HAYATI Journal of Biosciences
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Overexpression of Heat Shock Cognate Protein 71 kDa and Pyruvate Dehydrogenase in the Brain Tissue at the Early Stage of High Fat Diet Consumption
Get PDFHigh-fat diet (HFD) increases the risk of obese, while obesity increases the prevalence of metabolic syndrome and non-communicable diseases. Therefore, it will be interesting to evaluate the changes in metabolic parameters and brain profile upon the early consumption of HFD. In this study, a total of 12 Sprague-Dawley male rats were divided into two groups (n = 6), each group was fed with normal diet and HFD (39% of total calories from fats), respectively, for 6 weeks consecutively. The body weight, blood glucose, cholesterol and triglyceride values were measured. Furthermore, the brain and visceral adipose tissues were harvested at the end of the experiment. Protein was extracted from the brain tissue, and the protein extracts were separated by using two-dimensional gel electrophoresis and analyzed by liquid chromatography tandem mass spectrometric analysis (LC/MS/MS). In terms of food calorie, the rats fed with HFD consumed more energy than the rats fed with normal diet. Nevertheless, the blood triglyceride and cholesterol, and the visceral adipose tissue of both the HFD and normal diet fed rats were indifferent. At the molecular level, overexpression of stress proteins, namely heat shock cognate protein 71 kDa (Hsc70) and pyruvate dehydrogenase were detected in brain tissue of HFD group. These results suggest that HFD intake causing significant change in brain proteins profile at the early phase of its consumption when no clear metabolic changes were observed. This showed that the brain was affected by HFD
Biocontrol Activity, Mode of Action, and Colonization of Aureobasidium pullulans Dmg 30 DEP on Controlling Early Blight Disease on Tomato Plant
Get PDFWe investigated the biocontrol activity of A. pullulans Dmg 30 DEP against Alternaria solani causal agent of early blight. Biocontrol activity was tested by the in vivo and ad planta. Biocontrol activity were tested by investigating the antibiosis capabilities with dual culture method, paper dish assay, two-compartment petri dish assay, and trapping and identification of volatile organic compounds (VOCs) with GC-MS. Lysis activity was examined by observing the clear zone formed by growing yeast on chitin agar and skim milk agar. The ability of hyperparasitism was assessed by the agar block method, and observed by light microscopy and scanning electron microscopy (SEM). The results showed that A. pullulans Dmg 30 DEP plays a role in the suppression of early blight disease at 106 cells/ml and 107 cells/ml yeast cell density. The mechanism involved in biocontrol activity is the production of VOCs, the production of chitinase and protease enzymes, the production of siderophore and hyperparasitism. The result shows that A. pullulans Dmg 30 DEP was colonizing the tomato leaves following the areole
Glycoengineering of Darbepoetin-α in CHO-DG44 Cells through Overexpression of α-2,3-sialyl-transferase and CMP-sialic Acid Transporter
Get PDFSialic acid plays a very important role in determining the circulation life span of glycoprotein in various organisms. Therefore, having a high content of sialic acid is needed by glycoprotein therapeutic agents to be able to function as desired. For example, Darbepoetin (DPO), the 5 N-linked erythropoietin showed higher bioavailability and efficacy compared to 3 N-linked erythropoietin. However, in the DPO production process, the molecular weight can vary and is highly dependent on the content of sialic acid and its production host. To improve the DPO sialic acid contents in our CHO-DG44 expressing DPO, we have engineered the cells through overexpression of α-2,3-sialyl-transferase (ST) and CMP-sialic acid transporter (CST). The DPO contained in the supernatant of the engineered cells was analyzed by Western blot and characterized by using PNGase-F or neuraminidase enzyme digestions. The results showed that, two clones, overexpressing ST or CST, were obtained. The clones showed higher molecular weight of DPO as compared to DPO expressed by the parental cells, yet retained the same protein backbone. The overexpression of these two genes does not affect cell growth. This suggests that may be these cells beneficial for therapeutic glycoproteins
The Structural Annotations of The Mir-122 Non-Coding RNA from The Tilapia Fish (Oreochromis niloticus)
Get PDFTilapia (Oreochromis niloticus) is an important fisheries commodity. Scientific efforts have been done to increase its quality. One of them is staging a premium diet such as a fat-enriched diet. The transcriptomics approach is able to provide the signatures of the diet outcomes by observing the micro(mi)RNA signature in transcriptional regulation. Hence, it was found that the availability of mir-122 is essential in the regulation of a high-fat diet in tilapia. However, this transcriptomics signature is lacking structural annotations and the complete interaction annotations with its silencing(si)RNA. RNAcentral website was navigated for the latest annotation of mir-122 from tilapia and other species as a comparison. MEGA X was employed to comprehend the miRNA evolutionary repertoire. The RNA secondary structure prediction tools from the Vienna RNA package and the RNA tertiary structure prediction tools from simRNA and modeRNA are secured with default parameters. The HNADOCK tools were leveraged to observe the interaction between mir-122 and its siRNA. The post-processing was conducted with the Chimera visualization tool. The secondary and tertiary structure of the mir-122 and its siRNA could be elucidated, docked, and visualized. In this end, further effort to develop a comprehensive molecular breeding tool could be secured with the structural annotation information
Volatile Bioactive Compounds from Lasiodiplodia pseudotheobromae IBRL OS-64, an Endophytic Fungus Residing in the Leaf of Ocimum sanctum
Get PDFEndophytic fungi are known as potential novel compound producers with promising antimicrobial activities. Hence, the present study aimed to investigate the possible bioactive compounds present in the ethyl acetate extract of Lasiodiplodia pseudotheobromae IBRL OS-64. The ethyl acetate extract exhibited significant antibacterial activity against both Gram-positive and Gram-negative bacteria in disc diffusion assay. Thin-layer chromatography (TLC) was performed with chloroform, acetone and ethyl acetate (1:2:1, respectively) used as a solvent system and nine spots with diverse polarities were obtained. The TLC chromatogram with the active spot was localized with bioautography assay and the finding revealed that the dark spot with an Rf value of 0.5882 showed good antibacterial activity against all test bacteria. The fraction F5 exhibited promising antibacterial activity upon partial purification of dark spot via column chromatography and the GC-MS analysis of fraction F5 resulted in the detection of a major compound, 2-Benzenedicarboxylic acid, mono (2-ethylhexyl) ester with 90% matching factor. Thus, this compound may greatly contribute to the antibacterial activity of the fraction and has the potential to be developed as an antibiotic. The findings indirectly indicate that fungal endophytes from the medicinal plant could be a potential candidate for bioactive compounds with pharmaceutical properties
Ectoine and Hydroxyectoine from Halophilic Bacteria Isolated from Traditional Solar Saltern at Pejarakan Village, Buleleng Regency, Bali
Get PDFThe increased use of ectoine and hydroxyectoine in cosmetics and pharmaceuticals has led to the increasing demand for these bioactive compounds. This study aims to investigate the production of ectoine and hydroxyectoine by halophilic bacteria isolated from traditional solar saltern at Pejarakan Village, Buleleng Regency, Province of Bali, Indonesia. The halophilic bacteria were grown from the brine and the sediment samples using Luria Bertani media. The production of ectoine and hydroxyectoine was determined by inoculating the bacteria in MM63 media containing 12% w/v NaCl. Ectoine and hydroxyectoine were then extracted from the bacterial cells using a mixture of methanol/chloroform/water (10/5/4). Finally, the ability of the bacteria to excrete ectoine and hydroxyectoine was investigated using the osmotic downshock technique. The results showed that a total of 88 halophilic bacteria have been isolated which showed varied salt tolerances from 0 to 27.5% w/v. Most of the bacteria (86 isolates) were able to grow in MM63 media, suggesting the potential of the bacteria producing ectoine and hydroxyectoine. Further investigation showed that 33 selected halophilic bacteria were able to produce ectoine and hydroxyectoine with levels of 9.1 to 301.8 mg/L and 3.9 to 122.1 mg/L, respectively. All the bacteria were also able to excrete ectoine and hydroxyectoine after the osmotic downshock process with levels of 9.8 to 277.5 mg/L and 14.3 to 122.7 mg/L, respectively. The best ectoine and hydroxyectoine-producing bacteria showed the closest homology relationship with Salinivibrio costicola and Salinivibrio kushneri. Our study has found several promising candidates for ectoine and hydroxyectoine producers
Effect of the Coconut Coir (Cocos nucifera) as a Growth Medium for Pleurotus ostreatus (Oyster Mushroom) on Mineral and Vitamin B Contents
Get PDFThe oyster mushroom (Pleurotus ostreatus) is the most cultivated mushroom which its growth performance and nutritional composition depend on substrate types. Thus, this study investigated the effect of coconut coir on minerals and vitamins B contents in the oyster mushroom cultivation medium. The composition of coconut coir and Sengon’s sawdust as growth medium were variated at ratios of 4:0 (F1), 3:1 (F2), 2:2 (F3), 1:3 (F4), 0:4 (F5) (w/w). Furthermore, the mineral and vitamin B contents of the harvested oyster mushrooms were analyzed by ICPMS and LCMS, respectively. The highest minerals contents of potassium 26,909 mg/kg, 1,136 mg/kg of phosphorus, 313 mg/kg of magnesium, 4,346 mg/kg of calcium, 15.4 mg/kg for zinc, 2.07 mg/kg of copper, 0.623 mg/kg of Selenium were identified at F1, whereas the highest manganese was at F2 (15.3 mg/kg). Potassium was the highest mineral content in oyster mushrooms in all growth media, and low toxic minerals content of cadmium was detected between 0.015-0.058 mg/kg in all variant substrates. Lead (0.525 mg/kg) and mercury (0.012 mg/kg) was only detected at F1, while arsenic was detected at F4 (0.002 mg/kg) and F5 (0.029 mg/kg). LCMS analysis showed that vitamin B1 (Thiamine), B3 (niacin), B6 (pyridoxine) and B12 (cyanocobalamin) were detected in all growth media, while vitamin B5 (pantothenic acid) was only detected at F5. These results indicated that the P. ostreatus cultivation on coconut coir become valuable nutritional resources to alleviate malnutrition and help dispose of coconut coir in an environment-friendly manner
Cloning and Co-Expression of Lipase and its Specific Foldase from Ralstonia pickettii BK6
Get PDFThis study aimed to obtain a functional lipase (LipRM) from Ralstonia pickettii BK6 through a co-expression involving its foldase. The ORF of the LipRM was 999 bp while the gene encoding of lipase-specific foldase (LifRM) was 1,030 bp. LipRM and LifRM genes were cloned into a plasmid and were successfully co-expressed in Escherichia coli strains to produce functional LipRM. Enzyme activity from partially purified enzymes showed quite surprising results, LipRM activity in E. coli BL21 (DE3) was 25.84 U/ml, while the other strains (DH5α, HB101, S17-1λpir) were 628.98 U/ml, 761 U/ml, and 1206.46 U/ml, respectively. The highest relative activity of LipRM was found at 50-55°C and pH 7-8 with pNP-laurate (C12) as the preferred substrate specificity. LipRM activity was enhanced sharply in the presence of 30% organic solvents (methanol and ethanol) but decreased by more than 50% in the presence of detergents. This study was the first to report heterologous expression of Ralstonia pickettii lipase employing its native foldase resulting in functional lipase from subfamily I.2
Increasing The Stocking Density of Grouper Nurseries for Aquabusiness Efficiency in Recirculating Aquaculture System (RAS) with Bioremediation
Get PDFWater quality improvement was carried out by means of recirculation and bioremediation systems in grouper nurseries with high stocking density, thereby increasing production performance. The goal of this study was to evaluate the effectiveness of aquabusiness by analyzing the rise in grouper nursery stocking density using RAS and bioremediation systems. The test parameters consist of production performance, blood glucose levels, physic-chemistry-biology of water quality, and business efficiency based on productivity and business analysis. Treatment of 1,000 fish/m3 increased survival, blood glucose levels, total bacteria, Vibrio spp., ammonia, nitrite, and all productivity parameters. But it lowers all growth parameters, pH, and dissolved oxygen values. The recirculation system with bioremediation at up to 1,000 fish/m3 of stocking boosted the profit by 2.64 times. and 14.97% more efficient than 500 fish/m3 which could not be matched by the high production performance
Metabolites Profiling of Penicillium citrinum Recovered from Endophytic of Ramie (Boehmeria nivea) as a Potential Biocontrol Against Pathogenic Fungi
Get PDFEndophytes are a potent source of bioactive compounds that mimic plant-based metabolites. Fungi Fusarium spp. and Clonostachys rosea have been identified as pathogenic microorganisms in ramie (Boehmeria nivea). The antifungal test was carried out using the diffusion method and the MIC50 and Minimum Fungicidal Concentration (MFC) values using the tested microbial pathogens were Fusarium solani isolate 3248941, F. solani isolate Colpat-359, F. oxysporum isolate N-61-2, dan Clonostachys rosea strain B3042. Identification of secondary metabolites of the extract was carried out using GC-MS. The chromatogram of GC-MS analysis of this ethyl acetate extract (EA) showed seven dominant chemical compounds with various biological activities. Ethyl asetate extract of P. citrinum showed inhibition zone ranged from 19.10–22.07 mm with strong-very strong category at 200 mg ml-1 concentration against the tested microbial pathogens. Results revealed that the significant MIC values were observed against F. solani isolate 3248941 and Clonostachys rosea strain B3042 by less than 6.3 mg ml-1 and against F. solani isolate Colpat-359 and F. oxysporum isolate N-61-2 with 12.5 mg ml-1 and 25 mg ml-1 respectively. Minimum Fungicidal concentrations (MFC) for EA of P. citrinum were also reported against F. solani isolate 3248941 and C. rosea strain B3042, by 12.5 mg ml-1, whilst Fusarium solani isolate Colpat-359 by 25 mg ml-1 and Fusarium oxysporum isolate N-61-2 ,by 50 mg ml-1 respectively. Results of this study showed that P. citrinum, endophytic fungi of ramie, could be a promising source of compounds for antifungal agents