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Isolation and Screening of Lactic Acid Bacteria From Sumbawa Buffalo Milk (Bubalus bubalis) as Potential Starter Cultures Harmoko, Deni; Ardyati, Tri; Jatmiko, Yoga Dwi
The Journal of Experimental Life Science Vol. 12 No. 3 (2022)
Publisher : Graduate School, Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jels.2022.012.03.03

Abstract

The Sumbawa buffalo (Bubalus bubalis) is one of the ruminant livestock in Indonesia that not only contributes to fulfilling meat requirements but also milk. Besides containing nutrients that are very beneficial for human health, buffalo milk is also a potential source of lactic acid bacteria (LAB) with technological and functional properties. Lactic acid bacteria have been utilized as starter cultures in various fermented products. This study aimed to isolate LAB from Sumbawa buffalo milk and to identify the potential isolate as a starter culture. The screening of LAB as a starter culture was based on some technological properties, including proteolytic activity, lipolytic activity, exopolysaccharide (EPS) production, antibacterial activity, antibiotic sensitivity, hemolytic activity, and acidification activity Data were analyzed statistically using one-way ANOVA and Tukey’s post hoc test at a 5% significance level. A total of 21 isolates were isolated from fresh buffalo milk, with a LAB total was 4.7x105 CFU.mL-1. All the isolates were characterized as Gram-positive with cocci-shaped. The SA8 isolate was selected as the most potential candidate as a starter culture because it has fulfilled the criteria such as the highest proteolytic activity, the lowest lipolytic activity, producing EPS, potential antagonistic activity against Bacillus cereus, Escherichia coli, and Salmonella Typhi, and sensitivity to cefazolin, intermediate to erythromycin and cinoxacin, non-pathogen, as well as the most rapid acidification activity. The SA8 isolate was identified as Enterococcus lactis with a similarity level of 99.99% towards strain BT159. This indigenous LAB was a potential starter culture of Sumbawa fermented buffalo milk to increase the diversification of products derived from buffalo milk. Keywords: Enterococcus lactis, lactic acid bacteria, technological properties, starter culture, Sumbawa buffalo milk.
Halophilic Bacteria Producing Protease from Salted Fish in Ponrang, Luwu Regency, South Sulawesi Ramadhan, Andi Muhammad Faiz; Ardyati, Tri; Jatmiko, Yoga Dwi
The Journal of Experimental Life Science Vol. 13 No. 1 (2023)
Publisher : Graduate School, Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jels.2023.013.01.06

Abstract

The need for protease enzymes for medical and industrial purposes. The need for proteases in the world reaches 65% of the total sales of enzymes, and in Indonesia can reach 2.500 tons every year, and 99% percent of the enzyme needs are still imported from abroad. Salted fish is one of the foods that contain a lot of protein, which is about 42% in 100 g of salted fish. It allows the presence of proteolytic bacteria that have halophilic properties in salted fish. This study aims to explore the presence of proteolytic bacteria with halophilic properties in salted fish. Proteolytic isolates were isolated using SMA media from salted fish from Ponrang District, Luwu Regency, South Sulawesi. A qualitative test was carried out by measuring the clear zone formed in Skim Milk Agar (SMA). From the isolation process, 51 isolates were obtained. However, after the screening, only 25 pure isolates were found that formed a clear zone, followed by a quantitative test to see which isolates had stable activity at incubation times of 24, 48, and 72 hours using Tryptic Soy Broth (TSB) media. The results obtained four superior isolates, P1A1K, P2B2PS, P3C6PS, and P3C6P, then continued to quantify halophilic bacteria properties by administering NaCl with 15% and 20% concentration into TSB media for 24 and 48 hours incubation. Two isolates with the highest protease activity were P3C6P isolates with the activity of 43.23 ± 7.11 U.mL-1 at 15% salt concentration and 42.83 ± 3.04 U.mL-1 at 20% salt concentration and P2B2PS isolates of 38.05 ± 4.05 U.mL-1 at 15% salt concentration and 38.15 ± 1.47 U.mL-1 at 20% salt concentration. The two isolates were then tested for pathogenicity on blood agar media. It was found that only P3C6P isolates did not have pathogenic properties, so P3C6P isolates were continued with catalase, oxidase, and gram staining tests, which showed negative catalase and oxidase results and were gram-positive, followed by identification based on sequences. 16S rDNA and phylogenetic tree construction where isolate P3C6P was identified as Bacillus cereus with a 100% similarity level to the WHX1 strain. Keywords: Enzyme, Protease, Proteolytic, Salted Fish, 16s-rDNA.
Effect of Combination of Different Antibiotics and Promoters for Expressing Recombinant Darbepoetin in Stable CHO K-1 Cell Line: Evaluating Antibiotic Combinations to Improve Yield and Quality of Darbepoetin Widekdo, Dwi purno; Widodo, Nashi; Rifa'i, Muhaimin; Jatmiko, Yoga Dwi
Journal of Tropical Life Science Vol. 15 No. 1 (2025)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/

Abstract

Antibiotics are key for successful molecular cloning techniques. Different antibiotics have different mechanisms of action, which leads to cell heath and a viable number of passages. Moreover, the suitability of the promoter also plays an important role in achieving a higher level of protein titter in the stable cell platform. Therefore, with plenty of options of antibiotics and promoters available, we need to determine the best combination of antibiotics and promoters, particularly for specific proteins of interest. Darbepoetin is a recombinant therapeutics protein with extra glycosylation to increase the half-life in the blood; this drug is used for the administration of CKD and leukemia patients. Blasticidin-S and puromycin were used as antibiotics, and CMV and EF-1 promoters were used in this experiment to evaluate the expression of recombinant darbepoetin for the protein model. CHO K-1 cell line was transfected with a plasmid carrying a combination of promoter and antibiotics genes; after 14 days, the level of specific protein expression was evaluated using the western blot technique. A single clone cell was obtained using the serial dilution method to evaluate the clonality and expression of the protein of interest. This study successfully obtained a single clone from stable pool transfection. This result suggested that a combination of puromycin antibiotics and EF-1 promoter has promising expression compared to Blasticidin-S antibiotics with CMV promoter. For further conclusion, an analytical comparison of both combinations needed to be done.
Uji Aktivitas Bakteriofage Litik dari Limbah Rumah Tangga Terhadap Salmonella Typhi Jatmiko, Yoga Dwi; Purwanto, Agung Putra; Ardyati, Tri
Jurnal Biodjati Vol 3 No 2 (2018): November
Publisher : UIN Sunan Gunung Djati Bandung

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15575/biodjati.v3i2.3471

Abstract

Salmonella Typhi merupakan salah satu bakteri yang menjadi agen penyakit bawaan makanan. Bakteriofage sebagai alternatif penggunaan antibiotika telah digunakan untuk mengendalikan bakteri tersebut. Penelitian ini dilakukan untuk mendapatkan isolat bakteriofage litik yang mampu melisis beberapa bakteri patogen yang diujikan dan mengetahui pengaruh aktivitas bakteriofage litik terhadap pertumbuhan SalmonellaTyphi. Bakteriofage diisolasi dari limbah rumah tangga. Selanjutnya penentuan host range bakteriofage terhadap bakteri patogen lain dilakukan dengan metode spot test. Uji aktivitas bakteriofage terhadap SalmonellaTyphi dilakukan menggunakan metode bacterial challenge test. Berdasarkan hasil isolasi, didapat enam isolat bakteriofage, yaitu B2-St, B3-St, S1-St, S2-St, SL1-St, dan SL3-St. Semua isolat bakteriofage mampu melisiskan sel bakteri Escherichia coli dan Salmonella Typhimurium namun tidak mampu melisiskan Bacillus cereus, Staphylococcus aureus dan Shigella disentriae.Tiga isolat bakteriofagetelah terpilih berdasarkan densitas plaque terbanyak yaitu B2-St, SL3-St dan S2-St. Kemampuan isolat bakteriofage B2-St dalam melisiskan sel Salmonella Typhi lebih tinggi (6,81 ± 0,35 log sel/mL) daripada isolat bakteriofage SL3-St (7,39 ± 0,31 log sel/mL) dan S2-St (7,60 ± 0,27 log sel/mL). Penurunan densitas sel inang terendah oleh ketiga isolat bakteriofage terjadi pada jam ke-4. Bakteriofage B2-St merupakan bakteriofage terbaik dan berpotensi sebagai agen biokontrol Salmonella  Typhi. 
The effect of Phyllanthus niruri and Catharanthus roseus on Macrophage Polarization in Breast Cancer Mice Model: The Effect of P. niruri and C. roseus in Breast Cancer Mice Model Sakti, Sefihara Paramitha; Sari, Fikriya Novita; Rachmawati, Farida; Widyarti, Sri; Rahayu, Sri; Soewondo, Aris; Jatmiko, Yoga Dwi; Rifa'i, Muhaimin
Journal of Tropical Life Science Vol. 14 No. 1 (2024)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.14.01.03

Abstract

Cancer death cases have increased yearly, and there are estimated to be 21.6 million cancer cases in 2030. Studies of herbal compounds for cancer treatment alternatives are essential because cancer treatment is relatively expensive and has adverse effects. Phyllanthus niruri (Pn) and Catharanthus roseus (Cr) are plants that are known as herbal medicines. Combining the two plants is expected to prevent and enhance the immune system in breast cancer cases. This study aims to analyze the anti-cancer and immunomodulatory effects of P. niruri and C. roseus extract (PCE) in modulating macrophage polarization in breast cancer mice. Experimental animals are divided into six groups and there is healthy control (normal mice), cancer (DMBA-induced mice), cancer mice with cisplatin administration, cancer mice with PCE administration with three different doses, including dose 1 (500 mg/kg Pn + 15 mg/kg Cr), dose 2 (1000 mg/kg Pn + 75 mg/kg Cr), and dose 3 (2000 mg/kg Pn + 375 mg/kg Cr). The mice were injected with DMBA once a week for six weeks to induce cancer in mice. The breast cancer mice model was administered with PCE orally for 14 days. The expression of CD11b+IL-10+ and CD11b+IFN-γ+ demonstrated macrophage polarization. The results showed that breast cancer induction using DMBA increased the level of IL-10 and decreased the level of IFN-γ significantly compared to the normal group (p < 0.05). In specific doses, administration of PCE could reduce IL-10 levels and increase the level of IFN-γ significantly (p < 0.05). PCE can modulate the polarization of macrophages by suppressing the M2-like macrophage and increasing the M1-like macrophage. The ability of PCE to modulate macrophage polarization indicates that the combination of P. niruri and C. roseus has activity as an anti-cancer.
Microbial Succession and Sensory Enhancement in Robusta Wine Coffee Fermentation Jatmiko, Yoga Dwi; Mulasari, Ratna Agista Putri; Mustamin, Aryan; Puja, Lintang Ratu; Kanita, Anggita Bella Siez
Journal of Coffee and Sustainability Vol. 2 No. 1 (2025)
Publisher : Directorate of Research and Community Services

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jcs.2025.02.01.02

Abstract

Robusta wine coffee is a fermented product processed through multi-stage anaerobic fermentation to enhance its sensory quality through microbial activity. This study aimed to assess bacterial diversity and succession during fermentation, examine correlations between microbial populations and physicochemical parameters, and evaluate sensory attributes through organoleptic testing. Fermentation was carried out in five stages over 49 days, with bacterial isolation performed on PCA and MRS agar. Isolates were characterized morphologically, followed by Gram staining and API 50 CHL biochemical testing. Physicochemical parameters—pH, temperature, titratable acidity, moisture, and ash—were analyzed at each stage. A total of 26 isolates were recovered, with Simpson diversity index values <1, indicating dominance by certain isolates: A2, A8, A9, B4, D3, D8, and E3. Correlation analysis revealed positive relationships between bacterial counts and both moisture and pH, and negative correlations with temperature, ash, and titratable acidity. Biochemical profiling confirmed several dominant isolates as lactic acid bacteria (LAB): A9 as Lactobacillus fructivorans, B4 as L. delbrueckii, D3 as L. pentosus, D8 as Leuconostoc mesenteroides, and E3 as Lactococcus lactis. Cluster analysis based on phenotypic traits indicated strong similarity among LAB isolates, especially between D3 (L. pentosus) and the reference strain L. plantarum. Organoleptic testing demonstrated that wine coffee was preferred over non-fermented robusta, particularly in terms of aroma, acidity, and aftertaste. These results support the potential use of selected LAB isolates as starter cultures to enhance the sensory consistency and quality of wine coffee products.
PENINGKATAN KEAMANAN PANGAN DAN KUALITAS ORGANOLEPTIK IKAN ASAP KHAS DESA KARANGSARI TUBAN MELALUI INDUKSI PENGEMAS VAKUM Nurdiani, Rahmi; Jaziri, Abdul Aziz; Jatmiko, Yoga Dwi
JFMR (Journal of Fisheries and Marine Research) Vol. 4 No. 1 (2020): JFMR
Publisher : Faculty of Fisheries and Marine Science, Brawijaya University, Malang, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jfmr.2020.004.01.5

Abstract

Produksi ikan asap di Kabupaten Tuban tumbuh secara signifikan karena lokasi penjualannya dekat lokasi wisata.Salah satu kawasan penghasil ikan asap ini beradadi Desa Karangsari, Tuban. Permasalahan keamanan pangan masih belum diperhatikan oleh para produsen ikan asap yang terlihat dari tampilan produk dalam keranjang secara terbuka dan pengemasan yang seadanya.Hal ini memengaruhi daya simpan produk yang hanya bertahan 1-2 hari. Oleh karena itu, kegiatan Doktor Mengabdi Universitas Brawijayabertujuan untuk meningkatkan kualitas produk ikan asap (mikrobiologi dan organoleptik) melalui induksi teknologi pengemasan vakum pada kelompok usaha pengasapan ikan. Metode pelaksanaan program ini adalah penyediaan teknologi pengemasan vakum, penyuluhan dan pelatihan pada mitra, serta pendampingan dan pengawasan. Adanya peningkatan status keamanan pangan produk ikan asap melalui penerapan teknologi pengemasan vakum yang disimpan dalam tiga kondisi (suhu ruang, dingin dan beku) ini dievaluasi melalui uji kualitasbaik secara mikrobiologimaupun organoleptik. Hasil uji total bakteri dan organoleptik menunjukkan pengemasan vakum dapat mempertahankan kualitas produk menjadi lebih baik dan lebih diterima konsumen. Kombinasi antara pengemasan vakum dan penyimpanan pada suhu dingin danbeku dapat menghambat pertumbuhan bakteri secara optimal. Teknologi pengemasan vakum untuk produk ikan asap dapat meningkatkan keamanan pangan dannilai jual.
Pengembangan Protokol Deteksi Staphylococcus aureus Berbasis Molekuler Koentjoro, Maharani Pertiwi; Alviani, Melinda Nuril; Jatmiko, Yoga Dwi; Habibah, Laila Nur; Al Fatih, Ahmad Nuril Fuad; Kartikaningsih, Hartati
Bioscientist : Jurnal Ilmiah Biologi Vol. 12 No. 1 (2024): June
Publisher : Department of Biology Education, FSTT, Mandalika University of Education, Indonesia.

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33394/bioscientist.v12i1.9494

Abstract

Staphylococcus aureus becomes a normal flora in humans, especially on the skin and in the nose. However, if it becomes excessive or if there are pathogenic variants, it can cause various health problems. The purpose of the study is to develop a molecular-based detection method for Staphylococcus aureus using the norA primer gene. The norA gene in Staphylococcus aureus is known to play a role in pathogenesis with its antibiotic resistance ability. This type of research is analytical observational with a cross-sectional method. The methods in the study include the isolation and identification of Staphylococcus aureus from healthcare facility equipment. Isolation and identification include bacterial isolation using Blood Agar Plate (BAP) media; isolate purification, Gram staining; biochemical tests using Mannitol Salt Agar (MSA) media, glucose tests, Voges Proskauer (VP) tests, catalase tests, and coagulase tests. Furthermore, S. aureus isolates were tested using a molecular-based method, namely Polymerase Chain Reaction (PCR). This method includes DNA isolation stages, qualitative testing with agarose gel electrophoresis, semi-quantitative testing with image J software, amplification with Real-Time Polymerase Chain Reaction (RT-PCR) using norA gene primers. The Mann-Whitney test results gave a value of p = 0.334 (p> 0.05) indicating the suitability between the culture method and the PCR method with the developed protocol in detecting Staphylococcus aureus. The developed method includes the use of base sequences in the norA gene primer, optimization of annealing and extension temperatures, and the concentration of DNA templates used.
Detection of Plantaricin-Encoding Gene and Its Partial Purification in Lactobacillus plantarum BP102 Suryani, Elsa Mega; Jatmiko, Yoga Dwi; Mustafa, Irfan
Jurnal Biodjati Vol 8 No 2 (2023): November
Publisher : UIN Sunan Gunung Djati Bandung

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15575/biodjati.v8i2.27851

Abstract

Lactobacillus plantarum BP102 isolated from garlic bulb tissue has probiotic properties, including producing bacteriocin called plantaricin. This study aimed to detect the gene encoding bacteriocin produced by Lactobacillus plantarum BP102, and to evaluate the bacteriocin activity at each stage of partial purification. After the end of the log phase of L. plantarum BP102 was determined, and the bacteriocin-encoding genes were checked by PCR technique. Partial purification of bacteriocin was elucidated including pH-neutralized cell-free-supernatant (CFS), precipitation using 80% of ammonium sulfate, and dialysis (cut-off 10 kDa), then the bacteriocin activity in every partial purification stage was evaluated. The molecular weight of plantaricin was estimated using SDS-PAGE analysis. Lactobacillus plantarum BP102 harbored the gene encoding plantaricin (pln) biosynthesis, namely plnEF and plnK genes. The activity of crude bacteriocin was inactivated by the presence of proteinase-K enzyme. The protein concentration was gradually decreased along with the purification process. The bacteriocin activity was demonstrated at each step of the purification process (CFS, precipitation, and dialysis) against Bacillus cereus by 9.23 ± 0.20 mm, 7.86 ± 0.15 mm, and 7.6 ± 0.10 mm, respectively; while, Escherichia coli by 10.3 ± 0.55 mm, 7.4 ± 0.1 mm, and 6.86 ± 0.45, respectively. The molecular weight of partially purified bacteriocin BP102 was found to be approximately 15.9 kDa. The overlaid part of the gel showed a slight inhibition against E. coli due to a low protein concentration. This bacteriocin purification process should be further optimized to improve the bacteriocin activity that could be useful for food preservation.
Antibacterial Activity of Pluchea indica Leaf Extract was Increased After Being Fermented with Saccharomyces cerevisiae and Added with its Cell-Free Supernatant Rismawati, Ria; Jatmiko, Yoga Dwi; Widyarti, Sri
Biotropika: Journal of Tropical Biology Vol. 10 No. 2 (2022)
Publisher : Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.biotropika.2022.010.02.04

Abstract

Multi Drug Resistance (MDR) is a global health problem that endangers public health and can lead to death. Therefore, novel antibacterial agents are required derived from medicinal plants, one of them is beluntas (Pluchea indica) which has high potential as an antibacterial. Fermentation or addition of cell-free supernatant of Saccharomyces cerevisiae is thought to increase the antibacterial content of an herb. This study aimed to evaluate the effect of fermentation and the addition of Cell-Free Supernatant (CFS) of S. cerevisiae to enhance antibacterial activity of P. indica leaf extract. Fresh and dried leaves of beluntas were used in this study. The dried leaves in the form of powder was boiled at 100oC for 45 minutes. Fresh leaves were homogenized by blending. The extract of P. indica was used for fermentation and addition of CFS of S. cerevisiae. CFS as much as 60 mL dan fermentation with cell density of 7.53x105 CFU/mL with 100 mL of P. indica leaf extract were centrifuged at 10,000 rpm for 20 minutes. The antibacterial test method used was the Kirby-Bauer method against Escherichia coli and Staphylococcus aureus. The results showed an increase in antibacterial activity as indicated by the increasing diameter of the inhibition zone either by fermentation or addition of CFS with inhibition zone diameter was 3.85 – 4.81 mm against E. coli and 4.63 – 5.12 mm against S. aureus. The fermented P. indica and addition of CFS was shown to be potentially developed as antibacterial agents. Keywords: Antibacterial, CFS, fermentation, Pluchea indica, Saccharomyces cerevisiae