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All Journal HAYATI Journal of Biosciences Bioma : Berkala Ilmiah Biologi JURNAL SAINS DAN TEKNOLOGI INDONESIA Jurnal Bioteknologi & Biosains Indonesia (JBBI) Journal of Multidisciplinary Applied Natural Science
Dudi Hardianto, Dudi
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Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Computer Science & IT Earth & Planetary Sciences Education Energy Environmental Science Immunology & microbiology Materials Science & Nanotechnology Mathematics Physics

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PENICILLIN PRODUCTION BY MUTANT OF Penicillium chrysogenum Hardianto, Dudi; ., Suyanto; Prabandari, Erwahyuni Endang; Windriawati, Lira; Marwanta, Edy; ., Tarwadi
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 2 No. 1 (2015): June 2015
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (785.364 KB) | DOI: 10.29122/jbbi.v2i1.530

Abstract

Penisilin adalah antibiotika yang pertama kali ditemukan dan digunakan untuk pengobatan infeksi bakteri. Sejak ditemukan penisilin sebagai antibiotika oleh Alexander Fleming pada tahun 1928, banyak usaha dilakukan untuk meningkatkan produktivitas Penicillium chrysogenum. Pemuliaan galur untuk meningkatkan produksi penisilin dapat menggunakan mutasi acak secara fisika dan kimia. Pada penelitian ini, radiasi sinar ultraviolet digunakan untuk mendapatkan mutan P. chrysogenum. Produksi penisilin ditentukan menggunakan HPLC dan produktivitas mutan dibandingkan dengan induk P. chrysogenum. Mutan M12 menghasilkan penisilin 1,23 kali lebih banyak dibandingkan dengan induk P. chrysogenum.Kata kunci: Penisilin, Penicillium chrysogenum, ultraviolet, mutan, radiasi ABSTRACTPenicillin is the first antibiotic discovered and used for treatment of bacterial infections. Since the discovery of penicillin as antibiotic by Alexander Fleming in 1928, much effort has been invested to improve productivity of Penicillium chrysogenum. Strain improvement to increase the penicillin production can be carried out by physical and chemical random mutation. In this research, ultraviolet irradiation was used to obtain P. chrysogenum mutant. Penicillin production was determined by using HPLC and productivity of P. chrysogenum mutants was compared to the wild type. Mutant M12 produced 1.23 fold higher penicillin than the wild type did.Keywords: Penicillin, Penicillium chrysogenum, ultraviolet, mutant, radiation
BIOKONVERSI SEFALOSPORIN C MENJADI ASAM 7-AMINOSEFALOSPORANAT DENGAN SEFALOSPORIN ASILASE Hardianto, Dudi; Isdiyono, Bima Wedana; Ivan, Fransiskus Xaverius
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 3 No. 2 (2016): December 2016
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (921.244 KB) | DOI: 10.29122/jbbi.v3i2.139

Abstract

Cephalosporins are the most widely used class of β-lactam antibiotic in the world and clinically active against gram positive and gram negative bacteria. Cephalosporin C (CPC) is naturally produced by fungus Cephalosporiun acremonium. CPC has moderate antibacterial activity with minimum inhibitory concentration values of 25-100 µg/mL and 12-25 µg/mL for gram-positive and for gram-negative bacteria, respectively. CPC can be converted into 7-aminocephalosporonic acid (7-ACA) as intermediate compound for cephalosporin derivatives by two-steps or one-step enzymatic method. Two-step enzymatic method uses D-amino acid oxidase (DAAO) to produce glutaryl-7-amino cephalosporanic acid (GL 7-ACA) for the first step and GL-7-ACA acylase to produce 7-ACA for the second step. One-step enzymatic method uses CPC acylase to convert CPC into 7-ACA directly. Some microorganisms produce CPC acylase, such as Pseudomonas sp., Bacillus megaterium, Aeromonas sp., dan Arthrobacler. A natural CPC acylase has low activity and genetic engineering was used to increase its activity.Keywords: Cephalosporin, cephalosporin acylase, 7-ACA, genetic engineering, mutation ABSTRAKSefalosporin merupakan antibiotik golongan β-laktam yang paling banyak digunakan di dunia dan secara klinis aktif terhadap bakteri gram positif dan gram negatif. Sefalosporin C merupakan sefalosporin alami yang dihasilkan oleh kapang Cephalosporium acremonium. Sefalosporin C mempunyai aktivitas antibakteri moderat dengan nilai konsentrasi hambat minimum 25-100 µg/mL untuk bakteri gram positif dan 12-25 µg/mL untuk bakteri gram negatif. Sefalosporin C dapat diubah menjadi asam 7-aminosefalosporanat (7-ACA) sebagai senyawa antara untuk pembuatan turunan sefalosporin dengan metode enzimatik secara dua atau satu tahap. Produksi 7-ACA secara enzimatik dapat menggunakan metode dua tahap dan satu tahap enzimatik. Metode enzimatik secara dua tahap menggunakan enzim asam D-amino oksidase (DAAO) untuk menghasilkan asam glutaril-7-aminosefalosporinat (GL-7-ACA) pada tahap pertama dan menggunakan asam glutaril-7-aminosefalosporinat asilase untuk menghasilkan 7-ACA pada tahap kedua. Metode enzimatik secara satu tahap menggunakan sefalosporin asilase untuk mengubah CPC menjadi 7-ACA secara langsung. Beberapa mikroorganisme penghasil sefalosporin asilase yaitu Pseudomonas sp., Bacillus megaterium, Aeromonas sp., dan Arthrobacter. Aktivitas CPC asilase alami sangat rendah dan rekayasa genetik digunakan untuk meningkatkan aktivitasnya.Kata kunci : Sefalosporin, sefalosporin asilase, 7-ACA, rekayasa genetik, mutasi
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
ANALISA KANDUNGAN ANDROGRAPHOLIDE PADA TANAMAN SAMBILOTO (Andrographis paniculata) DARI 12 LOKASI DI PULAU JAWA Royani, Juwartina Ida; Hardianto, Dudi; Wahyuni, Sri
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 1 No. 1 (2014): December 2014
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (842.933 KB) | DOI: 10.29122/jbbi.v1i1.547

Abstract

Concentration of active compounds contained in medicinal plants is determined by genetic factors as well as growth environment. In sambiloto plants both factors have major impacts on the formation of diterpene lactone, andrographolide. Variation of sampling time, cultivation, and processing methods causes variation in the content of active compounds of the same plant. The purpose of this study was to determine andrographolide concentration of sambiloto plants obtained from 12 different locations with various planting conditions in Java Island. andrographolide content of sambiloto was extracted by methanol and analyzed using HPLC. The results showed that the concentrations of andrographolide varied from 0.29 to 4.44% with an average of 2.19% on dry weight basis. The highest concentration of 4.44% was detected in sambiloto accession from Wonokaton Village, Pasuruan Regency while the lowest one was from Conggeang Kulon Village, Sumedang Regency. Three sambiloto accessions had potential to be further developed as their andrographolide concentrations were above 3%, which was higher than those from all the others.Keywords: Andrographis paniculata, andrographolide, active coumpound, HPLC, Java island ABSTRAKKadar senyawa aktif yang terkandung pada tanaman obat selain dipengaruhi oleh faktor genetik juga dipengaruhi oleh faktor lingkungan tumbuhnya. Pada tanaman sambiloto kedua faktor tersebut berpengaruh sangat besar pada pembentukan diterpen lakton, andrographolide. Adanya variasi pada waktu pengambilan sampel, tempat penanaman, metode pengolahan dan lain sebagainya berakibat pada perbedaan dalam kandungan senyawa aktif pada tanaman yang sama. Tujuan dari penelitian ini adalah untuk mengetahui kadar andrographolide dari tanaman sambiloto yang diambil dari 12 lokasi tumbuh dengan kondisi penanaman yang berbeda di Pulau Jawa. Daun tanaman sambiloto diekstrak dengan methanol kemudian dianalisis kandungan andrographolide menggunakan HPLC. Kadar andrographolide yang dihasilkan bervariasi berkisar antara 0,29-4,44% dengan kadar rata-rata adalah 2,19% berat kering. Kadar tertinggi didapatkan pada aksesi dari Desa Wonokaton Kabupaten Pasuruan dengan kadar andrographolide adalah 4,44% sedangkan kadar yang terendah didapatkan pada aksesi dari Desa Conggeang Kulon, Kab. Sumedang. Berdasarkan data kandungan andrographolide, diperoleh 3 aksesi sambiloto yang potensial untuk dikembangkan menjadi aksesi unggulan karena kadar andrographolidenya di atas 3%, melebihi semua yang lain.Kata kunci: Andrographis paniculata, andrographolide, senyawa aktif, HPLC, pulau Jawa
OPTIMASI METODE LISIS ALKALI UNTUK MENINGKATKAN KONSENTRASI PLASMID Hardianto, Dudi; Indarto, Alfik; Sasongko, Nurtjahjo Dwi
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 2 No. 2 (2015): December 2015
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (635.116 KB) | DOI: 10.29122/jbbi.v2i2.510

Abstract

Plasmids are extra chromosomal molecules of DNA that replicate autonomously and found in prokaryote and eukaryote cells. There are a number of methods that are used to isolate plasmids, such as alkaline lysis, boiling lysis, using cesium chloride, and chromatography. Amongst the disadvantages in plasmid isolation methods are lengthy time especially when handling a large number of samples, high cost, and low purity. Alkaline lysis is the most popular for plasmid isolation because of its simplicity, relatively low cost, and reproducibility. This method can be accomplished in 50 minutes to one hour. In this research, the alkaline lysis method was developed to obtain suitable plasmid for applications in a molecular biology laboratory. The aim of this research was to reduce contaminants and improve yield of plasmid. The result of isolation of pICZA plasmid in Escherichia coli gave the concentration of 3.3 to 3.8 µg/µL with the purity of 1.99.Keywords: Plasmid isolation, pICZ A, Escherichia coli, rapid, alkaline lysis  ABSTRAKPlasmid merupakan molekul DNA ekstrakromosomal yang bereplikasi secara mandiri dan ditemukan dalam sel prokariot dan eukariot. Banyak metode yang digunakan untuk isolasi plasmid, seperti: lisis alkali, lisis dengan pemanasan, penggunaan sesium klorida, dan kromatografi. Kelemahan beberapa metode isolasi DNA adalah waktu isolasi yang lama terutama saat isolasi plasmid dalam jumlah banyak, mahal dan kemurniannya yang rendah. Metode lisis alkali merupakan metode yang sangat umum untuk isolasi plasmid karena mudah dilakukan, relatif murah, dan reprodusibilitas. Metode ini dapat dilakukan dalam 50 menit sampai 1 jam. Pada penelitian ini dikembangkan metode lisis alkali untuk memperoleh plasmid yang sesuai untuk penggunaan di laboratorium biologi molekuler. Tujuan dari penelitian ini adalah untuk mengurangi jumlah kontaminan dan meningkatkan konsentrasi plasmid. Hasil isolasi plasmid pICZA dalam Escherichia coli mempunyai konsentrasi antara 3,3 sampai 3,8 µg/µL dan kemurniannya 1,99.Kata Kunci: Isolasi plasmid, pICZ A, Escherichia coli, cepat, lisis alkali
PRODUKSI REKOMBINAN SEFALOSPORIN ASILASE SEBAGAI BIOKATALIS UNTUK PRODUKSI ASAM 7-AMINOSEFALOSPORANAT Isdiyono, Bima Wedana; Hardianto, Dudi; Ivan, Fransiskus Xaverius
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 4 No. 1 (2017): June 2017
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1087.024 KB) | DOI: 10.29122/jbbi.v4i1.2059

Abstract

Production of Cephalosporin Acylase Recombinant as Biocatalyst for 7-Aminocephalosporanic Acid Production7-aminocephalosporanic acid (7-ACA) is a precursor for the production of semisynthetic cephalosporin derivatives. The enzymatic 7-ACA production can use two-stage and one-step enzymatic methods. Two-stage enzymatic method uses D-amino acid oxidase (DAAO) enzyme to produce glutaryl-7-aminocephalosporanic acid (GL-7-ACA) in the first stage and glutaryl-7-aminocephalosporanic acid acylase to produce 7-ACA in the second stage. The one-stage enzymatic method using cephalosporin acylase (CPC acylase) converts the CPC to 7-ACA directly. The aim of this research was to produce recombinant CPC acylase in Escherichia coli BL21(DE3). Transformantion culture E. coli BL21(DE3) was induced with concentrations of IPTG 0; 0.25; 0.5; 0.75; 1; 2 mM for 5 hours. The induction time of IPTG was determined at 0, 1, 2, 3, 4, and 5 hours. The results showed that CPC acylase produced by E. coli BL21(DE3) with optimum condition of CPC acylase production was 0.5 mM IPTG and optimal induction time of IPTG was 5 hours.Keywords: Cephalosporin, cephalosporin acylase, 7-ACA, protein expression, Escherichia coli BL21(DE3) ABSTRAKAsam 7-aminosefalosporanat (7-ACA) merupakan prekursor untuk produksi turunan sefalosporin semisintetik. Produksi 7-ACA secara enzimatik dapat menggunakan metode dua tahap dan satu tahap enzimatik. Metode enzimatik secara dua tahap menggunakan enzim asam D-amino oksidase (DAAO) untuk menghasilkan asam glutaril-7-aminosefalosporinat (GL-7-ACA) pada tahap pertama dan menggunakan asam glutaril-7-aminosefalosporinat asilase untuk menghasilkan 7-ACA pada tahap kedua. Metode enzimatik satu tahap dengan sefalosporin asilase (CPC asilase) mengubah CPC menjadi 7-ACA secara langsung. Tujuan penelitian adalah memproduksi rekombinan CPC asilase di dalam sel Escherichia coli BL21(DE3). Kultur Transforman E. coli BL21(DE3) diinduksi dengan konsentrasi IPTG 0; 0,25; 0,5; 0,75; 1; 2 mM selama 5 jam. Waktu induksi IPTG ditentukan pada 0, 1, 2, 3, 4 dan 5 jam. Hasil penelitian menunjukan bahwa CPC asilase diproduksi oleh E. coli BL21(DE3) dengan kondisi optimal produksi CPC asilase adalah konsentrasi IPTG 0,5 mM dan waktu induksi IPTG optimal adalah 5 jam.
Cloning and Extracellular Expression of Glargine in Pichia pastoris Hardianto, Dudi; Martius, Efrida; Rostinawati, Tina; Safarrida, Anna; Royani, Juwartina Ida; Assyifa, Fahroziah; Laziba, Dihan
HAYATI Journal of Biosciences Vol. 31 No. 2 (2024): March 2024
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.31.2.248-255

Abstract

Patients with diabetes mellitus increase significantly every year. The increasing number of people with diabetes mellitus results in increased insulin requirements. There are two types of insulin used for diabetes mellitus treatment: human insulin and insulin analogues. Escherichia coli, Pichia pastoris, Saccharomyces cerevisiae, or Hansenula polymorpaha has been used to produce human insulin and insulin analogues. Pichia pastoris can produce glargine in large quantities, and the insulin protein produced will be secreted outside the cell to facilitate the purification process. The advantage of glargine has a long working time of up to 24 hours. Hence, glargine is more effective because patients with diabetes receive glargine injections only once daily. The research started with cloning the glargine gene, transforming pPICZαA-G plasmid into Pichia pastoris, and testing glargine production. 20 recombinant Pichia pastoris colonies were selected and regenerated. Eight recombinant Pichia pastoris colonies were tested for glargine production, and six colonies were detected producing glargine by electrophoresis SDS-PAGE gel stained with Coomassie blue. This study aims to produce glargine using Pichia pastoris as an expression system capable of producing glargine extracellularly, thus simplifying the purification process.
Modification of a Gene Cassette to Express Proglargine in Pichia pastoris: Elimination of Glu-Ala Repeats, Spacer and C-peptide Sequences Nasution, Uli Julia; Astuti, Rika Indri; Wahyudi, Aris Tri; Hardianto, Dudi; Martius, Efrida
Journal of Multidisciplinary Applied Natural Science Articles in Press
Publisher : Pandawa Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.47352/jmans.2774-3047.368

Abstract

The growing global prevalence of diabetes mellitus has sharply increased the demand for insulin and its analogues. Pichia pastoris is a well-established system for recombinant pro-insulin and its analogues production. However, conventional gene constructs often include additional sequences, such as Glu-Ala repeats, spacer peptides, and c-peptides that complicate downstream processing and reduce efficiency. This study aimed to construct and express a proglargine (PG) gene cassette lacking the Glu-Ala repeats, spacer, and c-peptide in P. pastoris GS115 to obtain a uniform PG protein. The recombinant vector propagated in Escherichia coli TOP10F’, then expressed in P. pastorisGS115. Selected transformants were cultivated in YPG medium, then induced with 1% and 2% methanol daily in BMMY. The optimum methanol concentration further evaluated in ½ BSM induction medium. The result demonstrated that optimal PG expression was achieved with 2% methanol induction in BMMY, producing higher levels than those with ½ BSM. Among the transformants, PG.c2 produced the highest PG protein in BMMY medium induced with 2% methanol. Dot-blot analysis confirmed the secretion of PG, and LC-HRMS analysis demonstrated 100% amino acid sequence coverage of PG, confirming the integrity and completeness of the expressed protein. This study presented a newly modified PG gene cassette, inserted into pPICZαA vector, to express uniform secreted PG in P. pastoris GS115. By simplifying the precursor structure, a more homogeneous precursor product can be obtained, which is expected to simplify purification and also the downstream enzymatic process of PG into mature insulin glargine.
Co-Authors Alfik Indarto, Alfik Amir Musadad Anna Safarrida, Anna Anto Budiharjo Aris Tri Wahyudi Assyifa, Fahroziah Bima Wedana Isdiyono, Bima Wedana Dihan Laziba Edy Marwanta, Edy Efrida Martius ERWAHYUNI ENDANG PRABANDARI Fransiskus Xaverius Ivan, Fransiskus Xaverius gusnidar, tutus Hermin Pancasakti Kusumaningrum Juwartina Ida Royani, Juwartina Ida Lira Windriawati, Lira Marlia Singgih Nasution, Uli Julia Nurtjahjo Dwi Sasongko Rika Indri Astuti Risma Wiharyanti Sri Wahyuni Suyanto . Tarwadi ., Tarwadi Tina Rostinawati Wahono Sumaryono