Indonesian Journal of Biotechnology
Not a member yet
387 research outputs found
Sort by
A simple method of plant sectioning using the agarose embedding technique for screening intracellular green fluorescent protein
No full textIt is difficult to observe plant tissue sections transformed using the agroinfiltration method under a fluorescent microscope. This is due to the softness of the post‐transformation plant. This research was conducted to optimize the sectioning of tobacco stems transformed through the agarose embedding technique. Optimization was conducted at various agarose concentrations: 2%, 4%, and 6%, followed by five minutes of incubation at various temperatures: –80 °C, 4 °C, and 25 °C. The stems were then cut using a scalpel and examined under a fluorescence microscope. The results showed that the embedding method using 6% agarose was more effective at producing a tobacco stem section than 2% or 4% agarose. Meanwhile, incubation at 25 °C was better suited to the transformed tobacco stems than at 4 °C or –80 °C. Green Fluorescent Protein (GFP) could be determined under a fluorescent microscope when using the optimum method. Thus, the optimum method for creating sections of transformed tobacco stems by embedding was to use 6% agarose followed by incubation at 25 °C for 5 min. The optimum result can be applied to obtain a slight section of tobacco stem in order to observe a recombinant protein or other anatomical structures
Fermentation medium optimization of Streptomyces sp. as an antifungal agent against the Ganoderma boninensis pathogen in oil palm
No full textGanoderma boninensis is the most common fungus which attacks oil palm trees. However, a significant percentage of inhibition to the problem is found through the use of Streptomyces sp. The optimization of the Streptomyces sp. fermentation medium growth factors affects the secondary metabolites production. This study aimed to identify the best formulation of carbon and nitrogen sources and the optimum concentration of Streptomyces sp. fermentation medium for antifungal compound production. The results showed that the best sources of carbon and nitrogen were liquid glucose and monosodium glutamate in the inhibition zones of 16.7 mm and 6.3 mm, while the best concentration levels were 20 g/L and 14.19 g/L, respectively. The results of the first optimization showed an inhibition zone response and area (%) of the optimum high‐performance liquid chromatography (HPLC) chromatogram of 24.39 mm and 62.68 percent, respectively. Taking the suggestion of the first optimization, the second optimization produced 15.2 g/L and 8.3 g/L. The predicted value of the inhibition zone was 21.47 mm, and the area (%) of the HPLC chromatogram was 53.44 percent. The validation results showed an inhibition zone response of 22.01 mm and an HPLC chromatogram area (%) of 54.86 percent. The difference between the predicted and validation values was less than 5 percent; the validation value was thus in line with the value predicted by Design Expert 10.0.7. The chemical formula of the probable active compound is that of the cyclo(phenylalanyl‐prolyl) compound
Production, purification and characterization of chitinase from Micromonospora sp. AR17
No full textN‐acetylglucosamine (NAG) is the monomer product of chitin, which has been widely used as a bioactive com‐ pound in applications such as anti‐tumor, anti‐microbial, and antioxidant activities. In production, biological processes using enzymes are preferable to chemicals due to environmental issues. This study aims to determine the activity, purity level, and molecular weight of purified chitinase from Micromonospora sp. AR17 determines the concentration of NAG produced by purified chitinase that has been characterized. Chitinase was produced by fermentation in colloidal chitin broth at 40 °C, pH 7, for 7 days, while chitinase activity was checked every 24 h. The optimal fermentation time was used to produce chitinase for a further purification step. Enzyme purification was carried out by ultrafiltration, ammonium sulfate precipitation, ion exchange chromatography (Q Sepharose Fast Flow), and gel filtration (Sephacryl S‐300). The purified enzyme was then char‐ acterized for optimum time, pH, and temperature to produce NAG. The results suggested that the fourth day was the optimal time for chitinase production, with chitinase activity of 0.0040 U/mL and a NAG concentration of 7.62 µg/mL. The purifica‐ tion step successfully increased the purity by 6.82 times with chitinase‐specific activity at 1.4648 U/mg. Production of NAG with purified chitinase produced a NAG concentration of 32.472 µg/mL with an incubation time of 30 min at 40 °C and pH 7
Effect of fluidised bed drying on ginsenoside content in hairy root cultures of Panax ginseng C.A. Meyer
No full textKorean Ginseng (Panax ginseng C.A. Meyer) is a high‐value herb with many pharmacological benefits due to its primary active compound, ginsenosides. The most ginsenosides are known to be thermolabile and susceptible to degradation at high‐temperature processing. Our previous studies revealed that the optimum parameters related to the P. ginseng tissue culture protocol, particularly for hairy root propagation of Cultured Roots of Mountain Ginseng (CRMG)‐88, was using a lab‐scale bioreactor. The next stage involves screening for a suitable post‐harvest treatment, i.e., drying, will be production of the best quality ginsenoside content. This study therefore aimed to examine the ginsenoside content by using a fluidised bed dryer (FBD) on the ginseng roots. Our results showed that FBD produced a significantly higher of total ginsenoside content (5.386 ± 1.167%), compared to control (3.750 ± 0.641%). FBD‐dried CRMG‐88 also appeared lighter in colour and more voluminous with a Loss on Drying (LOD) of 6.448 ± 1.900%. This study concluded that fluidised bed drying is superior in retaining ginsenoside content and has the potential for large‐scale application
A response surface methodology for the use of MIL‐101 as a catalyst for the one‐step synthesis of lactide
No full textLactide is a vital monomer for producing high molecular weight polylactic acid (PLA) through ring‐opening polymerization. This study synthesized crude lactide from L‐lactic acid with MIL‐101 as the catalyst. MIL‐101 is a metal‐based catalyst with organic ligands (MOF) that was prepared by reacting Cr(NO3)3.9H2O with terephthalic acid (BDC). The formation of MIL‐101 was confirmed from Fourier‐transform infrared (FTIR) analysis. The role of MIL‐101 and the effect of temperature, time, and MIL‐101 loading, as well as their interactions in the conversion of lactic acid to crude lactide, were then investigated using the response surface method (RSM). Crude lactide was analyzed using 1H‐nuclear magnetic resonance (NMR) spectroscopy to confirm the presence of lactide. The RSM results indicated that the highest conversion of 22.84% can be obtained using a temperature of 175 °C, 1.5% w/w MIL‐101 loading, and a reaction time of 5 h. The RSM model showed that the interaction of MIL‐101 loading and reaction time strongly affected the conversion of lactic acid to lactide, with a P‐value of 0.0021 and an F‐value of 50.45. In contrast, the interaction of catalyst loading and temperature did not significantly affect the conversion of lactic acid to lactide, with a P‐value of 0.2565 and an F‐value of 1.75
sgRNA design and in vitro nucleolytic analysis of the Cas9‐RNP complex for transgene‐free genome editing of the eIF4E1 gene from Capsicum an‐ nuum L.
No full textChili (Capsicum annuum L.) is a highly valued vegetable, renowned for its unique taste and aroma. However, chili production faces challenges in meeting the high demand due to infections caused by pathogens such as ChiVMV (potyvirus). Previous studies have suggested that chili eIF4E1 plays a crucial role in potyvirus gene transcription. Therefore, this study explores the potential of CRISPR‐Cas9‐based genome editing to enhance chili resistance by introducing premature stop codons or truncated proteins. Two sgRNAs were designed, targeting the first and second intron of the eIF4E1 gene. The production of Cas9 protein was assessed with varying IPTG concentrations in Escherichia coli BL21(DE3), carrying 4xNLS‐pMJ915v2‐sfGFP plasmid with a TEV protease cut‐site at the N terminal. The findings indicate that the optimal IPTG concentration is 500 µM. Purification using an IMAC column confirmed the presence of Cas9 in the initial 2 mL of the eluted fractions, as indicated by numerous background proteins. Nevertheless, successful formation of Cas9‐RNP complexes was achieved for both sgRNAs. The nucleolytic activity of Tag‐Cas9 (carrying the MBP‐tag) and Cas9 was confirmed through in vitro endonuclease activity assays. The next step involved transfecting chili protoplasts with these RNP complexes to edit the chili eIF4E1 gene
Atrazine degradation by Bacillus safensis strain BUK_BCH_BTE6 isolated from agricultural land in northwestern Nigeria
No full textAtrazine herbicide is known to disrupt the endocrine system and is potentially carcinogenic. Its continual application leads to high residue levels in soil, causing water pollution, which when consumed is associated with devastating health effects. This research reported the isolation and characterization of a new bacterial strains from active agricultural soil with the potential to biodegrade atrazine as a sole carbon source. An enrichment method was utilized to isolate the bacteria (A1, A2, B1, B2, C1 and C2) on mineral salt media (MSM) following serial dilution. Six isolates were screened for their tolerance to various concentrations of atrazine (500 to 1500 mg∙L‐1), and only isolate B1 tolerated up to 1500 mg∙L‐1 atrazine. The isolate was identified molecularly as Bacillus safensis strain BUK_BCH_BTE6 based on 16S rRNA gene sequencing and molecular phylogenetic analysis. Characterization of the isolate based on the effects of temperature, pH, substrate concentration, incubation time, inoculum size, and heavy metals revealed optimum growth and atrazine degradation at 35 °C, a pH of 7.5, 400 mg∙L‐1, at 48 h, and inoculum size of 600 µL, respectively. The growth of the isolate was inhibited by 2 ppm Hg, Cd, Cr, Pb, Ar, and Ni, while Fe, Cu, and Zn stimulated it. GC‐MS analysis revealed a degradation efficiency of 88.85% within 120 h, while metabolites such as desethyldeisopropylatrazine, deisopropylatrazine, N‐ethylammelide, and cyanuric acid were also detected. This isolate is a highly atrazine‐tolerant and efficient atrazine degrader that could be employed for bioremediation of atrazine‐polluted sites
Antibacterial activity of bioactive compound produced by endophytic fungi isolated from Mangifera casturi Kosterm endemic plant from South Kalimantan, Indonesia
No full textThe endophytic fungi that live in endemic plants are a promising bio‐prospect as the producers of antibacterial compounds. This research is aimed to evaluate the endophytic fungi antibacterial compound from Mangifera casturi. The bioactive compounds of 13 endophytic fungi were extracted using ethyl acetate and evaluated for antibacterial activity using disk diffusion assay. The minimum inhibitory concentration (MIC) was measured using the serial broth dilution method. Scanning Electron Microscopy (SEM) was used to examine cell damage because of the extract. The antibacterial compounds were then detected using GC‐MS analysis. The endophytic fungi were identified morphologically and molecularly based on ITS rDNA sequencing Among the 13 isolates, the endophytic fungi identified as Botryosphaeria rhodina AK32 produced the antibacterial compounds that exhibited the highest activity and a broad spectrum. Moreover, they were capable against resistant bacteria, Methicillin Resistant Staphylococcus aureus (MRSA) with an MIC value of 1.56% for all the test bacteria. The mechanism of action of AK32 ethyl acetate extract seemed to affect the condition of bacterial cell walls, causing morphological alteration such as shrinkage of the cell, warted cells, and hollow cells. Based on GC‐MS, the antibacterial compounds of AK32 ethyl acetate extract were di‐n‐octyl phthalate, benzyl alcohol, high‐oleic (CAS) safflower oil, benzene acetonitrile, and benzotriazole
Whole genome sequence analyses of Indonesian isolates SARS‐CoV‐2 variants and their clinical manifestations
No full textThe SARS‐CoV‐2 virus has been the cause of the global pandemic since the end of 2019. Since then, the virus has mutated to create different types of variants with numerous effects on those infected. This has complicated human intervention for prevention. Indonesia was heavily affected by the pandemic, specifically from May to August 2021, and as a country has recorded many distinct isolates. Thus, characterization of the virus strains from Indonesia is important. GISAID, NCBI BLAST, and MAFFT version 7 were used. There were 9,488 isolates in Indonesia as of November 2021, with the majority including the Delta variant. While most of the isolates have mutations common to those from other countries, there are some atypical ones, such as mutation V1264L in the Delta variant that was suspected to play a role in worsening the pandemic. The Delta variant had the most mutations in the spike protein when compared to the Alpha and Beta variants, giving it important roles in infectivity and vigorous entry into cells, with some general clinical manifestations like fever and sore throat; however, the severity of the Delta variant is attributable to its rapid growth. This is why, from May to November 2021 in Indonesia, cases of the Delta variant rocketed, unlike the other variants
Chitosan Xylotrupes gideon encapsulated lemongrass leaf ethanol extract reduce H2O2‐induced oxidative stress in human dermal fibroblast
No full textDuring phagocytosis, phagocyte cells discharge reactive oxygen species referred to as respiratory bursts, inducing a rise in pro‐oxidants and subjecting the cell to oxidative stress. Such stress is a biological mechanism related to an imbalance in pro‐oxidant/antioxidant homeostasis, which generates toxic reactive oxygen. Encapsulation is a coating process to improve the stability of bioactive compounds from lemongrass extract. Therefore, this study aims to determine the encapsulation activity of lemongrass leaf extract with chitosan X. gideon (LEChXg) to reduce the oxidative stress of fibroblasts. The research used the human dermal fibroblast (HDF) cell line, comprising negative and positive controls and use of LEChXg 100, 200, 300, 400, and 500 µg/mL. HDF cell migration was evaluated by employing the scratch wound healing method and the wound closure was oberseved at 0, 2, 4, 6, and 24 h intervals. The cell proliferation was observed at 24, 48, and 72 h using CCK‐8 at a 450 nm wavelength. The results showed that the observations at 0, 2, and 4 h did not demonstrate any significant difference on the cell migration (p > 0.05) among the groups. However, the wound closure at 4 and 6 h showed a significant difference (p < 0.05) with LEChXg 300 µg/mL. Despite the lack of any significant variation observed up to 24 h, fibroblast subjected to the stressor did not achieve complete closure. The groups treated with LEChXg were more stable in maintaining fibroblast proliferation up to the end of the observation than those with stressors at 24, 48, and 72 h. Fibroblast induced with a stressor was also more stable in maintaining migration and proliferation in groups receiving LEChXg 300 µg/mL