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Jurnal Bioteknologi & Biosains Indonesia (JBBI)
Published by Badan Pengkajian dan Penerapan Teknologi
ISSN : 24422606     EISSN : 2548611X     DOI : -
Core Subject : Science, Agriculture,
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Immunology & microbiology
JBBI, Indonesian Journal of Biotechnology & Bioscience, is published twice annually and provide scientific publication medium for researchers, engineers, practitioners, academicians, and observers in the field related to biotechnology and bioscience. This journal accepts original papers, review articles, case studies, and short communications. The articles published are peer-reviewed by no less than two referees and cover various biotechnology subjects related to the field of agriculture, industry, health, environment, as well as life sciences in general. Initiated at the then Biotech Centre, the journal is published by the Laboratory for Biotechnology, the Agency for the Assessment and Application of Technology, BPPT.
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EMBRIOGENESIS SOMATIK IN VITRO DAN REGENERASI PLANLET DARI TIGA VARIETAS ALFALFA (Medicago sativa L.) Sulastri, Sulastri; Nawfetrias, Winda; Pinardi, Djatmiko; Rosdayanti, Henti
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 6 No. 1 (2019): June 2019
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (142.917 KB) | DOI: 10.29122/jbbi.v6i1.3348

Abstract

In Vitro Somatic Embryogenesis and Plantlet Regeneration of Three Varieties of Alfalfa (Medicago sativa L.)ABSTRACTAlfalfa (Medicago sativa L.) is a valuable plant as a source of food for animal, forage, pharmaceutical, medicine, food supplement, and human consumption.  In vitro selection technology combined with induction or spontaneous mutagenesis has been effective in altering or isolating genetic variability for desirable characters.  Consequently, a reproducible in vitro propagation technique of that plant is mandatory. The aim of the research was to obtain information on the embryogenic callus induction, somatic embryogenesis, and plantlet regeneration of three varieties of alfalfa. The results showed that an optimum embryogenic callus induction (82%) was obtained on Murashige & Skoog (MS) basal medium containing 2 ppm 2,4-dichlorophenoxyacetic acid (2,4-D), 2 ppm kinetin and 2 ppm a-naphthaleneacetic acid (NAA). Those embryogenic calli could subsequently develop into somatic embryos, which germinated and regenerated into normal plantlets on R1 medium consisting of MS nutrients without the addition of plant growth regulator.Keywords: alfalfa, callus, embryogenic, plantlets, regeneration ABSTRAKAlfalfa (Medicago sativa) adalah tanaman berharga sebagai sumber makanan untuk hewan, yaitu hijauan pakan ternak, farmasi, obat-obatan, suplemen makanan dan konsumsi manusia. Teknologi seleksi in vitro yang dikombinasikan dengan induksi atau mutagenesis spontan telah terbukti efektif dalam mengubah atau mengisolasi variabilitas genetik untuk karakter yang diinginkan. Oleh sebab itu, keberhasilan teknik perbanyakan in vitro yang telah terbukti dapat direproduksi dari tanaman tersebut menjadi syarat yang harus terpenuhi. Tujuan dari penelitian ini adalah untuk mendapatkan informasi mengenai induksi kalus embriogenik, embriogenesis somatik dan regenerasi planlet dari tiga varietas alfalfa. Hasil penelitian menunjukkan bahwa induksi kalus embriogenik optimal (82%) didapat pada media Murashige & Skoog (MS) dengan  2 ppm 2,4-dichlorophenoxyacetic acid (2,4-D), 2 ppm kinetin dan 2 ppm a-naphthaleneacetic acid (NAA). Kalus embriogenik tersebut dapat membentuk embrio somatik, embrionya berkecambah dan beregenerasi membentuk planlet normal pada perlakuan media R1 yaitu nutrisi MS tanpa penambahan zat pengatur tumbuh.Kata Kunci: alfalfa, embriogenik, kalus, planlet, regenerasi
PENINGKATAN KUALITAS BIJI KAKAO (Theobroma cacao L) MELALUI FERMENTASI MENGGUNAKAN Lactobacillus sp. dan Pichia kudriavzevii Meryandini, Anja; Basri, Asrianti; Sunarti, Titi Candra
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 6 No. 1 (2019): June 2019
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (848.014 KB) | DOI: 10.29122/jbbi.v6i1.3048

Abstract

The Improvement of Cacao Beans Quality through Fermentation by Using Lactobacillus sp. and Pichia kudriavzeviiABSTRACTIndonesia is one of the main cacao producers in the world. Indonesian cacao product is, however, relatively of low quality. Quality improvement of cacao beans is thus needed to increase added value of the product through such method as fermentation using bacteria and yeast. This study was conducted using four fermentation treatments, namely F1 (spontaneous fermentation without the addition of inoculum), F2 (addition of lactic acid bacteria inoculum), F3 (addition of yeast inoculum), F4 (addition of mixed lactic acid bacteria and yeast inoculum). The fermentation was carried out for 5 days. The parameters measured were the microbial cell number, pH, ethanol, total reducing sugar, and total acid concentration, as well as cacao seed quality. Results showed that, compared to the other treatments, the F4 treatment gave the best result, namely 83% of the cacao seeds being fermented, 2% non-fermented, 14% unfermented, 1% moldy, and 2% germinated. The liquid produced during the fermentation contained the highest reducing sugar of 123.38 mg·mL-1, the highest total acid of 24.42 mg·mL-1, and 3.57% ethanol.Keywords: cacao beans, fermentation, lactic acid bacteria, starter, yeast ABSTRAKIndonesia adalah salah satu penghasil kakao utama di dunia. Namun berdasarkan mutu, produk kakao Indonesia masih relatif tergolong rendah. Peningkatan kualitas biji kakao diperlukan untuk memberikan nilai tambah pada produk melalui metode seperti fermentasi menggunakan bakteri dan khamir. Penelitian ini dilakukan dengan empat perlakuan fermentasi yaitu F1 (fermentasi secara spontan tanpa penambahan inokulum), F2 (dengan penambahan inokulum bakteri asam laktat (BAL)), F3 (dengan penambahan inokulum khamir), F4 (dengan penambahan inokulum campuran bakteri asam laktat dan khamir). Fermentasi dilakukan selama 5 hari, dan parameter yang diukur selama fermentasi adalah jumlah mikroba, pH, kadar etanol, gula pereduksi, total asam serta kualitas biji. Hasil menunjukkan bahwa, dibandingkan perlakuan lainnya, perlakuan F4 memberikan hasil terbaik yaitu 83% biji terfermentasi, 2% tidak terfermentasi, 14% terfermentasi sebagian, 1% berjamur, dan 2% berkecambah. Cairan fermentasi tersebut mengandung gula reduksi yang paling tinggi 123,38 mg·mL-1, total asam tertinggi 24,42 mg·mL-1, dan kadar etanol mencapai 3,57%.Kata Kunci: bakteri asam laktat (BAL), biji kakao, fermentasi, khamir, starter
Back Cover JBBI Vol 6, No 1, June 2019 Sriherwanto, Catur
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 6 No. 1 (2019): June 2019
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (83.157 KB) | DOI: 10.29122/jbbi.v6i1.3618

Abstract

MIKROPROPAGASI TANAMAN TALAS BENENG (Xanthosoma undipes K. Koch) DENGAN PERLAKUAN BENZIL AMINOPURIN, TIAMIN, DAN ADENIN Sari, Laela; Wulansari, Aida; Noorrohmah, Siti; Ermayanti, Tri Muji
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 6 No. 1 (2019): June 2019
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29122/jbbi.v6i1.3216

Abstract

Micropropagation of Beneng Taro (Xanthosoma undipes K. Koch) with Benzyl Amino Purine, Thiamine, and Adenine TreatmentABSTRACTConventional production of Beneng taro seeds (Xanthosoma undipes K. Koch) is constrained by the limited number of tubers, thus an alternative solution is needed such as in vitro propagation. This study was aimed to obtain a micropropagation technique of Beneng taro on MS media with BAP, thiamine, and adenine treatment, and to determine its growth at the acclimatization stage. This research consisted of shoot multiplication and acclimatization. Shoot propagation was carried out on MS media with 8 treatments, namely ½MS and MS without addition of growth promoting substance, and MS with 1, 2 and 3 mg×L-1 BAP, with or without addition of 1 mg×L-1 thiamine and 2 mg×L-1 adenine. Each treatment was replicated four times, each consisting of four shoots. Growth observation was made from 1st to 5th week on petiole length, and number of shoots, leaves and roots. Acclimatization was carried out on soil media, compost, and husks in a ratio of 1: 1: 1. The results showed that the best media for shoot multiplication was MS + 1 mg×L-1 BAP + 1 mg×L-1 thiamine + 2 mg×L-1 adenine with an average of 3.5 shoots, while the best medium for the petiole length was ½MS with an average value of 6.97 cm. The results of acclimatization showed that 100% planlets survived, and plantlets grown on MS media + 3 mg×L-1 BAP had the highest number of shoots with an average of 4.2.Keywords: adenine, Beneng taro, benzil amino purine (BAP), micropropagation, thiamineABSTRAKPenyediaan bibit talas Beneng (Xanthosoma undipes K. Koch) secara konvensional terkendala terbatasnya jumlah umbi, sehingga perlu solusi alternatif, diantaranya melalui perbanyakan in vitro. Tujuan penelitian ini adalah untuk mendapatkan teknik mikropropagasi talas beneng pada media MS dengan perlakuan BAP, tiamin, adenin, dan untuk mengetahui pertumbuhannya pada tahap aklimatisasi. Penelitian ini meliputi perbanyakan tunas dan aklimatisasi. Perbanyakan tunas menggunakan media MS dengan 8 perlakuan yaitu ½MS dan MS tanpa penambahan zat pengatur tumbuh (ZPT), serta MS dengan 1, 2 dan 3 mg×L-1 BAP dengan atau tanpa penambahkan 1 mg×L-1 tiamin dan 2 mg×L-1 adenin. Setiap perlakuan mempunyai empat ulangan, setiap ulangan terdiri atas empat tunas. Pertumbuhan diamati mulai minggu ke-1 hingga ke-5 terhadap panjang petiol serta jumlah anakan, daun dan akar. Aklimatisasi dilakukan pada media tanah, kompos dan sekam dengan perbandingan 1:1:1. Hasil menunjukkan bahwa media terbaik perbanyakan tunas adalah MS + 1 mg×L-1 BAP + 1 mg×L-1 tiamin + 2 mg×L-1 adenin dengan rata-rata 3,5 tunas, sedangkan media terbaik untuk panjang tangkai daun adalah ½MS dengan nilai rata-rata 6,97 cm. Hasil aklimatisasi menunjukkan bahwa 100% planlet hidup dan planlet yang ditumbuhkan pada media MS + 3 mg×L-1 BAP mempunyai jumlah anakan terbanyak dengan rata-rata 4,2.Kata Kunci: adenine, benzil amino purin (BAP), mikropropagasi, talas Beneng, tiamine
TRANSFORMASI GENETIK DAN EKSPRESI MUTAN SUCROSE PHOSPHATE SYNTHASE PADA TANAMAN TOMAT Fibriani, Suwinda; Agustien, Inyana Dwi; Sawitri, Widhi Dyah; Sugiharto, Bambang
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 6 No. 1 (2019): June 2019
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (699.256 KB) | DOI: 10.29122/jbbi.v6i1.3341

Abstract

Genetic Transformation and Expression of Sucrose Phosphate Synthase Mutant in Tomato Plant ABSTRACTSucrose phosphate synthase (SPS) is a key enzyme responsible for sucrose biosynthesis. In its regulation, SPS activity is modulated by an allosteric effector glucose-6-phosphate (G6P) suggested to have an ability to bind SPS N-terminus domain. To understand the role of N-terminus in regulating SPS, the SPS gene was mutated with the deletion of N-terminus domain (∆N-SPS). The ∆N-SPS gen was transformed into tomato plants with 5% transformation efficiency. Three transgenic tomato plant 4.20, 5.5.1, and 5.10 were obtained and confirmed by PCR analysis. Transgenic tomato expression was characterized by enzymatic analysis. Result showed that the G6P allosteric regulation in transgenic ∆N-SPS had lost and the SPS activity increased by 2-fold compared to non-transgenic plant. This showed that N-terminus domain-deleted SPS could be actively expressed in plant. Keywords: enzyme, genetic transformation, N-terminus domain deletion, sucrose phosphate synthase, tomato ABSTRAKSucrose phosphate synthase (SPS) merupakan enzim kunci yang bertanggung jawab dalam sintesis sukrosa. Dalam regulasinya, aktifitas SPS dipengaruhi oleh alosterik efektor glukosa-6-fosfat (G6P) yang diduga dapat berikatan pada domain N-terminus SPS. Untuk mengetahui peran N-terminus pada regulasi SPS, dilakukan mutasi SPS dengan penghilangan domain N-terminus (∆N-SPS). Gen ∆N-SPS diinsersi pada tanaman tomat melalui transformasi genetik dengan efisiensi transformasi 5%. Tiga tanaman transgenik tomat (event4.20; 5.5.1; dan 5.10) didapatkan dan positif terkonfirmasi melalui analisis PCR. Ekspresi mutan dikarakterisasi melalui analisis enzimatik. Hasil menunjukkan bahwa tanaman tomat transgenik ∆N-SPS tidak dipengaruhi regulasi alosterik G6P dan aktifitas SPS 2 kali lipat lebih tinggi daripada tanaman bukan transgenik. Ini menunjukkan bahwa SPS dengan delesi domain N-terminus dapat terekspresi aktif pada tanaman.  Kata Kunci: delesi domain N-terminus, enzim, sucrose phosphate synthase, tomat, transformasi genetik 
TELAAH METODE DIAGNOSIS CEPAT DAN PENGOBATAN INFEKSI Salmonella typhi Hardianto, Dudi
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 6 No. 1 (2019): June 2019
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (542.569 KB) | DOI: 10.29122/jbbi.v6i1.2935

Abstract

Review on Rapid Diagnosis Method and Treatment of Salmonella typhi Infection ABSTRACTSalmonella is a genus of gram-negative bacilli which are pathogenic for human. Recently over 2,500 serotypes of Salmonella have been reported. Of these, the most common serotype causing typhoid fever which is acute infectious disease in small intestine due to S. typhi entering the body through contaminated food or drink. S. typhi infection remains a major public health concern worldwide because of the subsequent economic burden for the cost of surveillance, prevention, and treatment. In Indonesia, typhoid fever is an endemic disease that threatens public health and becomes a complex problem because it increases career cases and drug resistance, so its diagnosis is needed. Although there is already a diagnosis method of typhoid fever conventionally, a fast, easy and reliable diagnosis method is needed to diagnose typhoid fever by medical personnel in endemic countries. Typhoid fever is treated by antibiotics and prevention efforts are carried out through vaccination.Keywords: antibiotics, pathogen, rapid detection, Salmonella typhi, typhoid fever ABSTRAKSalmonella adalah bakteri basil gram negatif yang bersifat patogen terhadap manusia dan saat ini telah dilaporkan lebih dari 2.500 serotipe. Salah satu serotype Salmonella diketahui menyebabkan penyakit demam tifoid yaitu infeksi akut pada usus halus akibat S. typhi yang masuk ke dalam tubuh melalui makanan dan minuman yang tercemar. Infeksi S. typhi menjadi masalah utama dalam kesehatan masyarakat di seluruh dunia karena bebani ekonomi yang ditimbulkannya untuk biaya pengawasan, pencegahan, dan pengobatan. Di Indonesia, demam tifoid merupakan penyakit endemis yang mengancam kesehatan masyarakat dan menjadi masalah kompleks karena demam tifoid meningkatkan kasus-kasus karier dan resistensi obat sehingga diperlukan diagnosisnya. Meskipun sudah ada diagnosis demam tifoid secara konvensional, tetapi diperlukan metode diagnosis yang cepat, mudah dan andal untuk mendeteksi demam tifoid oleh tenaga medis yang bekerja di negara-negara endemik. Demam tifoid diobati dengan pemberian antibiotika dan dilakukan upaya pencegahan melalui vaksinasi.Kata Kunci: antibiotika, demam tifoid, deteksi cepat, patogen, Salmonella typhi
Appendix JBBI Vol 6, No 1, June 2019: Keyword Index and Author Index Sriherwanto, Catur
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 6 No. 1 (2019): June 2019
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (387.513 KB) | DOI: 10.29122/jbbi.v6i1.3692

Abstract

PRODUKSI LIPASE DARI ISOLAT KAPANG HASIL MUTASI UNTUK TRANSESTERIFIKASI Nabilasani, Galih Cendana; Siswodarsono, Trismilah; Suhendar, Dadang; Mubarik, Nisa Rachmania
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 6 No. 1 (2019): June 2019
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (280.35 KB) | DOI: 10.29122/jbbi.v6i1.3047

Abstract

Lipase Production by Mutant Fungal Isolates for Transesterification ABSTRACTLipase is used amongst others in biodiesel production, namely in the transesterification reaction. Kernel B (KB) was a fungus isolated from the waste of palm kernel and seed. The fungus produced lipase that catalysed the transesterification reaction with a lower activity compared to that of AK Amano commercial lipase. The purpose of this study was to obtain mutant fungi with higher transesterification activities than the wild type (KB). The mutation process was carried out using ultraviolet (UV) light, ethyl methane sulfonate (EMS), and N-methyl-N’-nitro-N-nitrosoguanidine (NMNG) on KB fungus. The mutations using UV light produced 11 isolates, of which isolate m4.1KB1 produced a higher transesterification activity (0.172 U·mg-1) compared to the wild type. Mutant m5.7KB, which was generated from mutant m4.1KB1 treated using EMS, had its transesterification activity decreased to only 0.051 U·mg-1. Mutant m6.0,3KB2, which was resulted through NMNG treatment, experienced an increase in transesterification activity which was 91.2% higher than that of KB.Keywords: ethyl methane sulfonate, lipase, mutant fungi, N-methyl-N’-nitro-N-nitrosoguanidine, ultraviolet ABSTRAKLipase dimanfaatkan salah satunya dalam produksi biodiesel, yaitu dalam reaksi transesterifikasi. Kernel B (KB) merupakan kapang yang diisolasi dari limbah inti dan biji kelapa sawit, yang menghasilkan lipase sebagai katalis dalam reaksi transesterifikasi. Namun aktivitas transesterifikasi yang dihasilkan oleh lipase dari KB lebih rendah dibandingkan dengan lipase komersial AK Amano. Tujuan penelitian ini adalah mendapatkan mutan kapang dengan aktivitas transesterifikasi yang lebih tinggi dibandingkan tipe liarnya (KB). Proses mutasi dilakukan dengan menggunakan sinar ultraviolet (UV), ethyl methane sulfonate (EMS), dan N-methyl-N’-nitro-N-nitrosoguanidine (NMNG) terhadap kapang KB. Mutasi KB dengan menggunakan sinar UV menghasilkan 11 isolat, dimana isolat dengan kode m4.1KB1 menghasilkan aktivitas transesterifikasi yang lebih tinggi dibandingkan tipe liar, yaitu 0,172 U·mg-1. Mutan m5.7KB, yang dihasilkan dari mutan m4.1KB1 dengan perlakuan EMS, mengalami penurunan aktivitas transesterifikasi hingga hanya sebesar 0,051 U·mg-1. Mutan m6.0,3KB2 hasil perlakuan NMNG mengalami peningkatan aktivitas transesterifikasi sebesar 91,2% lebih tinggi dari KB.Kata Kunci: ethyl methane sulfonate, kapang mutan, lipase, N-methyl-N’-nitro-N-nitrosoguanidine, ultraviolet
PENGARUH TEKNIK PENGAPUNGAN HAYATI MELALUI FERMENTASI Rhizopus sp. TERHADAP KANDUNGAN NUTRISI PAKAN IKAN APUNG Paramadini, Sabrina Ayu; Sriherwanto, Catur; Nurlaila, .; Suja'i, Imam; Annisa, Mutiara Ayu
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 6 No. 1 (2019): June 2019
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (756.154 KB) | DOI: 10.29122/jbbi.v6i1.3477

Abstract

Influence of Biofloatation Technique through Rhizopus sp. Fermentation on the Nutritional Quality of the Floating Fish FeedABSTRACTSolid fermentation using the mold Rhizopus has been used as an alternative method to improve the physical quality of fish feed, namely stability in water and floatability. Although the fermented fish feed produced had been shown previously to have better stability in water and floatability, the effect of the fermentation on the fish feed nutrition has not yet been known. This study aimed to determine the effect of solid fermentation using the mold Rhizopus sp. on the nutrient content of the fermented feed. In addition, the effect of adding tapioca as much as 0, 1, 2, 3, and 4 g to the dry weight loss and density of the fermented feed was investigated. The results showed that the fermented feed contained higher levels of ash and protein than that before fermentation. The addition of tapioca up to 4 g had no significat effect (p>0.01) on the dry weight loss of the fermented feed, but tended to increase its density compared to those without tapioca addition.Keywords: density, fermentation, fish feed, nutrition, Rhizopus ABSTRAKFermentasi padat menggunakan kapang Rhizopus telah digunakan sebagai metode alternatif untuk memperbaiki kualitas fisik pakan ikan, yakni stabilitas dalam air dan daya apung. Meskipun pakan ikan fermentasi yang dihasilkan sebelumnya sudah dibuktikan memiliki stabilitas dalam air dan daya apung yang lebih baik, namun pengaruh fermentasi terhadap nutrisi pakan ikan tersebut belumlah diketahui. Penelitian ini bertujuan mengetahui pengaruh fermentasi padat menggunakan kapang Rhizopus sp. terhadap kandungan nutrisi pakan fermentasi, dan pengaruh penambahan tapioka sebanyak 0, 1, 2, 3, dan 4 g terhadap kehilangan berat kering dan mass jenis pakan hasil fermentasi. Hasilnya menunjukkan bahwa pakan fermentasi mengandung kadar abu dan protein lebih tinggi dibandingkan sebelum difermentasi. Penambahan tapioka hingga 4 g tidak berpengaruh nyata (p>0,01) pada kehilangan berat kering pakan fermentasi, namun cenderung meningkatkan massa jenis dibandingkan tanpa penambahan tapioka.Kata kunci: fermentasi, massa jenis, nutrisi, pakan ikan, Rhizopus
PENGARUH VARIASI KONSENTRASI METANOL DAN LAMA INDUKSI TERHADAP EKSPRESI PROINSULIN OLEH Pichia pastoris SECARA INTRASELULER Martius, Efrida; Triyadi, Andree; Sofia, Dewi Yustika; Mahsunah, Anis Herliyati
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 6 No. 1 (2019): June 2019
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1800.186 KB) | DOI: 10.29122/jbbi.v6i1.3176

Abstract

The Effects of Variation in Methanol Concentration and Induction Time on Intracellular Proinsulin Expression by Pichia pastoris ABSTRACTDiabetes is a metabolic disorder characterized by hyperglycemia. There were 215 million diabetic patients in 2014 and the number is expected to rise in 2040. Generally, insulin is used to treat diabetic patients. Insulin production by recombinant technology has been done, though still inefficient, by using E. coli and S. cerevisiae expression system. Another alternative expression system is methylotrophic yeast Pichia pastoris. In this research, proinsulin has been expressed by P. pastoris intracellularly. P. pastoris strains used in this research were X33, GS115, and KM71H. All recombinant strains were MutS. Best cultivation media was BMGY. Proinsulin expression was observed at 25°C. Pichia pastoris strain that expressed proinsulin best was GS115-PI. It was supported by PCR in which the strain GS115-PI gave 504 bp-sized bands. Based on proinsulin formation time, the final methanol concentration of 0.5% in 72 hours was found to be the best treatment.Keywords: BMGY, methanol, phenotype, Pichia pastoris, proinsulin ABSTRAKDiabetes melitus merupakan kelainan yang ditandai dengan hiperglikemia. Penderita diabetes pada tahun 2014 di dunia mencapai 215 juta dan diperkirakan akan meningkat pada tahun 2040. Umumnya penderita diabetes diberi pengobatan insulin sehingga menunjukkan akan ada peningkatan kebutuhan insulin. Produksi insulin dengan teknologi DNA rekombinan telah dilakukan dengan menggunakan sistem ekspresi E. coli dan S. cerevisiae namun masih belum efisien. Sistem alternatif lain adalah ragi metilotropik Pichia pastoris. Dalam penelitian ini dilakukan ekspresi proinsulin dari P. pastoris secara intraseluler. Galur P. pastoris yang digunakan dalam penelitian ini adalah X33, GS115, dan KM71H. Semua galur rekombinan adalah MutS. Media tumbuh terbaik adalah BMGY. Ekspresi proinsulin terlihat pada suhu 25°C. Hasil PCR menunjukkan bahwa galur GS115-PI yang dapat menghasilkan pita amplikon berukuran 504 bp. Hasil PCR ini dibuktikan oleh hasil seleksi galur yang menunjukkan bahwa galur GS115-PI dapat mengekspresi proinsulin dibandingkan galur lainnya. Berdasarkan kecepatan pembentukan pita protein proinsulin, variasi konsentrasi akhir metanol 0,5% dengan lama induksi 72 jam merupakan perlakuan terbaik.Kata Kunci: BMGY, fenotipe, metanol, Pichia pastoris, proinsulin

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