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Fulltext Partial purification and characterization of the molybdenum-reducing enzyme from the glyphosate-degrading Burkholderia vietnamiensis strain Aq5-12

Jamulidin, S.N.K., Manogaran. M., Yakasai, M.H., Rahman, M.F.A., Shukor, M.Y.

Bioremediation Science and Technology Research, 2016;4(1):7-9.
MyJurnal

In this study, a novel glyphosate-degrading shows the ability to reduce molybdenum to
molybdenum blue. The enzyme from this bacterium was partially purified and partially
characterized to ascertain whether the Mo-reducing enzyme from this bacterium shows better or
lower efficiency in reducing molybdenum compared to other Mo-reducing bacterium that only
exhibits a single biotransformation activity. The enzyme was partially purified using ammonium
sulphate fractionation. The Vmax for the electron donating substrate or NADH was at 1.905 nmole
Mo blue/min while the Km was 6.146 mM. The regression coefficient was 0.98. Comparative
assessment with the previously characterized Mo-reducing enzyme from various bacteria showed
that the Mo-reducing enzyme from Burkholderia vietnamiensis strain AQ5-12 showed a lower
enzyme activity.
Gynura procumbens ethanol extract and its fractions inhibit macrophage derived foam cell formation

Manogaran M, Vuanghao L, Mohamed R

J Ethnopharmacol, 2020 Mar 01;249:112410.
PMID: 31747560 DOI: 10.1016/j.jep.2019.112410

ETHNOPHARMACOLOGY RELEVANCE: Gynura procumbens (Lour.) Merr. displayed cardio-protective effect that may prevent atherogenesis. The primary underlying pathological process of cardiovascular disease is atherosclerosis. Atherosclerotic lesion composed of macrophages, T cells and other immune cells which incorporated with cholesterol that infiltrates from the blood.
AIM OF THE STUDY: The present study was performed to determine underlying mechanism of G. procumbens ethanol extract and its fractions such as aqueous, chloroform, ethyl acetate and hexane affect macrophage derived foam cell formation.
MATERIALS AND METHODS: Lipid droplets accumulation in treated macrophages were visualized by Oil Red O staining while the total cholesterol present in the treated macrophages were measured using Cholestryl Ester quantification assay kit. Enzyme-Linked Immunosorbent Assay (ELISA) were used to detect TNF-α and IL-1β secretion in the supernatant of treated macrophages. Gene expression of Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) and ATP-binding cassette transporter A-1 (ABCA-1) in treated macrophages were analyzed using Real-Time Quantitative Polymerase Chain Reaction (RT-qPCR).
RESULTS: G. procumbens ethanol extract and its fractions reduced lipid droplet accumulation and total cholesterol in oxLDL-treated macrophages together with significantly reduction of TNF-α and IL-1β secretions in supernatant oxLDL-treated macrophages. LOX-1 gene expression was significantly reduced when G. procumbens ethanol extract and its fractions were added in oxDL-treated macrophages. In contrast, G. procumbens ethanol extract and its fractions significantly increased the expression of ABCA-1 gene in oxLDL-treated macrophages.
CONCLUSION: In conclusion, G. procumbens ethanol extract and its fractions inhibit the formation of macrophage derived foam cell by reducing TNF-α and IL-1β expression, which usually highly expressed in atherosclerotic plaques, suppressing scavenger receptor LOX-1 gene that binds oxLDL but induced ABCA-1 gene that mediate lipid efflux from macrophages.
Fulltext Improvement of Ficin-Based Inhibitive Enzyme Assay for Toxic Metals Using Response Surface Methodology and Its Application for Near Real-Time Monitoring of Mercury in Marine Waters

Uba G, Manogaran M, Gunasekaran B, Halmi MIE, Shukor MYA

Int J Environ Res Public Health, 2020 11 19;17(22).
PMID: 33227985 DOI: 10.3390/ijerph17228585

Potentially toxic metals pollution in the Straits of Malacca warrants the development of rapid, simple and sensitive assays. Enzyme-based assays are excellent preliminary screening tools with near real-time potential. The heavy-metal assay based on the protease ficin was optimized for mercury detection using response surface methodology. The inhibitive assay is based on ficin action on the substrate casein and residual casein is determined using the Coomassie dye-binding assay. Toxic metals strongly inhibit this hydrolysis. A central composite design (CCD) was utilized to optimize the detection of toxic metals. The results show a marked improvement for the concentration causing 50% inhibition (IC50) for mercury, silver and copper. Compared to one-factor-at-a-time (OFAT) optimization, RSM gave an improvement of IC50 (mg/L) from 0.060 (95% CI, 0.030-0.080) to 0.017 (95% CI, 0.016-0.019), from 0.098 (95% CI, 0.077-0.127) to 0.028 (95% CI, 0.022-0.037) and from 0.040 (95% CI, 0.035-0.045) to 0.023 (95% CI, 0.020-0.027), for mercury, silver and copper, respectively. A near-real time monitoring of mercury concentration in the Straits of Malacca at one location in Port Klang was carried out over a 4 h interval for a total of 24 h and validated by instrumental analysis, with the result revealing an absence of mercury pollution in the sampling site.
Optimisation of culture composition for glyphosate degradation byBurkholderia vietnamiensisstrain AQ5-12

Manogaran M, Shukor MY, Yasid NA, Khalil KA, Ahmad SA

3 Biotech, 2018 Feb;8(2):108.
PMID: 29430369 DOI: 10.1007/s13205-018-1123-4

The herbicide glyphosate is often used to control weeds in agricultural lands. However, despite its ability to effectively kill weeds at low cost, health problems are still reported due to its toxicity level. The removal of glyphosate from the environment is usually done by microbiological process since chemical process of degradation is ineffective due to the presence of highly stable bonds. Therefore, finding glyphosate-degrading microorganisms in the soil of interest is crucial to remediate this glyphosate.Burkholderia vietnamiensisstrain AQ5-12 was found to have glyphosate-degrading ability. Optimisation of biodegradation condition was carried out utilising one factor at a time (OFAT) and response surface methodology (RSM). Five parameters including carbon and nitrogen source, pH, temperature and glyphosate concentration were optimised. Based on OFAT result, glyphosate degradation was observed to be optimum at fructose concentration of 6, 0.5 g/L ammonia sulphate, pH 6.5, temperature of 32 °C and glyphosate concentration at 100 ppm. Meanwhile, RSM resulted in a better degradation with 92.32% of 100 ppm glyphosate compared to OFAT. The bacterium was seen to tolerate up to 500 ppm glyphosate while increasing concentration results in reduced degradation and bacterial growth rate.
Characterisation of the simultaneous molybdenum reduction and glyphosate degradation byBurkholderia vietnamiensisAQ5-12 andBurkholderiasp. AQ5-13

Manogaran M, Ahmad SA, Yasid NA, Yakasai HM, Shukor MY

3 Biotech, 2018 Feb;8(2):117.
PMID: 29430378 DOI: 10.1007/s13205-018-1141-2

In this novel study, we report on the use of two molybdenum-reducing bacteria with the ability to utilise the herbicide glyphosate as the phosphorus source. The bacteria reduced sodium molybdate to molybdenum blue (Mo-blue), a colloidal and insoluble product, which is less toxic. The characterisation of the molybdenum-reducing bacteria was carried out using resting cells immersed in low-phosphate molybdenum media. Two glyphosate-degrading bacteria, namelyBurkholderia vietnamiensisAQ5-12 andBurkholderiasp. AQ5-13, were able to use glyphosate as a phosphorous source to support molybdenum reduction to Mo-blue. The bacteria optimally reduced molybdenum between the pHs of 6.25 and 8. The optimum concentrations of molybdate for strainBurkholderia vietnamiensis strainAQ5-12 was observed to be between 40 and 60 mM, while forBurkholderiasp. AQ5-13, the optimum molybdate concentration occurred between 40 and 50 mM. Furthermore, 5 mM of phosphate was seen as the optimum concentration supporting molybdenum reduction for both bacteria. The optimum temperature aiding Mo-blue formation ranged from 30 to 40 °C forBurkholderia vietnamiensis strainAQ5-12, whereas forBurkholderiasp. AQ5-13, the range was from 35 to 40 °C. Glucose was the best electron donor for supporting molybdate reduction, followed by sucrose, fructose and galactose for both strains. Ammonium sulphate was the best nitrogen source in supporting molybdenum reduction. Interestingly, increasing the glyphosate concentrations beyond 100 and 300 ppm forBurkholderia vietnamiensis strainAQ5-12 andBurkholderiasp. AQ5-13, respectively, significantly inhibited molybdenum reduction. The ability of these bacteria to reduce molybdenum while degrading glyphosate is a useful process for the bioremediation of both toxicants.
Fulltext Microbiological Reduction of Molybdenum to Molybdenum Blue as a Sustainable Remediation Tool for Molybdenum: A Comprehensive Review

Yakasai HM, Rahman MF, Manogaran M, Yasid NA, Syed MA, Shamaan NA, et al.

Int J Environ Res Public Health, 2021 May 27;18(11).
PMID: 34071757 DOI: 10.3390/ijerph18115731

Molybdenum (Mo) microbial bioreduction is a phenomenon that is beginning to be recognized globally as a tool for the remediation of molybdenum toxicity. Molybdenum toxicity continues to be demonstrated in many animal models of spermatogenesis and oogenesis, particularly those of ruminants. The phenomenon has been reported for more than 100 years without a clear understanding of the reduction mechanism, indicating a clear gap in the scientific knowledge. This knowledge is not just fundamentally important-it is specifically important in applications for bioremediation measures and the sustainable recovery of metal from industrial or mine effluent. To date, about 52 molybdenum-reducing bacteria have been isolated globally. An increasing number of reports have also been published regarding the assimilation of other xenobiotics. This phenomenon is likely to be observed in current and future events in which the remediation of xenobiotics requires microorganisms capable of degrading or transforming multi-xenobiotics. This review aimed to comprehensively catalogue all of the characterizations of molybdenum-reducing microorganisms to date and identify future opportunities and improvements.
Fulltext Co-anaerobic digestion of sawdust and chicken manure with plant herbs: Biogas generation and kinetic study

Hakimi M, Manogaran MD, Shamsuddin R, Mohd Johari SA, Abdalla M Hassan M, Soehartanto T

Heliyon, 2023 Jun;9(6):e17096.
PMID: 37342579 DOI: 10.1016/j.heliyon.2023.e17096

Plant herbs specifically serai wangi (SW) and peppermint (PPM) are selected for its insect repellent properties as the use of chicken manure (CM) in anaerobic digestion (AD) potentially attract flies due to the digestate produced. Hence, the addition of SW and PPM in the AD system of CM could deter flies' infestation while producing biogas. Previous work has shown that AD of sawdust (SD) and CM with these plant herbs were able to produce biogas and reduce the flies attraction towards the digestate. However, the combination of SW and PPM for AD of CM has yet to be investigated. This work describes the effect of mixing SW and PPM on the co-AD of SDCM with respect to biogas production, methane yield and kinetic analysis. The mixture of SW and PPM was varied at different concentrations. The composition of methane in biogas was characterized every 10 days by using gas chromatography (GC) equipped with a thermal conductivity detector (TCD). The results suggest that co-AD of 10SW10PPM exhibited the highest biogas production (52.28 mL/gvs) and methane yield (30.89 mL/gvs), which the purity of methane increased by 18.52% as compared to SDCM. However, increasing the concentration of SW and PPM does not significantly improve the overall process. High R2 (0.927-0.999), low RMSE (0.08-0.61) and low prediction error (<10.00%) were displayed by the modified Gompertz, logistic and Cone models. In contrast, Monod and Fitzhugh model is not preferred for the co-AD of SDCM with a mixture of SW and PM, as a high prediction error is obtained throughout the study. Increasing the dosage of PPM decreases the maximum cumulative methane yield, ranging from 31.76 to 7.01 mL/gvs for modified Gompertz and 89.56 to 19.31 mL/gvs for logistic model. The Modified Gompertz obtained a lag phase of 10.01-28.28 days while the logistic model obtained a lag phase of 37.29-52.48 days.
Fulltext Biodecolourisation of Reactive Red 120 as a Sole Carbon Source by a Bacterial Consortium-Toxicity Assessment and Statistical Optimisation

Manogaran M, Yasid NA, Othman AR, Gunasekaran B, Halmi MIE, Shukor MYA

Int J Environ Res Public Health, 2021 Mar 02;18(5).
PMID: 33801387 DOI: 10.3390/ijerph18052424

The application of microorganisms in azo dye remediation has gained significant attention, leading to various published studies reporting different methods for obtaining the best dye decolouriser. This paper investigates and compares the role of methods and media used in obtaining a bacterial consortium capable of decolourising azo dye as the sole carbon source, which is extremely rare to find. It was demonstrated that a prolonged acclimation under low substrate availability successfully isolated a novel consortium capable of utilising Reactive Red 120 dye as a sole carbon source in aerobic conditions. This consortium, known as JR3, consists of Pseudomonas aeruginosa strain MM01, Enterobacter sp. strain MM05 and Serratia marcescens strain MM06. Decolourised metabolites of consortium JR3 showed an improvement in mung bean's seed germination and shoot and root length. One-factor-at-time optimisation characterisation showed maximal of 82.9% decolourisation at 0.7 g/L ammonium sulphate, pH 8, 35 °C, and RR120 concentrations of 200 ppm. Decolourisation modelling utilising response surface methodology (RSM) successfully improved decolourisation even more. RSM resulted in maximal decolourisation of 92.79% using 0.645 g/L ammonium sulphate, pH 8.29, 34.5 °C and 200 ppm RR120.
Life cycle assessment: Sustainability of biodiesel production from black soldier fly larvae feeding on thermally pre-treated sewage sludge under a tropical country setting

Seng Liew C, Ren Mong G, Wei Lim J, Raksasat R, Rawindran H, Hong Leong W, et al.

Waste Manag, 2023 Apr 20;164:238-249.
PMID: 37086606 DOI: 10.1016/j.wasman.2023.04.013

More energy is needed nowadays due to global population growth. Concurrently, sewage sludge generation has also increased steadily stemming from the inevitable urbanization. As such, black soldier fly larvae (BSFL) can be potentially deployed to solve both issues. This paper investigates the environmental sustainability of biodiesel production derived from sludge-fed BSFL feedstock. A cradle-to-gate life cycle assessment (LCA) was performed through SimaPro software utilizing the ReCiPe 2016 Midpoint (H) and Endpoint (H) methods. The entire LCA covered 3 main stages, including the thermal pre-treatment of sludge, BSFL rearing and processing, and lastly lipid extraction and biodiesel production. LCA showed that the sludge pre-treatment stage had the highest environmental impact, while BSFL rearing and processing had the least due to the suitable geographical climate. Electricity usage during the pre-treatment stage was the main contributing component, followed by chemical usage during biodiesel production. After normalizing, it was observed that land occupation, marine ecotoxicity, freshwater ecotoxicity and freshwater eutrophication were more impactful than the commonly studied global warming potential (GWP). Lipid content and biodiesel conversion efficiency were determined as the sensitive factors which could influence the LCA outcome. In comparison with other types of biodiesel, BSFL biodiesel had a milder impact in terms of climate change, land occupation, terrestrial acidification, marine and freshwater eutrophication. Furthermore, this biological reduction of sludge through BSFL valorization avoided sludge landfilling, which reduced up to 100 times GWP. Therefore, sludge-fed BSFL biodiesel production is an environmentally-sound and highly potential solution that should be investigated comprehensively.