EKSTRAKSI DAN IDENTIFIKASI METABOLIT SEKUNDER DARI ISOLAT AL6 SERTA POTENSINYA SEBAGAI ANTIBAKTERI TERHADAP Escherichia coli

Alfian Syarifuddin, Sodiq Kamal, Fitriana Yuliastuti, Missya Putri Kurnia Pradani, Ni Made Ayu Nila Septianingrum

Abstract


Extraction and Identification of Secondary Metabolites from AL6 Isolates and Its Potential as Antibacterial against Escherichia coli

ABSTRACT

Secondary metabolites in the form of antibiotics can be produced by rhizospheric bacteria. AL6 bacterial isolate, which is one of the bacterial isolates from the rhosphere of Saccarum officinarum L., is known to produce antibiotic compounds. This study aims to determine the activity of antibiotics from AL6 ethyl acetate extracts produced by AL6 bacterial isolates, to analyze the minimum inhibitory concentration (MIC) and the similarity of the active substances using GCMS. The ethyl acetate extract obtained was tested for MIC at 1.25%, 2.5%, 5.0%, 10.0%, 20%, and 40% concentrations. Detection of potential antibiotic spots was carried out using bioautographic thin layer chromatography (TLC). Compounds responsible for antibiotic activity were analyzed using GCMS. Minimum inhibitory levels obtained reached 2.5%. The active spots responsible for antibiotic activity against Escherichia coli at Rf 0.94. Components detected using GCMS and suspected to be antibiotics include chloroform; ethane, 1,1-dimethoxy-(CAS) dimethyl acetal; dan 1,3-dioxolane, 2-methoxymethyl-2,4,5-trimethyl.

Keywords: AL6 bacterial isolate; antibiotic; Escherichia coli; GCMS; MIC

ABSTRAK

Metabolit sekunder berupa antibiotik dapat diproduksi oleh bakteri rizosfer. Isolat bakteri AL6, salah satu isolat bakteri dari rizosfer Saccarum officinarum L., diketahui dapat menghasilkan senyawa antibiotik. Penelitian ini bertujuan mengetahui aktivitas antibiotik dari ekstrak etil asetat antibiotik AL6 yang dihasilkan isolat bakteri AL6, menganalisis kadar hambat minimum (KHM), serta kemiripan zat aktif menggunakan GCMS. Ekstrak etil asetat yang diperoleh diuji KHM-nya pada konsentrasi 1,25%, 2,5%, 5,0%, 10,0%, 20%, dan 40%. Deteksi bercak yang berpotensi sebagai antibiotik dilakukan menggunakan kromatografi lapis tipis (KLT) bioautografi. Senyawa yang berperan dalam aktivitas antibiotik dianalisis menggunakan GCMS. Kadar hambat minimal yang diperoleh mencapai 2,5%. Hasil uji KLT bioautografi memperlihatkan bercak aktif sebagai antibiotik terhadap Escherichia coli pada Rf 0,94. Komponen senyawa yang terdeteksi menggunakan GCMS dan diduga sebagai antibiotik antara lain chloroform; ethane, 1,1-dimethoxy-(CAS) dimethyl acetal; dan 1,3-dioxolane, 2-methoxymethyl-2,4,5-trimethyl.


Keywords


antibiotik; Escherichia coli; GCMS; isolat bakteri AL6; KHM

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References


Alimuddin, Widada J, Asmara W, Mustofa (2011) Antifungal production of a strain of Actinomycetes spp. isolated from the rhizosphere of cajuput plant: Selection and detection of exhibiting activity against tested fungi. Indones J Biotechnol 16:1-10. doi: 10.22146/ijbiotech.7829

Ahsan T, Chen J, Wu Y, Irfan M, Shafi J (2017) Screening, identification, optimization of fermentation conditions, and extraction of secondary metabolites for the biocontrol of Rhizoctonia solani AG-3. Biotechnol Biotechnol Equip 31:91–98. doi: 10.1080/13102818.2016.1259016

Apsari PP, Budiarti S, Wahyudi AT (2019) Actinomycetes of rhizosphere soil producing antibacterial compounds against urinary tract infection bacteria. Biodiversitas J Biol Divers 20:1259–1265. doi: 10.13057/biodiv/d200504

Chasanah E, Noor NM, Risjani Y, Dewi AS (2012) Aktivitas Antibakteri Dan Antioksidan Ekstrak Streptomyces sp. dan Exserohilum rostratum yang Dikultivasi Pada Tiga Jenis Medium Pertumbuhan. J Pascapanen Bioteknol Kelaut Perikan 7:39. doi: 10.15578/jpbkp.v7i1.67

Fauziyya R, Nurani LH, Sulistyani N (2017) antibacterial compound identification of cayenne pepper leaf extract (Capsicum frutescens L.) against Klebsiella pneumoniae and cell leakage mechanism. Maj Obat Tradis 22:166. doi: 10.22146/mot.31550

Hassan AS (2014) The antibacterial activity of dimethyl sulfoxide (DMSO) with and without of some ligand complexes of the transitional metal ions of ethyl coumarin against bacteria isolate from burn and wound infection. J Nat Sci Res 4:106. doi: 10.13140/RG.2.2.36692.40321Gebreyohannes G, Moges F, Sahile S, Raja N (2013) Isolation and characterization of potential antibiotic producing actinomycetes from water and sediments of Lake Tana, Ethiopia. Asian Pac J Trop Biomed 3:426–435. doi: 10.1016/S2221-1691(13)60092-1

Hemashenpagam N (2011) Purification of secondary metabolites from soil actinomycetes. Int J Microbiol Res 3:148–156. doi: 10.9735/0975-5276.3.3.148-156

Khucharoenphaisan K, Sripairoj N, Sinma K (2012) Isolation and identification of actinomycetes from termite’s gut against human pathogen. Asian J Anim Vet Adv 7:68–73. doi: 10.3923/ajava.2012.68.73

Mohamed H, Miloud B, Zohra F, García-Arenzana JM, Veloso A, Rodríguez-Couto S (2017) Isolation and characterization of actinobacteria from Algerian sahara soils with antimicrobial activities. Int J Mol Cell Med 6:109–120

Mulyadi, Sulistyani N (2013) Aktivitas cairan kultur 12 isolat actinomycetes terhadap bakteri resisten. J Kesehat Masy J Public Health 7:89–96. doi: 10.12928/kesmas.v7i2.1043

Nanjwade B, Chandrashekhara S, Goudanavar P, Shamarez A, Manvi F (2010) Production of antibiotics from soil-isolated actinomycetes and evaluation of their antimicrobial activities. Trop J Pharm Res 9:373–377. doi: 10.4314/tjpr.v9i4.58933

Narwanti I, Sulistyani N (2015) TLC-bioautography profile of ethyl acetate extract of 5 bacteria isolated from Ficus carica L rhizosphere. Int J Public Health Sci IJPHS 4:81-87. doi: 10.11591/.v4i2.4716

Raghava Rao KV, Mani P, Satyanarayana B, Raghava Rao T (2017) Purification and structural elucidation of three bioactive compounds isolated from Streptomyces coelicoflavus BC 01 and their biological activity. 3 Biotech 7:1–12. doi: 10.1007/s13205-016-0581-9

Rai M, Bhattarai N, Dhungel N, Mandal PK (2016) Isolation of antibiotic producing Actinomycetes from soil of Kathmandu valley and assessment of their antimicrobial activities. Int J Microbiol Allied Sci 2:22–26

Retnowati Y, Moeljopawiro S, Djohan TS, Soetarto ES (2018) Antimicrobial activities of actinomycete isolates from rhizospheric soils in different mangrove forests of Torosiaje, Gorontalo, Indonesia. Biodiversitas 19:2196–2203. doi: 10.13057/biodiv/d190627

Salim FM, Sharmili SA, Anbumalarmathi J, Umamaheswari K (2017) Isolation, molecular characterization and identification of antibiotic producing actinomycetes from soil samples. J Appl Pharm Sci 7:069–075. doi: 10.7324/JAPS.2017.70909

Selvameenal L, Radhakrishnan M, Balagurunathan R (2009) Antibiotic pigment from desert soil actinomycetes; biological activity, purification and chemical screening. Indian J Pharm Sci 71:499–504. doi: 10.4103/0250-474X.58174

Singh LS, Sharma H, Talukdar NC (2014) Production of potent antimicrobial agent by actinomycete, Streptomyces sannanensis strain SU118 isolated from phoomdi in Loktak Lake of Manipur, India. BMC Microbiol 14:1-13. doi: 10.1186/s12866-014-0278-3

Sulistyani N, Akbar AN (2014) Aktivitas isolat actinomycetes dari rumput laut (Eucheuma cottonii) sebagai penghasil antibiotik terhadap Staphylococcus aureus dan Escherichia coli. J Ilmu Kefarmasian Indones 12:1-9

Syarifuddin A, Sulistyani N (2018) Activity of antibiotic bacterial isolate kp13 and cell leakage analysis of Escherichia coli bacteria. J Ilmu Kefarmasian Indones 16:137–144. doi: 10.35814/jifi.v16i2.529

Syarifuddin A, Sulistyani N (2019) Karakterisasi fraksi teraktif senyawa antibiotik isolat kp 13 dengan metode densitometri dan klt-semprot. J Ilm Ibnu Sina 4:156–166. doi: 10.36387/jiis.v4i1.263

Syarifuddin A, Sulistyani N, Kintoko (2019) Profil KLT-bioautografi dan densitometri fraksi teraktif (isolat kp13) dari bakteri rizosfer kayu putih (Melaleucaleucadendron L.). J Farm Sains Prakt 5:21–25. doi: 10.31603/pharmacy.v5i1.2291

Utomo SB, Fujiyanti M, Lestari WP, Mulyani S (2018) Antibacterial activity test of the C-4-methoxy phenylcalix[4]resorcinarene compound modified by hexadecyltrimethylammonium-bromide against Staphylococcus aureus and Escherichia coli bacteria. J Kim Pendidik Kim 3:201. doi: 10.20961/jkpk.v3i3.22742

Wang X, Huang L, Kang Z, Buchenauer H, Gao X (2010) Optimization of the fermentation process of actinomycete strain Hhs.015T. J Biomed Biotechnol 2010:1–10. doi: 10.1155/2010/141876




DOI: http://dx.doi.org/10.29122/jbbi.v6i2.3516

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