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  1. Fulltext Extraction, characterization and molecular structure of bovine skin gelatin extracted with plant enzymes bromelain and zingibain
    Ahmad T, Ismail A, Ahmad SA, Khalil KA, Kee LT, Awad EA, et al.
    J Food Sci Technol, 2020 Oct;57(10):3772-3781.
    PMID: 32903957 DOI: 10.1007/s13197-020-04409-2
    Bovine skin was incubated with plant enzymes bromelain (B) and zingibain (Z) at the level of 0, 5, 10, 15, 20 and 25 unit/g of skin and gelatin was extracted at 60 °C for 6 h. Control gelatin was extracted without enzymatic pretreatment. The yield and gel strength were 17.90% and 283.35 g for the control samples and 22.26% and 160.88 g for B20 samples. The zingibain extracted gelatin (GEZ) samples failed to form gel. Viscosities of GEZ gelatins were significantly (P 
  2. 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.
  3. Fulltext Evaluation of conventional and response surface level optimisation of n-dodecane (n-C12) mineralisation by psychrotolerant strains isolated from pristine soil at Southern Victoria Island, Antarctica
    Habib S, Ahmad SA, Johari WLW, Shukor MYA, Alias SA, Khalil KA, et al.
    Microb Cell Fact, 2018 Mar 17;17(1):44.
    PMID: 29549881 DOI: 10.1186/s12934-018-0889-8
    BACKGROUND: Biodegradation of hydrocarbons in Antarctic soil has been reported to be achieved through the utilisation of indigenous cold-adapted microorganisms. Although numerous bacteria isolated from hydrocarbon-contaminated sites in Antarctica were able to demonstrate promising outcomes in utilising hydrocarbon components as their energy source, reports on the utilisation of hydrocarbons by strains isolated from pristine Antarctic soil are scarce. In the present work, two psychrotolerant strains isolated from Antarctic pristine soil with the competency to utilise diesel fuel as the sole carbon source were identified and optimised through conventional and response surface method.

    RESULTS: Two potent hydrocarbon-degraders (ADL15 and ADL36) were identified via partial 16S rRNA gene sequence analysis, and revealed to be closely related to the genus Pseudomonas and Rhodococcus sp., respectively. Factors affecting diesel degradation such as temperature, hydrocarbon concentration, pH and salt tolerance were studied. Although strain ADL36 was able to withstand a higher concentration of diesel than strain ADL15, both strains showed similar optimal condition for the cell's growth at pH 7.0 and 1.0% (w/v) NaCl at the conventional 'one-factor-at-a-time' level. Both strains were observed to be psychrotrophs with optimal temperatures of 20 °C. Qualitative and quantitative analysis were performed with a gas chromatograph equipped with a flame ionisation detector to measure the reduction of n-alkane components in diesel. In the pre-screening medium, strain ADL36 showed 83.75% of n-dodecane mineralisation while the reduction of n-dodecane by strain ADL15 was merely at 22.39%. The optimised condition for n-dodecane mineralisation predicted through response surface methodology enhanced the reduction of n-dodecane to 99.89 and 38.32% for strain ADL36 and strain ADL15, respectively.

    CONCLUSIONS: Strain ADL36 proves to be a better candidate for bioaugmentation operations on sites contaminated with aliphatic hydrocarbons especially in the Antarctic and other cold regions. The results obtained throughout strongly supports the use of RSM for medium optimisation.

  4. Fulltext Facile Coaxial Electrospinning Synthesis of Polyacrylonitrile/Cellulose Acetate Nanofiber Membrane for Oil-Water Separations
    Al-Rajabi MM, Almanassra IW, Khalil AKA, Atieh MA, Laoui T, Khalil KA
    Polymers (Basel), 2023 Nov 30;15(23).
    PMID: 38232019 DOI: 10.3390/polym15234594
    Oil-contaminated water and industrial oily wastewater discharges have adversely affected aquatic ecosystems and human safety. Membrane separation technology offers a promising solution for effective oil-water separation. Thus, a membrane with high surface area, hydrophilic-oleophobic properties, and stability is a promising candidate. Electrospinning, a straightforward and efficient process, produces highly porous polymer-based membranes with a vast surface area and stability. The main objective of this study is to produce hydrophilic-oleophobic polyacrylonitrile (PAN) and cellulose acetate (CA) nanofibers using core-shell electrospinning. Incorporating CA into the shell of the nanofibers enhances the wettability. The core PAN polymer improves the electrospinning process and contributes to the hydrophilicity-oleophobicity of the produced nanofibers. The PAN/CA nanofibers were characterized by Fourier transform infrared spectroscopy, field emission scanning electron microscopy, X-ray diffraction, and surface-wetting behavior. The resulting PAN/cellulose nanofibers exhibited significantly improved surface-wetting properties, demonstrating super-hydrophilicity and underwater superoleophobicity, making them a promising choice for oil-water separation. Various oils, including gasoline, diesel, toluene, xylene, and benzene, were employed in the preparation of oil-water mixture solutions. The utilization of PAN/CA nanofibers as a substrate proved to be highly efficient, confirming exceptional separation efficiency, remarkable stability, and prolonged durability. The current work introduces an innovative single-step fabrication method of composite nanofibers, specially designed for efficient oil-water separation. This technology exhibits significant promise for deployment in challenging situations, offering excellent reusability and a remarkable separation efficiency of nearly 99.9%.
  5. Fulltext Encapsulation of Bifidobacterium pseudocatenulatum Strain G4 within Bovine Gelatin-Genipin-Sodium Alginate Combinations: Optimisation Approach Using Face Central Composition Design-Response Surface Methodology (FCCD-RSM)
    Khalil KA, Mustafa S, Mohammad R, Ariff AB, Ahmad SA, Dahalan FA, et al.
    Int J Microbiol, 2019;2019:4208986.
    PMID: 31093290 DOI: 10.1155/2019/4208986
    Bovine gelatin is a biopolymer which has good potential to be used in encapsulating matrices for probiotic candidate Bifidobacterium pseudocatenulatum strain G4 (G4) because of its amphoteric nature characteristic. Beads were prepared by the extrusion method using genipin and sodium alginate as a cross-linking agent. The optimisation of bovine gelatin-genipin-sodium alginate combinations was carried out using face central composition design (FCCD) to investigate G4 beads' strength, before and after exposed to simulated gastric (SGF), intestinal fluids (SIF), and encapsulation yield. A result of ANOVA and the polynomial regression model revealed the combinations of all three factors have a significant effect (p < 0.05) on the bead strength. Meanwhile, for G4 encapsulation yield, only genipin showed less significant effect on the response. However, the use of this matrix remained due to the intermolecular cross-linking ability with bovine gelatin. Optimum compositions of bovine gelatin-genipin-sodium alginate were obtained at 11.21% (w/v), 1.96 mM, and 2.60% (w/v), respectively. A model was validated for accurate prediction of the response and showed no significant difference (p > 0.05) with experimental values.
  6. Autolysis of bovine skin, its endogenous proteases, protease inhibitors and their effects on quality characteristics of extracted gelatin
    Ahmad T, Ismail A, Ahmad SA, Khalil KA, Kee LT, Awad EA, et al.
    Food Chem, 2018 Nov 01;265:1-8.
    PMID: 29884359 DOI: 10.1016/j.foodchem.2018.05.046
    The autolysis of pretreated bovine skin (PBS) (treated with 0.1 M NaOH and 1% HCl), its endogenous proteases, inhibitors and their effects on quality attributes of gelatin were examined. PBS was subjected to different temperatures (20-90 °C) and pH (2-9) and treated with different protease inhibitors. Maximum autolytic activity of PBS was observed at 40 °C and pH 5. Ethylene-bis (oxyethylenenitrilo) tetraacetic acid (EGTA) was the most effective in impeding the degradation of γ-, β- and α- chains of PBS protein indicating that metallocollagenases were the predominant endogenous proteases in bovine skin. Gelatin was extracted in the absence (GAE) and presence (GPE) of EGTA, and EGTA with papain enzyme (GPEP). GPEP had a higher yield and lower gel strength than GEA and GPE. Metallocollagenases partook in the degradation of gelatin thereby affecting its functional properties. Pretreating PBS with or without EGTA, and papain influenced the quality attributes of gelatin.
  7. Fulltext Effects of Ultrasound Assisted Extraction in Conjugation with Aid of Actinidin on the Molecular and Physicochemical Properties of Bovine Hide Gelatin
    Ahmad T, Ismail A, Ahmad SA, Khalil KA, Leo TK, Awad EA, et al.
    Molecules, 2018 Mar 22;23(4).
    PMID: 29565325 DOI: 10.3390/molecules23040730
    Actinidin was used to pretreat the bovine hide and ultrasonic wave (53 kHz and 500 W) was used for the time durations of 2, 4 and 6 h at 60 °C to extract gelatin samples (UA2, UA4 and UA6, respectively). Control (UAC) gelatin was extracted using ultrasound for 6 h at 60 °C without enzyme pretreatment. There was significant (p < 0.05) increase in gelatin yield as the time duration of ultrasound treatment increased with UA6 giving the highest yield of 19.65%. Gel strength and viscosity of UAC and UA6 extracted gelatin samples were 627.53 and 502.16 g and 16.33 and 15.60 mPa.s, respectively. Longer duration of ultrasound treatment increased amino acids content of the extracted gelatin and UAC exhibited the highest content of amino acids. Progressive degradation of polypeptide chains was observed in the protein pattern of the extracted gelatin as the time duration of ultrasound extraction increased. Fourier transform infrared (FTIR) spectroscopy depicted loss of molecular order and degradation in UA6. Scanning electron microscopy (SEM) revealed protein aggregation and network formation in the gelatin samples with increasing time of ultrasound treatment. The study indicated that ultrasound assisted gelatin extraction using actinidin exhibited high yield with good quality gelatin.
  8. Fulltext Mathematical Modelling of Canola Oil Biodegradation and Optimisation of Biosurfactant Production by an Antarctic Bacterial Consortium Using Response Surface Methodology
    Zahri KNM, Khalil KA, Gomez-Fuentes C, Zulkharnain A, Sabri S, Convey P, et al.
    Foods, 2021 Nov 14;10(11).
    PMID: 34829082 DOI: 10.3390/foods10112801
    An Antarctic soil bacterial consortium (reference BS14) was confirmed to biodegrade canola oil, and kinetic studies on this biodegradation were carried out. The purpose of this study was to examine the ability of BS14 to produce biosurfactants during the biodegradation of canola oil. Secondary mathematical equations were chosen for kinetic analyses (Monod, Haldane, Teissier-Edwards, Aiba and Yano models). At the same time, biosurfactant production was confirmed through a preliminary screening test and further optimised using response surface methodology (RSM). Mathematical modelling demonstrated that the best-fitting model was the Haldane model for both waste (WCO) and pure canola oil (PCO) degradation. Kinetic parameters including the maximum degradation rate (μmax) and maximum concentration of substrate tolerated (Sm) were obtained. For WCO degradation these were 0.365 min-1 and 0.308%, respectively, while for PCO they were 0.307 min-1 and 0.591%, respectively. The results of all preliminary screenings for biosurfactants were positive. BS14 was able to produce biosurfactant concentrations of up to 13.44 and 14.06 mg/mL in the presence of WCO and PCO, respectively, after optimisation. The optimum values for each factor were determined using a three-dimensional contour plot generated in a central composite design, where a combination of 0.06% salinity, pH 7.30 and 1.55% initial substrate concentration led to the highest biosurfactant production when using WCO. Using PCO, the highest biosurfactant yield was obtained at 0.13% salinity, pH 7.30 and 1.25% initial substrate concentration. This study could help inform the development of large-scale bioremediation applications, not only for the degradation of canola oil but also of other hydrocarbons in the Antarctic by utilising the biosurfactants produced by BS14.
  9. Fulltext Antarctic heavy metal pollution and remediation efforts: state of the art of research and scientific publications
    Darham S, Zakaria NN, Zulkharnain A, Sabri S, Khalil KA, Merican F, et al.
    Braz J Microbiol, 2023 Sep;54(3):2011-2026.
    PMID: 36973583 DOI: 10.1007/s42770-023-00949-9
    In Antarctica, human activities have been reported to be the major cause of the accumulation of heavy metal contaminants. A comprehensive bibliometric analysis of publications on heavy metal contamination in Antarctica from year 2000 to 2020 was performed to obtain an overview of the current landscape in this line of research. A total of 106 documents were obtained from Scopus, the largest citation database. Extracted data were analysed, and VOSviewer software was used to visualise trends. The result showed an increase in publications and citations in the past 20 years indicating the rising interest on heavy metal contamination in the Antarctic region. Based on the analysis of keywords, the publications largely discuss various types of heavy metals found in the Antarctic water and sediment. The analysis on subject areas detects multiple disciplines involved, wherein the environmental science was well-represented. The top countries and authors producing the most publication in this field were from Australia, China, Brazil and Chile. Numerous efforts have been exercised to investigate heavy metal pollution and its mitigation approaches in the region in the past decades. This paper not only is relevant for scholars to understand the development status and trends in this field but also offers clear insights on the future direction of Antarctic heavy metal contamination and remediation research.
  10. Fulltext Managing Oxidative Stress Using Vitamin C to Improve Biocompatibility of Polycaprolactone for Bone Regeneration In Vitro
    Abdulhameed EA, Rani KGA, AlGhalban FM, Abou Neel EA, Khalifa N, Khalil KA, et al.
    ACS Omega, 2024 Jul 23;9(29):31776-31788.
    PMID: 39072128 DOI: 10.1021/acsomega.4c02858
    Increased oxidative stress in bone cells is known to negatively alter favorable bone regeneration. This study aimed to develop a porous polycaprolactone (PCL) membrane incorporated with 25 wt % Vitamin C (PCL-Vit C) and compared it to the PCL membrane to control oxidative stress and enhance biomineralization in vitro. Both membranes were characterized using SEM-EDS, FTIR spectroscopy, and surface hydrophilicity. Vitamin C release was quantified colorimetrically. Assessments of the viability and attachment of human fetal osteoblast (hFOB 1.19) cells were carried out using XTT assay, SEM, and confocal microscopy, respectively. ROS generation and wound healing percentage were measured using flow cytometry and ImageJ software, respectively. Mineralization study using Alizarin Red in the presence or absence of osteogenic media was carried out to measure the calcium content. Alkaline phosphatase assay and gene expression of osteogenic markers (alkaline phosphatase (ALP), collagen Type I (Col1), runt-related transcription factor 2 (RUNX2), osteocalcin (OCN), and osteopontin (OPN)) were analyzed by real-time PCR. SEM images revealed smooth, fine, bead-free fibers in both membranes. The FTIR spectrum of pure vitamin C was replaced with peaks at 3436.05 and 2322.83 cm-1 in the PCL-Vit C membrane. Vitamin C release was detected at 15 min and 1 h. The PCL-Vit C membrane was hydrophilic, generated lower ROS, and showed significantly higher viability than the PCL membrane. Although both PCL and PCL-Vit C membranes showed similar cellular and cytoskeletal morphology, more cell clusters were evident in the PCL-Vit C membrane. Lower ROS level in the PCL-Vit C membrane displayed improved cell functionality as evidenced by enhanced cellular differentiation with more intense alizarin staining and higher calcium content, supported by upregulation of osteogenic markers ALP, Col1, and OPN even in the absence of osteogenic supplements. The presence of Vitamin C in the PCL-Vit C membrane may have mitigated oxidative stress in hFOB 1.19 cells, resulting in enhanced biomineralization facilitating bone regeneration.
  11. A proposed aerobic granules size development scheme for aerobic granulation process
    Dahalan FA, Abdullah N, Yuzir A, Olsson G, Salmiati, Hamdzah M, et al.
    Bioresour Technol, 2015 Apr;181:291-6.
    PMID: 25661308 DOI: 10.1016/j.biortech.2015.01.062
    Aerobic granulation is increasingly used in wastewater treatment due to its unique physical properties and microbial functionalities. Granule size defines the physical properties of granules based on biomass accumulation. This study aims to determine the profile of size development under two physicochemical conditions. Two identical bioreactors namely Rnp and Rp were operated under non-phototrophic and phototrophic conditions, respectively. An illustrative scheme was developed to comprehend the mechanism of size development that delineates the granular size throughout the granulation. Observations on granules' size variation have shown that activated sludge revolutionised into the form of aerobic granules through the increase of biomass concentration in bioreactors which also determined the changes of granule size. Both reactors demonstrated that size transformed in a similar trend when tested with and without illumination. Thus, different types of aerobic granules may increase in size in the same way as recommended in the aerobic granule size development scheme.
  12. Fulltext Coco Peat as Agricultural Waste Sorbent for Sustainable Diesel-Filter System
    Verasoundarapandian G, Zakaria NN, Shaharuddin NA, Khalil KA, Puasa NA, Azmi AA, et al.
    Plants (Basel), 2021 Nov 16;10(11).
    PMID: 34834831 DOI: 10.3390/plants10112468
    Oil spill incidents are hazardous and have prolonged damage to the marine environment. Management and spill clean-up procedures are practical and rapid, with several shortcomings. Coco peat (CP) and coco fibre (CF) are refined from coconut waste, and their abundance makes them desirable for diesel spillage treatment. Using a filter-based system, the selectivity of coco peat sorbent was tested using CP, CF and peat-fibre mix (CPM). CP exhibited maximal diesel sorption capacity with minimal seawater uptake, thus being selected for further optimisation analysis. The heat treatment considerably improved the sorption capacity and efficiency of diesel absorbed by CP, as supported by FTIR and VPSEM-EDX analysis. Conventional one-factor-at-a-time (OFAT) examined the performance of diesel sorption by CP under varying parameters, namely temperature, time of heating, packing density and diesel concentration. The significant factors were statistically evaluated using response surface methodology (RSM) via Plackett-Burman design (PB) and central composite design (CCD). Three significant (p < 0.05) factors (time, packing density and diesel concentration) were identified by PB and further analysed for interactions among the parameters. CCD predicted efficiency of diesel absorbed at 59.92% (71.90 mL) (initial diesel concentration of 30% v/v) and the experimental model validated the design with 59.17% (71.00 mL) diesel sorbed at the optimised conditions of 14.1 min of heating (200 °C) with packing density of 0.08 g/cm3 and 30% (v/v) of diesel concentration. The performance of CP in RSM (59.17%) was better than that in OFAT (58.33%). The discoveries imply that natural sorbent materials such as CP in oil spill clean-up operations can be advantageous and environmentally feasible. This study also demonstrated the diesel-filter system as a pilot study for the prospective up-scale application of oil spills.
  13. Effects of heavy metals on bacterial growth parameters in degradation of phenol by an Antarctic bacterial consortium
    Tengku-Mazuki TA, Darham S, Convey P, Shaharuddin NA, Zulkharnain A, Khalil KA, et al.
    Braz J Microbiol, 2024 Mar;55(1):629-637.
    PMID: 38110706 DOI: 10.1007/s42770-023-01215-8
    Antarctica has often been perceived as a pristine continent until the recent few decades as pollutants have been observed accruing in the Antarctic environment. Irresponsible human activities such as accidental oil spills, waste incineration and sewage disposal are among the primary anthropogenic sources of heavy metal contaminants in Antarctica. Natural sources including animal excrement, volcanism and geological weathering also contribute to the increase of heavy metals in the ecosystem. A microbial growth model is presented for the growth of a bacterial cell consortium used in the biodegradation of phenol in media containing different metal ions, namely arsenic (As), cadmium (Cd), aluminium (Al), nickel (Ni), silver (Ag), lead (Pb) and cobalt (Co). Bacterial growth was inhibited by these ions in the rank order of Al 
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