Indonesian Journal of Biotechnology
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Evaluation of synthetic‐gene recombinant LipL32 antigen for IgM ELISA detection of Leptospira infection
No full textLeptospirosis presents with nonspecific clinical features and requires time‐consuming laboratory tests for gold standard diagnosis. This study aims to design and characterize the recombinant LipL32 from synthetic gene and assess its performance as an antigen for detecting leptospirosis. The antigen was developed by cloning the LipL32 gene conserved portion of Leptospira interrogans serovar Icterohaemorrhagiae strain Langkawi. The immunoinformatic was used to characterize the developed rLipL32. Western blot results using anti‐histidine revealed a band of rLipL32 protein at ~40 kDa. Subsequently, it was used to examine the IgM antibody on human sera by using ELISA. The IgM‐LipL32 ELISA was evaluated using 67‐positive and 25‐negative sera and compared with a commercial ELISA. With a cut‐off value of 0.8, it showed 85.7% sensitivity, 83.3% specificity, a 48% positive prediction value (PPV), and 97% negative prediction value (NPV), indicating modest performance compared to existing commercial kits. The rLipL32 is a potential antigen for detecting IgM using ELISA; however, for use in low incidence areas, a confirmation test is crucial
Optimized bioethanol production from banana stem waste via simultaneous saccharification and fermentation with Saccharomyces cerevisiae
No full textIndonesia, one of the world’s largest banana producers, generates significant quantities of banana stem waste, leading to environmental challenges. This study explores the potential of converting this lignocellulosic biomass into bioethanol using a combination of steam pretreatment and simultaneous saccharification and fermentation (SSF) with Saccharomyces cerevisiae. The SSF process integrates enzymatic hydrolysis and fermentation, streamlining bioethanol production. The research applied the Taguchi method with an L9(34) orthogonal array to optimize key parameters, including enzyme concentration, particle size, temperature, and pH. Optimal conditions—5% enzyme concentration (v/v), 60 mesh banana powder, 35 °C and pH 5.00—yielded a maximum ethanol concentration of 9 g/L. Enzyme concentration and particle size were identified as critical factors in enhancing bioethanol yield. This study highlights the potential of banana stem waste as a sustainable resource for bioethanol production, contributing to waste reduction and renewable energy development
Integrated omics for increasing plant production and health‐related nutrition under extreme conditions: The Indonesia perspective
No full textDespite Indonesia being a megadiverse country that provides germplasm for breeding to produce improved future varieties, significant threats are faced related to biodiversity extinction. Such threats, for example habitat degradation and climate change, which lead to extreme conditions, must be addressed as they have contributed to stresses at the molecular level and affect plant production and health‐related nutrition. Integrated omics approaches have been applied to address the problems, as well as to produce varieties with superior traits, which are critical factors in achieving improved plant production and better naturally derived human nutrition. The paper discusses the omics research agenda in Indonesia; Indonesian biodiversity of nutraceutical plants and how omics can increase its production. Besides, current progress of omics application in Indonesia, policies and regulations to enhance integrated omics research are elucidated. By applying these approaches in Indonesia, breeding for better traits to support human needs and improve health quality will be greatly accelerated in the future
Mutations in RNA‐dependent RNA polymerase could be major cause of high pandemic potential of SARS‐CoV‐2: An in‐silico study
No full textHuman coronaviruses (HCoVs) are responsible for mild common cold to severe pneumonia‐like symptoms in infected individuals. The first HCoV was HCoV‐229E, discovered in 1962 in the US, which causes moderate symptoms. Since then, HCoVs have evolved, leading to epidemics or the recent SARS‐CoV‐2 pandemic. The main objective of this study was to understand the modifications occurred and what led to the transition from mild to pandemic form. Of the viral proteins, the RNA‐dependent RNA polymerase (RdRp) plays a crucial role in viral evolution, mutation, pathogenesis and transmission; this protein was therefore analyzed using in silico tools. We observed that RdRp has shown many mutations during its transition from mild to severe forms in HCoVs, which may have affected its enzymatic activity. The RdRp of HoV‐229E and HCoV‐NL63 showed 171 mutations, while SARS‐CoV‐2 showed the presence of 312. SARS‐CoV‐2 also showed a reduction in hydrophobic amino acid compared to the other HCoVs, consequently contributing to faster replication. Although mutations in the RdRp subdomains were found, yet five conserved regions was also presence among all the seven HCoVs; the finger and thumb subdomains had one conserved region, while the palm subdomains had three. Therefore, it can be inferred that on one hand the mutations reported in RdRp appeared to be the major cause of increased virulence leading to sporadic disease outbreaks, while on the other hand the presence of five conserved regions might prove to be potential targets for the development of new antiviral drugs
Biodegradable plant pots made from dried banana pseudo‐stems enriched with a Bacillus sp.‐biochar composite as an eco‐friendly alternative to plastic pots
No full textAgricultural plastic waste is a major environmental pollutant due to its non‐biodegradable nature. This study discusses the production of biodegradable pots (bio‐pots) using a biochar composed of banana pseudo‐stems and Bacillus sp. The isolated Bacillus sp. produced indole‐3‐acetic acid (IAA), solubilized potassium and phosphate, and secreted siderophores immobilized in banana pseudo‐stem biochar. X‐ray diffraction analysis revealed CaCO3 and KCl as the major elements, aside from carbon, released to the soil. Bio‐pots were made from banana pseudo‐stem biochar mixed with a Bacillus sp.–biochar composite at various formulations: 0%, 1%, 3%, 5%, and 10%. Mechanical testing indicated that the porous structure of the biochar contributed to low pot density and tensile strength. Moreover, the air‐filled spaces within the biochar enhanced water absorption, correlating with the amount of biochar used. Marigolds were cultivated outdoors in the bio‐pots to assess growth and yield. Our findings showed that those grown in biopot‐4 (10%) displayed improved growth and yield compared to the control group (grown in the ground). After 10 weeks, the control plants became infected with fungi and aphids, whereas those grown in biopot‐4 remained unaffected. In summary, bio‐pots incorporating 10% Bacillus sp.–biochar are eco‐friendly, reducing the need for chemical fertilizers, fungicides, and insecticides, while contributing to environmental sustainability. Moreover, the combination of biochar and Bacillus sp. is more effective than an unmixed form, since Bacillus sp. can inhabit and propagate in biochar pores if the conditions are otherwise unsuitable for growth
Moringa oleifera leaf extract ameliorates collagen degradation via the inhibition of MMP‐3 expression in UVB‐induced rats
No full textProlonged exposure to high‐intensity UVB induces the formation of reactive oxygen species (ROS) in skin tissue, triggering an increase in matrix metalloproteinase‐3 (MMP‐3) enzyme production and leading to collagen degradation. Moringa oleifera (MO) contains bioactive compounds known for ROS‐scavenging and anti‐inflammatory properties. However, the precise molecular mechanism of action remains unclear, requiring the inhibition of MMP‐3 activation and regulation of collagen deposition. This study aims to elucidate the potential effect of MO leaf extract‐based gel in restoring collagen deposition by reducing MMP‐3 activation in UVB irradiate‐induced collagen loss in rats. This study employed a completely randomized design, comprising four groups: a healthy group without UVB radiation, a negative control group subjected to UVB radiation and receiving a placebo, and two treatment groups exposed to UVB radiation with 5% or 10% moringa leaf extract‐based gel (MO‐5% or MO‐10%), respectively. Results showed that MO‐5% and MO‐10% significantly reduced MMP‐3 gene expression and increased collagen density compared to the negative control group (p < 0.05). Moringa oleifera leaf extract ameliorates collagen degradation by inhibiting MMP‐3 expression in UVB‐induced rats, suggesting its potential as a pharmacological and cosmetic agent for UVB‐induced skin damage
The diversity of fungal associates of Dendrobium ovatum (L.) Kraenzl., an endemic orchid of the Western Ghats of India
No full textDendrobium ovatum is a tropical epiphytic orchid endemic to the Western Ghats of India and has been listed as a threatened species in recent research due to its declining populations and changes in flowering and fruit set patterns. This study aims to investigate the mycoflora associated with the roots, stems and leaves of D. ovatum. Both surface‐associated and endophytic fungal associates were isolated and identified using morphological and molecular methods. The study resulted in the isolation of 139 cultures, which were divided into 24 morphotypes, 99% of which belonged to Ascomycota. The most dominant members, Trichoderma harzianum and Colletotrichum gloeosporioides, were consistently observed across all the study sites. Tissue‐specific fungal diversity analysis revealed that each organ was dominated by a distinct fungal group, forming characteristic communities specific to each tissue. The roots of D. ovatum exhibited the highest species richness and diversity, compared to the stem and leaves. This research also represents the first documentation of fungal associates of the threatened orchid D. ovatum
Effect of medium supplementations and extraction conditions on cellulase production through solid state fermentation of oil palm empty fruit bunches
No full textIn this study, cellulase enzyme was produced through solid state fermentation (SSF), employing oil palm empty fruit bunches (EFB) as the primary substrate. Two key aspects were explored to enhance crude enzyme yield, which are medium supplementation effects during SSF and cellulase recovery. During medium supplementation, glucose and/or Tween 80 were added alongside EFB substrate and other nutrients. Enzyme yield was determined using a filter paper assay and expressed as enzyme activity. The initial addition of glucose during fermentation led to increased crude enzyme activity, as measured by the filter paper assay. The peak crude enzyme activity was observed with the addition of 3 mg of glucose, with higher amounts showing no further increase in activity. Conversely, the addition of Tween 80 did not yield any significant increase in crude enzyme activity across all concentrations tested. The extraction conditions were varied to study cellulase recovery, specifically by adjusting the solid‐to‐solvent ratio and the number of extraction stages. Higher enzyme activity was achieved with lower solid‐to‐liquid ratios, as the increased solvent volume facilitated greater enzyme extraction. However, increasing the number of extraction steps did not significantly affect the resulting cellulase activity. Overall, this research underscores the need for further process optimization for cellulase production via SSF, utilizing the widely available EFB in Indonesia
Antidiabetic activity of Averrhoa bilimbi fruit methanol extract through enhancement of GLUT4 protein expression in diabetes‐induced mice
No full textGLUT4, the glucose transporter responsive to insulin, is primarily found in muscle and adipose tissues. Diabetes can result from impaired insulin secretion and sensitivity. This study aims to evaluate the antidiabetic properties of Averrhoa bilimbi methanol extract by enhancing GLUT4 protein expression in mice with induced diabetes. Extraction was conducted via soxhletation using 96% methanol. Phytochemical analysis, employing qualitative tests and GC‐MS, was performed. Antioxidant activity (IC50) and toxicity (LD50) were analyzed using DPPH and OECD methods. This research followed an experimental post‐only control group design, with mice intraperitoneally injected with 150 mg/kg BW of alloxan monohydrate. A total of 24 mice were then divided into six groups: Normal, Negative Control, Positive Control (metformin), Low Dose (50 mg/kg BW), Medium Dose (250 mg/kg BW), and High Dose (300 mg/kg BW). Treatment lasted 21 days, with fasting blood glucose and body weight measurements taken every three days. On day 21, the liver and skeletal muscle were isolated, and blood was collected. Serum insulin and GLUT4 expression were assessed via ELISA and Western Blot, respectively. Phytochemical screening revealed flavonoids, saponins, terpenoids, tannins, and phenols and their derivatives. The IC50 value was 85 µg/mL, with an LD50 value of 1,000 mg/kg BW, indicating strong antioxidant activity and mild toxicity. The extract significantly reduced blood glucose levels but did not impact weight loss in diabetic mice. Average liver weight and index were highest in the Negative Control group, yet the lowest levels of hepatic and muscle glycogen were also observed in this group. Interestingly, insulin level and HOMA‐IR decreased in diabetic mice, while the Medium Dose group exhibited the highest GLUT4 expression levels. In conclusion, medium doses of A. bilimbi methanol extract hold potential for diabetes treatment, with a probable mechanism of targeting GLUT4 protein expression
Easy extraction of Ganoderma boninense liquid sample using portable on‐chip device
No full textDetecting Ganoderma boninense in Indonesia is crucial for effectively controlling and mitigating the spread of basal stem disease in oil palm fields. While there is ongoing development of tolerant plants, no such plant has been successfully created yet. Consequently, researchers are actively studying detection methods for Ganoderma boninense. One established and highly accurate approach is the use of polymerase chain reaction (PCR) techniques for molecular detection. However, this method requires time‐consuming sample preparation, which can pose challenges in plantation settings. To address this problem, a portable lab‐on‐chip device has been introduced. This technology enables easy and automatic DNA retrieval from liquid samples by absorbing lysed DNA using magnetic beads. An efficient mechanism for manipulating the magnetic bead within the semiconductor has been successfully implemented. The extraction process typically takes around 15 minutes using a modified methodology on the chip device approach. The chip facilitates the retrieval of two samples with a capacity of 120 µL for each sample. The PCR method was utilized to validate the equivalence of the lab‐on‐chip device extraction to the standard extraction method. This represents a promising alternative for expedited and simplified detection of Ganoderma boninense in field conditions