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Constitutive Expression of Candida antarctica Lipase B (CALB) in Pichia pastoris Using pGAPZα Vector Febriana Dwi Wahyuni; Asrul Muhamad Fuad; Suharsono Suharsono
ANNALES BOGORIENSES Vol 20, No 1 (2016): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (461.586 KB) | DOI: 10.14203/ann.bogor.2016.v20.n1.31-38

Abstract

The CalBsyn gene was previously constructed synthetically to encode Candida antarctica lipase B (CALB). Lipase from CalBsyn gene is slightly different from that of wild type CALB (CALB-wt) where it has three amino acids substitutions at different positions, i.e. V210I, A281E, and V221D, to improve enzyme’s thermostability and catalytic efficiency. The CalBsyn gene was isolated from pJ912-CalBsyn vector by digestion using XhoI restriction enzyme. The CalBsyn gene then was ligated to pGAPZα expression vector and transformed into E. coli TOP10F’ in order to obtain recombinant vector pGAPZα-CalBsyn. The result showed that pGAPZα-CalBsyn recombinant vector was successfully transformed into E. coli TOP10F’ with transformation efficiency of 4.11 x 103 cfu/µg plasmid DNA. The pGAPZα-CalBsyn recombinant plasmid was successfully introduced into Pichia pastoris SMD1168H using electroporation method with transformation efficiency of 1.01 x 102 cfu/µg DNA. Qualitative lipase activity assays showed that transformed P. pastoris secreted recombinant lipase (CALB) and has lipolytic activity; while quantitative lipase activity assays showed that the lipase activity was 63.5 Units/ml in 48 hours. Analysis using SDS-PAGE showed that CALB protein was expressed successfully and the recombinant protein’s molecular size was approximately 45 kDa.
The dynamics of Wnt-5a and Shh expression in tissue regeneration process of mice (Mus musculus) digit tip Titta Novianti; Febriana Dwi Wahyuni; Syafruddin Ilyas
Biogenesis: Jurnal Ilmiah Biologi Vol 10 No 1 (2022)
Publisher : Department of Biology, Faculty of Sci and Tech, Universitas Islam Negeri Alauddin Makassar

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24252/bio.v10i1.27378

Abstract

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TRANSFORMASI PLASMID REKOMBINAN pRI_101-AN MEMBAWA SISIPAN GEN cryIII MELALUI Agrobacterium tumefaciens Febriana Dwi Wahyuni; Muhamad Amza Sidiq; Seprianto Seprianto; Henny Saraswati
BERITA BIOLOGI Vol 21, No 1 (2022)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/beritabiologi.v21i3.4256

Abstract

Sweet potato is a plant that give many benefits because of its high carbohydrate content and is an alternative food source. Behind the benefits of sweet potatoes, there is a threat of pest attack that can cause a decrease in production of up to 19.91%. Sweet potato weevil attack on the leaves, stalks, stems, and tubers. The solution to against this pest attack is by engineering using the cry III gene mediated by A. tumefaciens. The aim of this study was to transform a recombinant plasmid carrying the insertion of the cry III gene into A. tumefaciens cells. The results showed that the recombinant plasmid pRI_101-AN carrying the insertion of the cry III gene was successfully transformed into A. tumefaciens cells by electroporation method by producing 36 recombinant clones on LB media with neomycin and kanamycin antibiotics. Results of colony PCR, DNA bands measuring 1900 bp were successfully amplified using gene-specific primers. Results of the isolation of plasmid DNA bands measuring 12000 bp were obtained from the results of electrophoresis. This size corresponds to the recombinant plasmid pRI_101-AN-cry III.
Optimization of Real-Time PCR Conditions for COVID-19 Diagnosis with Logix Smart Reagent™ Anisa Febriyanti; Seprianto Seprianto; Titta Novianti; Febriana Dwi Wahyuni; Oktaviani Naulita Turnip; Roselein Putri; Henny Saraswati
BIOEDUKASI Vol 20 No 1 (2022)
Publisher : PROGRAM STUDI PENDIDIKAN BIOLOGI FAKULTAS KEGURUAN DAN ILMU PENDIDIKAN UNIVERSITAS JEMBER

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.19184/bioedu.v20i1.28356

Abstract

Corona Virus Disease 2019 or COVID-19 is a new type of virus that attacks the respiratory tract and can cause death. Laboratory examinations play an essential role in diagnosing COVID-19 with a set of reagents or kits. Sampiand sampling is carried out with a nasopharyngeal swab or oropharyngeal swab. Positive samples of COVID-19 patients used in this study were converted into RNA at the COVID-19 Referral Clinic in Bekasi, after which volume optimization was carried out with a total volume of 5 µl, 8 µl, and 10 µl with the Logix Smart™ kit. The method in this study uses One-Step Real-time PCR. This method is the best method for carrying out several bear tests because it can reduce the possibility of sample contamination. The procedure is fast and has high sensitivity. The fluorescence detection used in this study was FAM with a specific target of COVID-19 RNA and ROX with a particular DNA target of RNase-P. This research was conducted to obtain optimal volume conditions under the manufacturer's standards in detecting the SARS-CoV-2 virus. The results of this study indicate that a total volume of 5 l is the optimal total volume for detecting the presence of the SARS- CoV-2 virus in samples taken from patients.
UJI AKTIVITAS ANTIOKSIDAN DENGAN METODE DPPH DAN IDENTIFIKASI GOLONGAN METABOLIT SEKUNDER PADA DAGING UBI JALAR DARI BERBAGAI DAERAH DI INDONESIA: ANTIOXIDANT ACTIVITY TEST USING DPPH METHOD AND IDENTIFICATION OF SECONDARY METABOLITES IN SWEET POTATOES FROM VARIOUS REGIONS IN INDONESIA Meisya Then Septian; Febriana Dwi Wahyuni; Adri Nora
SPIN JURNAL KIMIA & PENDIDIKAN KIMIA Vol. 4 No. 2 (2022): Juli - Desember 2022
Publisher : UIN Mataram

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20414/spin.v4i2.5734

Abstract

Antioksidan dapat ditemukan di beberapa bahan pangan salah satunya ubi jalar. Ubi jalar telah dikonsumsi sebagai bahan pangan di beberapa daerah di Indonesia seperti Papua dan Maluku. Ubi jalar yang digunakan dalam penelitian ini diambil dari beberapa daerah yaitu Riau, Tomohon, Balikpapan, Jambi, Malang, Pontianak, Kupang, Bangka, Medan, dan Merauke. Sampel ubi jalar dari berbagai daerah dan kondisi lingkungan yang berbeda, membuat ubi jalar diprediksi memiliki kandungan senyawa metabolit sekunder yang berbeda. Pada penelitian ini dilakukan pengujian antioksidan dengan metode 2,2-difenil-1-pikrilhidrazil (DPPH), uji total fenol, dan pengujian fitokimia. Hasil yang didapatkan menunjukkan bahwa aktivitas antioksidan yang paling tinggi terdapat pada ubi jalar Medan berdaging oranye tua dengan nilai IC50 sebesar 235,34 mg/mL. Sementara total fenol yang paling tinggi terdapat pada ubi jalar Medan berdaging ungu sebesar 6,035 ?g GAE/g. Pada pengujian fitokimia didapatkan tidak semua ubi mengandung metabolit sekunder yang sama. Pada pengujian alkaloid dan steroid semua sampel ubi jalar tidak mengandung senyawa golongan alkaloid dan steroid. Pengujian flavonoid pada sampel daging ubi jalar dari Tomohon, Balikpapan, Jambi, Malang, Pontianak (daging oranye tua), Kupang, Bangka, dan Medan (daging oranye tua dan oranye muda) mengandung senyawa golongan flavonoid. Sementara pada uji terpenoid sampel daging ubi jalar dari Riau, Tomohon, Balikpapan, Malang, Kupang, Bangka, dan Medan (daging ungu) mengandung senyawa golongan terpenoid.
UJI AKTIVITAS ANTIBAKTERI EKSTRAK METANOL UBI JALAR (IPOMOEA BATATAS L.) TERHADAP PERTUMBUHAN Escherichia coli dan Staphylococcus aureus Atikah Fairus Siti Badriah; Febriana Dwi Wahyuni; Adri Nora
AL-ULUM: JURNAL SAINS DAN TEKNOLOGI Vol 8, No 1 (2022)
Publisher : Universitas Islam Kalimantan Muhammad Arsyad Al Banjari

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.31602/ajst.v8i1.7180

Abstract

Antibacterial is a compound capable of inhibiting and killing pathogenic bacteria. Escherichia coli and Staphylococcus aureus are pathogenic bacteria because they can cause disease if infected. Sweet potato is an alternative food that is often consumed by some Indonesian people and is thought to contain antibacterial compounds. The sweet potatoes used in this study came from Riau (A), Tomohon (B), Balikpapan (C), Jambi (D), Malang (E), Pontianak (F), Kupang (G), Bangka Belitung (H) , Medan (I), Balikpapan (J), and Merauke (K). Antibacterial testing was carried out using the disc diffusion method and using Eosin Methylene Blue (EMB) and Luria Bertani (LB) media with sample concentrations of 250, 500, and 700 ppm. The study was conducted using the antimicrobial activity testing method which was carried out only once in an experiment and observations would be made after 24 hours of incubation. From the results obtained, the optimum growth of E. Coli at a concentration of 700 ppm with samples from the Kupang area (G) with purple color with an inhibition zone of 8.5 mm and a minimum at a concentration of 250 ppm with samples originating from the Merauke (K) area with white color with a white zone. resistance of 1 mm. Meanwhile, the test results on the optimum growth of S. aureus at a concentration of 700 ppm with samples from West Kalimantan (F2) with purple color with an inhibition zone of 7 mm and a minimum at a concentration of 250 ppm with samples from Balikpapan (C), Bangka Belitung. (H), and Medan (I3) are white and light orange with an inhibition zone of 2 mm.
PEMANFAATAN SAMPAH ORGANIK RUMAH TANGGA MENJADI ECO-ENZYME CAIRAN SEJUTA MANFAAT DI CLUSTER MALTA SENTRALAND PARADISE KEC. PARUNG PANJANG Seprianto Seprianto; Henny Saraswati; Febriana Dwi Wahyuni; Titta Novianti; Adri Nora; Putri Handayani
J-ABDI: Jurnal Pengabdian kepada Masyarakat Vol. 2 No. 8: Januari 2023
Publisher : Bajang Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.53625/jabdi.v2i8.4528

Abstract

Keberadaan sampah dari limbah rumah tangga yang berlebihan di lingkungan merupakan salah satu masalah penting karena dapat merusak keseimbangan ekosistem lingkungan. Perlu adanya pengelolaan terhadap limbah rumah tangga tersebut supaya tidak menimbulkan dampak negatif bagi lingkungan dan kesehatan masyarakat. Perumahan Sentraland paradise merupakan perumahan baru yang terletak di Desa Kabasiran, Kecamatan Parung Panjang Kab. Bogor yang terdiri berbagai cluster salah satunya cluster Malta. Salah satu pemanafaatan sampah organik dari limbah rumah tangga adalan cairan serbaguna eco enzyme. Cairan serbaguna ini bisa menjadi karbol pembersih alami, sabun cuci piring, pemurni udara, pembersih luka, sanitizer alami, pupuk cair, dan cairan pestisidia untuk tanaman. Ada 10 warga yang ikut dalam kegiatan ini, sedikitnya warga yang ikut dikarenakan punya kegiatan lain sehingga tidak dapat berpartisipasi. Selama kegiatan berlangsung peserta sangat aktif bertanya dan berdiskusi dengan pemateri. Pelaksanaan praktek warga ikut terlibat dalam pembuatan eco-enzyme. Kegiatan ini sangat bermanfaat karena banyak informasi yang didapatkan warga tentang bagaimana pemanfaatan sampah dapur yang selama ini hanya dibuang menjadi produk yang bernilai. Kegiatan ini diharapkan dapat memberikan solusi dalam penanganan sampah organik rumah tangga menjadi produk eco-enzyme yang memiliki sejuta manfaat serta menjadi produk komersial yang dapat menjadi tambahan pendapatan warga Malta.
Optimasi Volume Kit Da An Gene Untuk Deteksi SARS-CoV-2 dengan Real Time RT-PCR Seprianto; Muhammad Arreza; Titta Novianti; Febriana Dwi Wahyuni; Oktaviani Naulita Turnip; Roaslein Putri; Henny Saraswati
BIOEDUSCIENCE Vol 6 No 2 (2022): BIOEDUSCIENCE
Publisher : Universitas Muhammadiyah Prof. Dr. Hamka

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (566.732 KB) | DOI: 10.22236/j.bes/628595

Abstract

Background: SARS-CoV-2 is a new type of coronavirus of the genus Betacoronavirus and the family Coronaviridae that causes a respiratory disease called COVID-19. The virus has a sheath and genetic material in the form of single-chain RNA. The genome structure of this virus is divided into two types, namely genes that encode non-structural proteins consisting of the ORF1a / ORF1b gene and genes that encode structural proteins consisting of spike glycoprotein (S), envelope (E), membrane glycoprotein (M), and nucleocapsid protein (N). Methods: The method of detecting SARS-CoV-2 with real time RT-PCR is the most recommended method because it has high specificity and accuracy. The specificity of a method is necessary to be able to specifically recognize the pathogen that causes the disease. Real time RT-PCR requires sampling with a swab on the oropharynx or nasopharynx to be examined in the laboratory which later the presence of viral RNA becomes a molecule that is assessed for diagnosis results. In this study, volume optimization was carried out on the Da An Gene kit used for the detection of SARS-CoV-2 with Reverse Transcription Polymerase Chain Reaction (Real time RT-PCR) with the aim of saving the use of reagents from available kits but with amplification results remaining optimal and accurate. Results: There were three SARS-CoV-2 RNA samples used consisting of N62, N63, and N79 samples and three types of total volume used were 20 μl, 15 μl, and 10 μl. The results of this study showed that the three positive samples contained SARS-CoV-2 with a Cq value of < 40. Conclusion: A volume of 20 μl is the optimal volume, which is more efficient than the manufacturer's recommended volume of 25 ul.
MUTATION DETECTION OF MULTIDRUG-RESISTANT TUBERCULOSIS BY RT-PCR METHOD AS THE DIAGNOSTIC TOOL OF MDR-TB Titta Novianti; alfero Putra Iryanto; feby feby; callista marsya; putri mega utami; febriana dwi wahyuni; henny saraswati; seprianto; adri nora; roaslein putri; nie nie; sabar pambudi
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 10 No. 1 (2023): Juni 2023
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29122/jbbi.v10i1.5653

Abstract

Eight percent of tuberculosis (TB) cases worldwide are resistant to rifampicin, with mutations occurring in the rpoB and katG genes. It is necessary to develop a specific multidrug-resistant (MDR) diagnostic technique using the RT-PCR method in Indonesia to aid in rapid and accurate diagnosis. In-silico testing using SnapGene software resulted in the design of DNA primers for the katG and rpoB genes, plasmids, and specific probes. This study employed a cross-sectional design using 30 non-MDR-TB and MDR-TB samples from RSUD Sitanala, Tangerang Banten, which were tested for amplification of the katG and rpoB genes using Sybr green RT-PCR. Validity testing was conducted using specific probes for the katG and rpoB genes. The amplification results showed that MDR-TB samples and MDR-TB plasmids required a longer time compared to non-MDR-TB samples and non-MDR-TB plasmids. The Quantification Cycle (Cq) value in non-MDR-TB samples was lower than the Cq value in MDR-TB samples. A t-test revealed a significant difference in Cq values of the rpoB and katG genes between MDR-TB and non-MDR-TB patients (p-value < 0.005). These differences in Cq values indicate that the findings of this study can serve as an initial reference for the development of an RT-PCR-based diagnostic kit for MDR-TB.
Optimasi Volume Kit Da An Gene Untuk Deteksi SARS-CoV-2 dengan Real Time RT-PCR Seprianto Seprianto; Muhammad Arreza; Titta Novianti; Febriana Dwi Wahyuni; Oktaviani Naulita Turnip; Roaslein Putri; Henny Saraswati
BIOEDUSCIENCE Vol 6 No 2 (2022): BIOEDUSCIENCE
Publisher : Universitas Muhammadiyah Prof. Dr. Hamka

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22236/j.bes/628595

Abstract

Background: SARS-CoV-2 is a new type of coronavirus of the genus Betacoronavirus and the family Coronaviridae that causes a respiratory disease called COVID-19. The virus has a sheath and genetic material in the form of single-chain RNA. The genome structure of this virus is divided into two types, namely genes that encode non-structural proteins consisting of the ORF1a / ORF1b gene and genes that encode structural proteins consisting of spike glycoprotein (S), envelope (E), membrane glycoprotein (M), and nucleocapsid protein (N). Methods: The method of detecting SARS-CoV-2 with real time RT-PCR is the most recommended method because it has high specificity and accuracy. The specificity of a method is necessary to be able to specifically recognize the pathogen that causes the disease. Real time RT-PCR requires sampling with a swab on the oropharynx or nasopharynx to be examined in the laboratory which later the presence of viral RNA becomes a molecule that is assessed for diagnosis results. In this study, volume optimization was carried out on the Da An Gene kit used for the detection of SARS-CoV-2 with Reverse Transcription Polymerase Chain Reaction (Real time RT-PCR) with the aim of saving the use of reagents from available kits but with amplification results remaining optimal and accurate. Results: There were three SARS-CoV-2 RNA samples used consisting of N62, N63, and N79 samples and three types of total volume used were 20 μl, 15 μl, and 10 μl. The results of this study showed that the three positive samples contained SARS-CoV-2 with a Cq value of < 40. Conclusion: A volume of 20 μl is the optimal volume, which is more efficient than the manufacturer's recommended volume of 25 ul.