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Isolation and Detection of Bacteriocin-Like Inhibitory Substances-Producing Bacteria from Fermented Mare’s Milk Sumbawa Mulyawati, Alifia Issabella; Ardyati, Tri; Jatmiko, Yoga Dwi
Jurnal Biodjati Vol 4 No 1 (2019): May
Publisher : UIN Sunan Gunung Djati Bandung

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15575/biodjati.v4i1.4194

Abstract

  Bacteriocin-like inhibitory substances (BLIS) produced by bacteria is a promising future food preservative agent. This study aimed to obtain bacterial strains that can produce broad-spectrum antibacterial agents and identify the best BLIS producer species based on 16S rDNA sequences. The bacterial strains were isolated from fer-mented mare’s milk using MRS and M17 agar medium. The isolates then were initially screened based on its antibacterial activity of crude cells against Staphylococcus aureus ATCC 6538. The selected strains were cultured and harvested for its cell-free supernatant (CFS). The pH of CFS was adjusted to 6.5 then used for antibacterial activity as-says against ten pathogenic bacteria. Also, the proteinaceous nature of BLIS compound was confirmed by testing with proteinase K. The gDNA of selected isolates was extracted and the 16S rDNA was am-plified using the polymerase chain reaction method then sequenced. The 16S rDNA sequences of the selected strains were used to identify the species using BLAST nucleotides from NCBI then the phylogenetic trees were constructed. 32 isolates was obtained, but only three iso-lates (BC9, SB7, and DC12) were selected as a result of antibacterial screening for further assays. The neutralized-CFS (N-CFS) of these isolates exhibited broad-spectrum antibacterial activity. The N-CFS could be assumed as BLIS. The isolate of BC9 was identified as Ba-cillus amyloliquefaciens strain BC9 that has 99.99 % similarity with B. amyloliquefaciens KC-1, SB7 was Lactobacillus plantarum strain SB7 that has 99.99 % similarity with Lb. plantarum JMC 1149T, and DC12 was Lactobacillus rhamnosus strain DC12 that has 100 % sim-ilarity with Lb. rhamnosus DSM 20021T. Thus, the BLIS produced by those strains is potential for future food and beverages preservations. 
Tofu Whey-Based Media for Probiotic Lactiplantibacillus plantarum D4 as a Halal Starter Culture Maulidiyah, Hikmah; Mustafa, Irfan; Jatmiko, Yoga Dwi; Ibrahim, Salam A.
Indonesian Journal of Halal Research Vol. 6 No. 1 (2024): February
Publisher : UIN Sunan Gunung Djati Bandung

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15575/ijhar.v6i1.30588

Abstract

In recent years, the development of probiotic-based fermented products with halal status has been a concern. The use of growth medium has relied on de Man Rogosa and Sharpe (MRS) as a relatively expensive standard medium, and its halal status is still uncertain. Extensive research has been carried out to investigate the development of low-cost halal alternative media for the cultivation of probiotic lactic acid bacteria (LAB). This study aimed to develop a probiotic halal and low-cost culture medium using a tofu whey-based medium. This study used three tofu whey-based media - A (tofu whey 100%), B (tofu whey 94.5%, molasses 3%, skim milk 2.5%), C (tofu whey 92.5%, molasses 3%, cheese whey 2.5%, tomato extract 2%), and MRS broth as a standard medium. Bacterial populations, total sugars utilized, total lactic acids produced, low pH (2.0) tolerance, and high bile salt concentration (oxgall 1.5%) were assayed. The highest bacterial population after 48 h of incubation was shown by medium B compared to medium MRS (12.34 ± 0.87 and 11.48 ± 0.3 log CFU/mL). Total sugars utilized by 48.28 ± 2.89, 38.89 ± 6.94, 39.14 ± 4.24, and 76.00 ± 1.41 %; on the other hand, total lactic acids produced by 0.16 ± 0.12, 0.03 ± 0.04, 0.31 ± 0.03, and 2.25 ± 1.48 % in A, B, C, and MRS, respectively. Probiotic tolerance at low pH and the presence of bile salts of Lactiplantibacillus plantarum D4 consistently showed a high survival rate in medium B compared to MRS. Based on these results, the components and proportions used in medium B were suitable for the growth of L. plantarum D4 as a halal probiotic starter candidate.
Development of a Stable CHO Cell Line for High-Level Expression and Purification of Recombinant Darbepoetin Alfa Widekdo, Dwi purno; Widodo, Nashi; Rifa'i, Muhaimin; Jatmiko, Yoga Dwi
Journal of Tropical Life Science Vol. 15 No. 3
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.15.03.12

Abstract

Recombinant Darbepoetin alfa (DARB), an erythropoiesis-stimulating agent, is widely used in treating anemia linked to chronic kidney disease and chemotherapy. This study presents a systematic approach for developing a high-yield, stable Chinese hamster ovary (CHO) cell line capable of producing recombinant DARB. A codon-optimized gene construct under dual CMV/EF1α promoters and puromycin resistance selection was transfected into CHO cells. To isolate high-producing monoclonal populations, transfected cells were subjected to limiting dilution cloning in 96-well plates, allowing single-cell-derived colonies to expand and be individually assessed. Clone DARV IV Pool 2 Clone 2E6 demonstrated the highest and most consistent DARB expression, validated through Western blot using anti-human EPO antibodies. The culture supernatant underwent two-step clarification via centrifugation and tangential flow filtration (TFF), followed by purification using anion-exchange chromatography on a HiTrap Q HP column. Gradient elution enabled effective separation, with SDS-PAGE and Western blot confirming high purity and molecular integrity of the recombinant protein. Variability in clone expression highlighted the influence of genomic integration sites and potential epigenetic silencing, emphasizing the importance of screening and stability validation. This study demonstrates that rational vector design, antibiotic-based clone selection, and robust purification strategies can produce CHO-derived DARB suitable for large-scale production. The integrated workflow supports scalability, product consistency, and regulatory readiness for biosimilar therapeutic manufacturing
Multidrug-resistant Bacteria in Domestic Cats (Felis catus): A Global Health Threat Emerging from Pets Mumtaza, Furzania; Jatmiko, Yoga Dwi; Salsabila, Maudy Syifa; Faturohmah, Yorda Nur; Fauziah, Ima; Saputra, Sugiyono
The Journal of Experimental Life Science Vol. 16 No. 1 (2026)
Publisher : Graduate School, Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jels.2026.016.01.01

Abstract

Cats (Felis catus) act as potential reservoirs for pathogenic bacteria due to their close contact with humans. Despite growing concerns about antimicrobial resistance (AMR) and zoonotic transmission, data on the diversity of antimicrobial-resistant bacteria, especially Staphylococcus aureus, in cats in Indonesia are limited. This study aimed to identify indicator bacteria in domestic cats and characterize their antimicrobial resistance (AMR) profiles. Nine earlobe swab samples were collected from cats at the Splendid Animal Market and Jalan Simpang Madukoro, Malang, in January 2023. White-toyellow colonies with  hemolytic activity formed on blood agar. Six antibiotics are used to asses antimicrobial susceptibility. Based on the European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoints, all isolates (100%) were resistant to rifampicin; seven isolates (78%) to oxacillin and cefoxitin; six isolates (67%) to gentamicin; four isolates (44%) to ceftriaxone; and one isolate (11%) to ciprofloxacin. Seven isolates (P1D, P3, P5, P7, R3, R5, and R9) were  resistance to more than three antibiotic classes were classified as multidrug-resistant bacteria (MDR). Molecular identification using 16S rRNA gene sequencing and phylogenetic analysis revealed the isolates as Chryseobacterium sp. (P1D), Enterobacter hormaechei (P3), Acinetobacter sp. (P5), Enterobacter cloacae (P7), Exiguobacterium sp. (P9A), Staphylococcus sciuri (R1A), Enterococcus hirae (R3), Pantoea sp. (R5), and Stenotrophomonas maltophilia (R9). The detection of ceftriaxone resistance and the presence of MDR bacterial strains indicate that cats carry resistant bacteria that may impact public health.
Characterization of Probiotics Isolated from Intestine of Mackerel Fish (Rastrelliger sp.) from Lembata Regency of East Nusa Tenggara Daten, Helena; Ardyati, Tri; Jatmiko, Yoga Dwi
The Journal of Experimental Life Science Vol. 10 No. 2 (2020)
Publisher : Graduate School, Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jels.2020.010.02.04

Abstract

The research aimed to isolate, characterize, and analyze the ability of lactic acid bacteria (LAB) potential as probiotics to produce hydrolase enzyme. The LAB was isolated using MRS agar by the spread plate method. The LAB characterization includes antimicrobial activity, tolerance to low pH, bile salt, salinity, autoaggregation properties, and ability to produce hydrolytic enzymes. The isolate which has the highest ability to inhibit Aeromonas hydrophila is KBP 3.3, while the isolate which inhibits the highest Streptococcus agalactiae is KBP 1.1.1. The KBP 3.3 and KBP 1.1.1 were able to survive at pH 1 for 24 hours with a survival rate of 93.6% and 98.3%. The KBP 3.3 and KBP 1.1.1 are tolerant to 7.5% bile salt concentrations for 24 hours of 99.46% and 99.11%. The KBP 3.3 is tolerant to 0.5 % salinity for 24 hours with the highest survival rate of 113.38%, while KBP 1.1.1 is 94%. The KBP 3.3 and 1.1.1 have autoaggregation properties of 92.18% and 87.84%. The KBP 3.3 produced the highest lipase enzyme, while KBP 1.1.1 produced the protease enzyme.Keywords: hydrolytic enzyme, lactic acid bacteria, mackerel, probiotic
Antimicrobial Activity of Combination Bacteriocin and Asam Sunti Extract (Averrhoa bilimbi L. fermented) Against Multidrug Resistant Escherichia coli in Lettuces (Lactuca sativa) Kimbal, Angie Via Resty; Jatmiko, Yoga Dwi; Ardyati, Tri
The Journal of Experimental Life Science Vol. 11 No. 2 (2021)
Publisher : Graduate School, Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jels.2021.011.02.06

Abstract

The ready-to-eat vegetables are often associated with the presence of multidrug-resistant (MDR) bacteria. This study aimed to evaluate the potency of bacteriocin, Asam Sunti extract, and their combination against MDR E. coli in lettuce. Their antimicrobial activity was assessed using the disk diffusion method and bacterial enumeration after direct application in pre-inoculated lettuce with MDR E. coli. The bacteriocin was produced by Lactobacillus plantarum BP102 at optimum production time or during the stationary phase at 18 h. These bacteriocins were able to inhibit five MDR E. coli isolates, while Asam Sunti extract and the combination of bacteriocin and Asam Sunti extract were only able to inhibit three MDR E. coli (LL1.2, LL3.11, and LL3.12) and (LL1.2, LL1.3, and LL3.11), respectively. In direct application to pre-inoculated fresh lettuce, higher inhibition of MDR E. coli was observed after applying the combination of bacteriocin and Asam Sunti extract with a ratio of 1:1 and 1:2, compared to bacteriocin alone. However, the inhibitory activity of this combination treatment was not significantly different (p>0.05) with the Asam Sunti extract alone. The highest rate of decrease in total bacteria in lettuces was 97% occurred in isolate LL1.2 with bacteriocin treatment alone, and isolate LL3.11 with combination treatment of bacteriocin and Asam Sunti extract (1:2). While on MCA media, the best reduction rate of 94% occurred in isolate LL1.2 with treatment using bacteriocin only, Asam Sunti extract only, and their combination (1:2). The inhibition of MDR E. coli in fresh lettuces by bacteriocin, Asam Sunti extract, and their combination was strain-dependent which was indicated by various inhibition results in all treatments.Key words: Asam Sunti extract, Bacteriocin, multidrug resistant.