower limit (-1) and upper limit (+1) for porang flour were 0.5 and 1% w/v and the lower limit (-1) and upper limit (+1) of the ethanolic extract of S.myrtifolium were 5 and 25 %v/v. The responses (Y) that were measured were tensile strength (Y1), elongation (Y2) and zone of inhibition (Y3). Testing of antibacterial activity was carried out by testing the zone of inhibition using E.coli bacteria. The bacterial suspension was spread on the agar plate, the film was cut and placed on the agar plate. Plates were incubated for 12 hours at 37°C. The diameter of the inhibition zone was measured and repeated three times. The control is bioplastic with the addition of sodium benzoate, the best treatment is the best antibacterial bioplastic then compared with bioplastics made from porang flour (PF) and bioplastic from a mixture of durian seed flour (DSF) and porang flour (PF). The stage III aims: to determine, analyze and evaluate the mechanism of inhibition of antibacterial compounds from the ethanolic extract of S.myrtifolium with the in silico method to predict the activity and mechanism of antibacterial compounds in S.myrtifolium virtually using primary data (based on the results of the LCMS test). Based on the LCMS test, four compounds were obtained, namely 2-Propanone, 1,3-bis(5-nitro-2-furanyl), Quercetin-3-O-alpha-L-arabinopyranoside, calopiptin and auraptenol. Determine the target protein to be used. The target proteins used were Alr (alanine racemase) and TG which were downloaded from the Protein Data Bank (PDB) database. Protein alanine racemase (4WR3) and transglycosylase (1SLY) as receptors were downloaded from the PDB database. Alanine racemase (Alr) and transglycosylase (TG) were predicted for their binding using Molegro virtual docker version 5. 3D structure of the ligand 2-Propanone, 1,3-bis(5-nitro-2-furanyl) (CID152537), Quercetin-3-O -alpha-L-arabinopyranoside (CID 5481224), Calopiptin (CID 12302265), and Auraptenol (CID 13343540) were downloaded from the PubChem database. Quercetin-3-O-beta-D-glucoronide is computationally modeled by the Cheminfo web servers (http://www.cheminfo.org/) and MolView (https://molview.org/). The receptor was bound to the ligand using Molegro Virtual Docker 5 with a specific area of alanine racemase as follows, X=43.87; Y=2.45; Z=2.99; Radius 9 and transglycosylase X=10.52 Y=47.19; Z=57.06 Radius 13. Parameters for docking were MolGrid 0.3A, 10x docking repetition, and RMSD < 2. Validating the results of docking of Pucuk Merah leaves, docking the receptor with antibacterial control Sodium Benzoate (CID243) from PubChem, Pyridoxal-5' -Phosphate (PDB ID 4WR3) for alanine racemase and 4-O-(4-O-Sulfonyl-N-Acetylglucosamininyl)-5-Methylhydroxy-L-Proline-Taurine (PDB ID 1SLY) for transglycosylase. Docking results were analyzed with PyMol 2.2 and Discovery Studio 21. The results of the stage I of the research showed that the durian seed flour var.tembaga (TDSF) was significantly different from the commercial durian seed flour (CDSF) on the parameters of protein, fat, carbohydrate content, crude fiber, amylose, swelling power, OAC, WAC, L, a, b and degrees of whiteness. TDSF and CDSF were not significantly different in the parameters of ash content, moisture content, starch content and solubility. Microstructurally TDSF has a rough surface covered by a thin layer. Meanwhile, CDSF is not coated with a thin layer. TDSF has a smaller particle size and has a higher gelatinization temperature than CDSF. TDSF and CDSF have the same spectral pattern based on the FTIR test. TDSF and CDSF have the same crystallinity pattern, namely type A diffraction pattern. CDSF has higher breakdown viscosity, setback viscosity, peak time and pasting temperature than TDSF. However, it has lower trough viscosity and final viscosity based on pasting test based on RVA. The results of the stage II of the research showed that the best treatment was obtained at a mass of 0.5% w/v of durian seed flour, 1% w/v of porang flour and a concentration of 25% v/v S.myrtifolium leaf extract, which resulted in a tensile strength of 3.30 MPa, elongation of 50% and an inhibition zone of 15.33. mm. In addition, the combination has a thickness of 0.115 mm, Young's modulus of 0.066 MPa, a density of 1.37 g/cm3, a water content of 17.14% and a solubility of 76.91%. The results of the stage III of the study showed that bioactive compounds, especially flavonoids in S. myrtifolium, had potential as antibacterial. The antibacterial mechanism of S. myrtifolium flavonoids is through inhibition of cell wall biosynthesis.

Item Type:	Thesis (Doktor)
Identification Number:	0622100003
Uncontrolled Keywords:	Bioplastik, tepung biji durian, tepung porang, S.myrtifolium, antibakteri, in silico, bioplastic, durian seed flour, porang flour, S.myrtifolium, antibacterial, in silico
Subjects:	300 Social sciences > 338 Production > 338.1 Agriculture > 338.16 Production efficiency
Divisions:	S2/S3 > Doktor Teknologi Industri Pertanian, Fakultas Teknologi Pertanian
Depositing User:	soegeng Moelyono
Date Deposited:	19 May 2023 06:14
Last Modified:	19 May 2023 06:14
URI:	http://repository.ub.ac.id/id/eprint/199894

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