Indonesian Journal of Biotechnology
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The effect of ultrasonic processing on physical and chemical properties of milk‐based soft, brine cheese
No full textMany earlier studies have documented pasteurization problems in the dairy industry. As a result, ultrasonic processing has been researched as a non‐heat alternative to pasteurization. In this study, milk‐based soft cheese was treated using various sonication times (0, 1, and 3 min) at a set frequency (22 kHz) with an amplitude of 60% of 630 W and different ripening periods (0, 15, 30, and 60 days) in brine (15%), stored at 4 °C, to reduce heat treatment and increase yield. The physicochemical parameters of white cheeses were examined over next 60 days and compared with a control cheese. The result showed that ultrasound had no significant effect on the cheeses in terms of their fat and protein content on storage. Compared to the control sample, ultrasound treatment improved the taste and aroma ratings due to increased lipolysis and proteolysis. In terms of overall acceptability, the ultra‐filtrate cheese sonicated for 3 min received the highest marks compared to the control. Sonication for 3 min treated fresh milk showed the maximum yield (190.5 g/L milk) compared to untreated raw milk yields (150.32 g/L)
Hypoxic mesenchymal stem cell‐conditioned medium accelerates wound healing by regulating IL‐10 and TGF‐β levels in a full‐thickness‐wound rat model
No full textFull‐thickness wound healing is a complex process requiring a well‐orchestrated mechanism of various factors, including cytokines, particularly interleukin (IL)‐10 and transforming growth factor (TGF)‐β. IL‐10 and TGF‐β act as robust anti‐inflammatory cytokines in accelerating the wound healing process by regulating myofibroblasts. Hypoxic mesenchymal stem cell‐conditioned medium (hypMSC‐CM) containing cytokines potentially contribute to accelerate wound repair without scarring through the paracrine mechanism. This study aims to observe the role of hypMSC‐CM in controlling TGF‐β and IL‐10 levels to accelerate full‐thickness wound repair and regeneration. A total of 24 male Wistar rats were used in this study. Six healthy rats as a sham group and 18 rats were created as full‐thickness‐wound animal models using a 6 mm punch biopsy. The animals were randomly assigned into three groups (n = 6) consisting of two treatment groups treated with hypMSC‐CM at a low dose (200 µL hypMSC‐CM with 2 g water‐based gel added) and a high dose (400 µL hypMSC‐CM with 2 g water‐based gel added) and a control group (2 g water‐based gel only). The IL‐10 and TGF‐β levels were examined by ELISA. The results showed a significant increase in IL‐10 levels on day 3 after hypMSC‐CM treatment, followed by a decrease in platelet‐derived growth factor (PDGF) levels on days 6 and 9. In line with this finding, the TGF‐β levels also increased significantly on day 3 and then linearly decreased on days 6 and 9. HypMSC‐CM administra‐ tion may thus promote wound healing acceleration by controlling IL‐10 and TGF‐β levels in a full‐thickness‐wound rat model
Potential of marine sponge Jaspis sp.‐associated bacteria as an antimicrobial producer in Enggano Island
No full textSponges, a group of marine multicellular animals with a porous body structure, show potential for the production of bioactive compounds. Sponge‐associated bacteria are an alternative antimicrobial producer due to their high content of bioactive compounds. This study aimed to identify the highest‐potential antimicrobial‐producing bacteria isolate associated with Jaspis sp. sponges from Enggano island. The isolated bacteria were screened for antimicrobial activity against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans using cultures, supernatants, pellets, and crude extracts. The study also conducted genetic identification to determine the identity of the isolate with the greatest potency and its closest relationship using the 16S rRNA gene. The antimicrobial activity was determined by monitoring and measuring the diameter of the formed clear zones. The results of the observations of morphological characteristics revealed nine isolates from Jaspis sp. that each consisting of 6 JABS isolates and 3 JABB isolates. Based on isolates that had antimicrobial activity, JABS6 isolates had the best antimicrobial activity, with the diameter of inhibition zones of 24.7, 8.2, 4.6, and 33.7 mm for E. coli, P. aeruginosa, S. aureus and C. albicans, respectively. The genome sequencing of the JABS6 isolate confirmed that it was identical to Bacillus thuringiensis strain USS‐CAP‐1. The study concludes that this finding shows promise for the further development of future antimicrobial agents
The prevalence of KRAS and BRAF mutation in colorectal cancer patients in Bali
No full textMutations in the KRAS (Kirsten rat sarcoma viral oncogene homolog gene) and BRAF (v‐Raf murine sarcoma viral oncogene homolog B1) gene play a significant role in primary resistance to colorectal cancer therapy. Around 85‐90% of KRAS mutations in colorectal cancer occur in exon 2 (codon 12 and 13), whereas approximately 96% of BRAF mutations occur in exon 15 codon 600 (V600E). This study aimed to determine the prevalence and mutation characteristics of the KRAS and BRAF genes in colorectal cancer patients in Bali. The DNA was isolated from 44 formalin‐fixed paraffin‐embedded colorectal cancer samples which were stored in the Department of Pathology, Sanglah General Hospital in 2017. Detection of mutation was carried out by polymerase chain reaction (PCR) and direct sequencing. Out of 44 samples, only 27 were successfully amplified and sequenced. Our findings showed six samples (22.2%) with mutated KRAS at codons 12 and 13 (including two samples with G12D, one sample with G12V, and three samples with G13D). Interestingly, we found three samples (11.1%) of BRAF mutation, including two samples with V600E mutation and one with V600L mutation. Taken together, our results showed that KRAS and BRAF mutations were identified and occurred exclusively. Further studies are essential to identify the correlation of these mutations with colorectal cancer prognosis and response to chemotherap
Resistance gene expression in selected Indonesian pigmented rice varieties against infection by Xanthomonas oryzae pv. oryzae
No full textRice (Oryza sativa L.) production is limited by bacterial leaf blight (BLB), caused by Xanthomonas oryzae pv. oryzae (Xoo). For decades, researchers have attempted to control this disease by growing plants with blight‐resistant Xa genes. Genetic resources often vary between rice varieties, and there is little information about the genetic resources of the pigmented rice varieties widely grown in Indonesia and their resistance genes against Xoo. The purpose of this study was to determine the expression of Xa genes in pigmented rice such as Inpari 24 and Cempo Merah (red‐pigmented) along with Hitam Bantul (black‐pigmented) and white rice varieties IR64 and Ciherang, and to evaluate their resistance to BLB. All varieties carried the Xa4, Xa10 and xa13 genes but varied in the Xa1, Xa7 and Xa21 genes. The rice varieties expressed some of these genes only after inoculation with Xoo. Disease assessment categorised the three different pigmented rice varieties as resistant (Ciherang, Cempo Merah and Hitam Bantul), while IR64 (white) and Inpari 24 (red) were moderately resistant. There was no specific pattern of Xa genes possession, quality of expression or resistance level to X. oryzae pv. oryzae. Therefore, when breeding plants, the selection of parental variety must be considered in terms of the possession and expression of Xa genes such as Xa10 as a molecular marker for resistance
Purification and characterization of thermostable serine alkaline protease from Geobacillus sp. DS3 isolated from Sikidang crater, Dieng plateau, Central Java, Indonesia
No full textThermostable proteases that optimally withstand the high‐temperature conditions of thermophilic bacteria could be produced and purified, which would be highly beneficial for use in industry. Geobacillus sp. is a thermophilic bacterium that can be found in various environmental conditions. The goal of this study was to isolate and characterize thermostable serine protease that had been produced by thermophilic Geobacillus sp. strain DS3. The proteolytic index was measured in a solid medium. The expression of protease was optimized by Geobacillus sp. DS3 at 50 °C for 18 h. Targeted protease was purified using ammonium sulfate (40‐60%) and DEAE Sephadex A‐25 resin. Using SDS‐PAGE, the molecular weight of the enzyme was predicted to be around 32 kDa. Purified thermostable protease was highly activated at 70 °C, pH 9.6 stable for 1 h, and inhibited by PMSF. Therefore, this enzyme is classified as a thermostable alkaline serine protease. Its kinetic study revealed specific activity of 0.41 U/mg (Vmax) and 0.25 mg/mL (KM). Overall, a thermostable alkaline serine protease from Geobacillus sp. DS3 showed high activity at high temperatures and alkaline pH, which is vital for application in industries such as leather processing and detergent formulation
The design of Indonesian SARS‐CoV‐2 primers based on phylogenomic analysis of the SARS‐CoV‐2 clades
No full textMolecular detection needs to be augmented for COVID‐19 detection in Indonesia using the PCR method with primer‐based gene analysis. This is necessary because the RNA of the SARS‐CoV‐2 virus, the causative infectious agent of the pandemic, has been mutated. Therefore, this study aimed to develop a primer design for determining SARS‐CoV‐2 clades in Indonesia using phylogenomic analysis. Data were obtained from 38 GISAID (Global Initiative on Sharing All Influenza Data) viruses and the relationships were analyzed using maximum likelihood (ML) phylogenomic analysis with a substitution model of generalized time‐reversible (GTR) to construct the tree topology. The results showed that the five types of SARS‐CoVs‐2 clades in Indonesia were L, G, GH, GR, and O. It also indicated that the GH region had the highest rate of clade at 50%, with the S clade affecting its formation. Furthermore, the genome sequences of the GH type used to design its primer were based on three genes, namely RdRp, S, and N. The RdRp and N genes were found to be conserved and hardy mutants, while the S gene occurred repeatedly. Several previous studies have stated that the designed primers produced missense mutations compared to another in silico. Therefore, three sets of primers were achieved from the GC contents and clamps, Tm range, and structural secondary indicator standards
Expression of cytokinin responsive and ethylene biosynthesis genes in rice callus with different regeneration rates
No full textThis study aimed to investigate the correlation between callus regeneration rate and the expression of several genes responsible for cytokinin response and ethylene biosynthesis in the Ciherang, Mentik Wangi Susu, Hwayoung and Tarabas rice varieties. The callus regeneration rate of each rice variety was in vitro tested using N6 media, while the gene expression during the callus regeneration stages was examined using quantitative real‐time PCR (qRT‐PCR). Our results showed that the callus of Ciherang and Mentik Wangi Susu showed earlier green spot formation that then turned brown at a later stage, resulting in a low regeneration rate. While Hwayoung and Tarabas showed late green spot formation, high shoot regeneration was observed in both calluses. Gene expression analysis of regeneration media‐grown calluses showed that two cytokinin‐responsive genes, OsRR2 and OsRR6, were highly expressed in the Ciherang and Hwayoung callus, respectively. We also observed that ethylene biosynthesis genes such as OsACS1 and OsACO1 were highly expressed in the Mentik Wangi Susu and Hwayoung callus, respectively. Moreover, the expression of OsBBM1 was high in Hwayoung and Tarabas. Thus, the positive correlation between the expression of cytokinin‐responsive and ethylene biosynthesis genes with somatic embryogenesis activity likely depends on the induction level of OsBBM1
New sources of papain: SEM and SDS‐PAGE analysis to determine the natural tenderizer from papaya latex and senesced leaves
No full textThis study aims to determine the effectiveness of papaya‐fruit latex and yellow‐senesced leaves as a natural and organic tenderizer. The fruit and leaves of the plant were ground to powder, while 0 g, 10 g, 15 g and 20 g variations were used to cover 50 g of meat for 4 h. Subsequently, the Bradford and Kjeldahl methods were used to determine the protein content, while the protein profile was analyzed using SDS‐PAGE and confirmed using a Scanning Electron Microscope (SEM). The results showed that the protein concentration in mutton after fruit latex treatment was 41%, which was higher than the concentration of beef at 29.86%. Furthermore, the beef lost protein bands and its molecular weight fell from 225 kDa to 86 KDa, while the mutton experienced a reduction from 100 kDa to 65 kDa, which was significantly smaller than for raw meat. A single protein band was also observed at 21.6 kDa in the sample, indicating the presence of papain enzyme protein. Meanwhile, the SEM results showed that collagen and myofibril in the muscles were damaged in the treated meats. Based on these results, treatment with papaya fruit latex and yellow papaya leaves increases the tenderness of meat
Establishment of transgenic potato cultivar IPB CP1 plants containing gene encoding for superoxide dismutase to increase the abiotic stress tolerance
No full textPotato ( Solanum tuberosum L.) cultivar IPB CP1 is suitable as a raw material for the potato chip industry. Potato plants are sensitive to various abiotic stresses such as drought, aluminium and salinity, which induce reactive oxygen species (ROS). ROS is very toxic to plant cells. Superoxide dismutase (SOD) is one of the enzymes that catalyse ROS to H2O2 and O2. This study aimed to establish transgenic potato cv. IPB CP1 plants containing the MmCuZn‐SOD gene that are tolerant to various abiotic stresses. Genetic transformation using internodes without buds as explants produced putative transgenic potato with a transformation efficiency of 51.25% and a regeneration efficiency of 38.87%. Integration analysis of the MmCuZn‐SOD transgene in putative transgenic plants by polymerase chain reaction (PCR) with a set of specific primers showed that eight plants contained the MmCuZn‐SOD gene under the control of the 35S CaMV promoter. In vitro salinity stress, aluminium stress, and drought stress assays showed that transgenic plants had a higher number of roots and total root length than non‐transgenic ones. These results indicate that transgenic potato cv. IPB CP1 plants are more tolerant to abiotic stresses than non‐transgenic ones