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  1. A novel chitinous nanoparticles prepared and characterized with black soldier fly (Hermetia illucens L.) using steam flash explosion treatment
    Feng H, Wang Z, Sajab MS, Abdul PM, Ding G
    Int J Biol Macromol, 2023 Jan 10;230:123210.
    PMID: 36639077 DOI: 10.1016/j.ijbiomac.2023.123210
    This is the first report of the use of steam flash explosion (SFE) to prepare chitinous nanoparticles from black soldier fly (BSF). SFE treatment was performed at a steam pressure of 0.45 to 1.60 MPa with a holding time of 60 s. As the pressure increased, the particle size of the chitinous particles decreased. Under SFE at 1.60 MPa, chitinous nanoparticles with sizes ranging from 59 to 162 nm were produced. SEM, AFM, Raman spectroscopy, FT-IR spectroscopy, 1H NMR, TGA, and DSC were used to characterize the BSF chitin materials. It was demonstrated that SFE treatment deacetylated chitin to obtain chitosan with 91.24 % deacetylation. In addition, the polymer backbone was maintained, and the degree of polymerization of chitosan nanoparticles was reduced. The activity of the cationic groups of chitosan nanoparticles was improved, thereby enhancing the temperature sensitivity of the polymeric material. It can be concluded that the SFE one-step processing method is a simple and efficient way to prepare homogeneous biomaterial nanoparticles. This study has implications for the development of chitosan nanomaterials for biomedical applications.
  2. Two-Stage and One-Stage Anaerobic Co-digestion of Vinasse and Spent Brewer Yeast Cells for Biohydrogen and Methane Production
    Nualsri C, Abdul PM, Imai T, Reungsang A, Sittijunda S
    Mol Biotechnol, 2024 Jan 17.
    PMID: 38231316 DOI: 10.1007/s12033-023-01015-3
    This study aimed to evaluate the two-stage and one-stage anaerobic co-digestion of vinasse and spent brewer yeast cells (SBY) for biohydrogen and methane production. Optimization of the vinasse-to-SBY ratio and fly ash concentration of the two-stage and one-stage production processes was investigated. In the two-stage process, the vinasse-to-SBY ratio and fly ash concentration were optimized, and the leftover effluent was used for methane production. The optimum conditions for biohydrogen production were a vinasse-to-SBY ratio of 7:3% v/w and fly ash concentration of 0.4% w/v, in which the maximum hydrogen yield was 43.7 ml-H2/g-VSadded. In contrast, a vinasse-to-SBY ratio of 10:0% v/w and fly ash concentration of 0.2% w/v were considered optimal for methane production, and resulted in a maximum methane yield of 214.6 ml-CH4/g-VSadded. For the one-stage process, a vinasse-to-SBY ratio of 10:0% v/w and fly ash concentration of 0.1% w/v were considered optimal, and resulted in a maximum methane yield of 243.6 ml-CH4/g-VSadded. In the two-stage process, the energy yield from hydrogen (0.05-0.47 kJ/g-VSadded) was 0.62%-11.78%, and the major fraction was approximately 88.22%-99.38% gain from methane (3.19-7.73 kJ/g-VSadded). For the one-stage process, the total energy yield distribution ranged from 4.20 to 8.77 kJ/g-VSadded.
  3. Compatibility of utilising nitrogen-rich oil palm trunk sap for succinic acid fermentation by Actinobacillus succinogenes 130Z
    Bukhari NA, Loh SK, Nasrin AB, Luthfi AAI, Harun S, Abdul PM, et al.
    Bioresour Technol, 2019 Dec;293:122085.
    PMID: 31499328 DOI: 10.1016/j.biortech.2019.122085
    In this study, the potential of oil palm trunk (OPT) sap as a sole substrate for succinic acid (SA) production was evaluated using Actinobacillus succinogenes 130Z. After OPT sap was characterised, the effects of adding carbonate, yeast extract (YE) and minerals to this medium were investigated in an attempt to develop a low-cost fermentation medium. The OPT sap alone, gave comparable SA yield and productivity (0.54 g/g and 0.35 g/L/h) to those supplemented with YE (0.50 g/g and 0.36 g/L/h) and minerals (0.55 g/g and 0.40 g/L/h). The findings showed that OPT sap has sufficient amount of nutrients for SA biosynthesis by A. succinogenes 130Z and could potentially reduce cost without requiring expensive nutrients supplementation.
  4. Low cost nutrient-rich oil palm trunk bagasse hydrolysate for bio-succinic acid production by Actinobacillus succinogenes
    Bukhari NA, Loh SK, Luthfi AAI, Abdul PM, Jahim JM
    Prep Biochem Biotechnol, 2021 Dec 22.
    PMID: 34935581 DOI: 10.1080/10826068.2021.2015692
    Economical source of succinic acid (SA) is most sought-after as a key platform chemical for a wide range of applications. Low-cost production of bio-succinic acid (bio-SA) from a renewable biomass resource i.e., oil palm trunk (OPT) is reported in this paper. Apart from carbon source, nitrogen source and mineral salts are other important nutrients affecting microbial cell growth and bio-SA biosynthesis by Actinobacillus succinogenes 130Z. In order to access and optimize nutrient requirement of the latter two sources, their effects in terms of types and concentrations were investigated. The findings highlighted the importance of selecting proper nitrogen source in A. succinogenes fermentation. The possibility of producing bio-SA from OPT economically can be achieved through minimal supply of 5 g/L yeast extract compared to that generally supplemented 15 g/L with a similar yield (0.47 g/g). In addition, a higher bio-SA yield (0.49 g/g) was achieved without adding mineral salts, which could further reduce fermentation cost. The use of minimally supplemented hydrolysate resulted in 21.1 g/L of bio-SA with a satisfactory yield (0.58 g/g) in a batch bioreactor system with an estimated 56.4% in cost savings. Conclusively, OPT bagasse hydrolysate is a nutrient-rich feedstock that can be practically utilized for bio-SA production.
  5. Fulltext Classification of oil palm fresh fruit maturity based on carotene content from Raman spectra
    Raj T, Hashim FH, Huddin AB, Hussain A, Ibrahim MF, Abdul PM
    Sci Rep, 2021 09 15;11(1):18315.
    PMID: 34526627 DOI: 10.1038/s41598-021-97857-5
    The oil yield, measured in oil extraction rate per hectare in the palm oil industry, is directly affected by the ripening levels of the oil palm fresh fruit bunches at the point of harvesting. A rapid, non-invasive and reliable method in assessing the maturity level of oil palm harvests will enable harvesting at an optimum time to increase oil yield. This study shows the potential of using Raman spectroscopy to assess the ripeness level of oil palm fruitlets. By characterizing the carotene components as useful ripeness features, an automated ripeness classification model has been created using machine learning. A total of 46 oil palm fruit spectra consisting of 3 ripeness categories; under ripe, ripe, and over ripe, were analyzed in this work. The extracted features were tested with 19 classification techniques to classify the oil palm fruits into the three ripeness categories. The Raman peak averaging at 1515 cm-1 is shown to be a significant molecular fingerprint for carotene levels, which can serve as a ripeness indicator in oil palm fruits. Further signal analysis on the Raman peak reveals 4 significant sub bands found to be lycopene (ν1a), β-carotene (ν1b), lutein (ν1c) and neoxanthin (ν1d) which originate from the C=C stretching vibration of carotenoid molecules found in the peel of the oil palm fruit. The fine KNN classifier is found to provide the highest overall accuracy of 100%. The classifier employs 6 features: peak intensities of bands ν1a to ν1d and peak positions of bands ν1c and ν1d as predictors. In conclusion, the Raman spectroscopy method has the potential to provide an accurate and effective way in determining the ripeness of oil palm fresh fruits.
  6. Biotechnological enhancement of lactic acid conversion from pretreated palm kernel cake hydrolysate by Actinobacillus succinogenes 130Z
    Rahim NA, Luthfi AAI, Bukhari NA, Tan JP, Abdul PM, Manaf SFA
    Sci Rep, 2023 Apr 08;13(1):5787.
    PMID: 37031272 DOI: 10.1038/s41598-023-32964-z
    The aim of this study was to establish an improved pretreatment and fermentation method i.e. immobilized cells for high recovery of fermentable sugars from palm kernel cake (PKC) and its effects on fermentability performance by Actinobacillus succinogenes 130Z in the conversion of the fermentable sugar to lactic acid. The effects of oxalic acid concentrations (1-6% w/v) and residence times (1-5 h) on the sugar recovery were initially investigated and it was found that the highest mannose concentration was 25.1 g/L at the optimum hydrolysis conditions of 4 h and 3% (w/v) oxalic acid. The subsequent enzymatic saccharification of the pretreated PKC afforded the highest enzymatic digestibility with the recovered sugars amounting to 25.18 g/L and 9.14 g/L of mannose and glucose, respectively. Subsequently, the fermentability performance of PKC hydrolysate was evaluated and compared in terms of cultivation phases (i.e. mono and dual-phases), carbonate loadings (i.e. magnesium and sodium carbonates), and types of sugars (i.e. glucose and mannose). The highest titer of 19.4 g/L lactic acid was obtained from the fermentation involving A. succinogenes 130Z in dual-phase cultivation supplemented with 30 g/L of magnesium carbonate. Lactic acid production was further enhanced by using immobilized cells with coconut shell-activated carbon (CSAC) of different sizes (A, B, C, and D) in the repeated batch cultivation of dual-phase fermentation producing 31.64 g/L of lactic acid. This work sheds light on the possibilities to enhance the utilization of PKC for lactic acid production via immobilized A. succinogenes 130Z.
  7. Towards consolidated bioprocessing of biomass and plastic substrates for semi-synthetic production of bio-poly(ethylene furanoate) (PEF) polymer using omics-guided construction of artificial microbial consortia
    Omar MN, Minggu MM, Nor Muhammad NA, Abdul PM, Zhang Y, Ramzi AB
    Enzyme Microb Technol, 2024 Jun;177:110429.
    PMID: 38537325 DOI: 10.1016/j.enzmictec.2024.110429
    Poly(ethylene furanoate) (PEF) plastic is a 100% renewable polyester that is currently being pursued for commercialization as the next-generation bio-based plastic. This is in line with growing demand for circular bioeconomy and new plastics economy that is aimed at minimizing plastic waste mismanagement and lowering carbon footprint of plastics. However, the current catalytic route for the synthesis of PEF is impeded with technical challenges including high cost of pretreatment and catalyst refurbishment. On the other hand, the semi-biosynthetic route of PEF plastic production is of increased biotechnological interest. In particular, the PEF monomers (Furan dicarboxylic acid and ethylene glycol) can be synthesized via microbial-based biorefinery and purified for subsequent catalyst-mediated polycondensation into PEF. Several bioengineering and bioprocessing issues such as efficient substrate utilization and pathway optimization need to be addressed prior to establishing industrial-scale production of the monomers. This review highlights current advances in semi-biosynthetic production of PEF monomers using consolidated waste biorefinery strategies, with an emphasis on the employment of omics-driven systems biology approaches in enzyme discovery and pathway construction. The roles of microbial protein transporters will be discussed, especially in terms of improving substrate uptake and utilization from lignocellulosic biomass, as well as from depolymerized plastic waste as potential bio-feedstock. The employment of artificial bioengineered microbial consortia will also be highlighted to provide streamlined systems and synthetic biology strategies for bio-based PEF monomer production using both plant biomass and plastic-derived substrates, which are important for circular and new plastics economy advances.
  8. Effects of changes in chemical and structural characteristic of ammonia fibre expansion (AFEX) pretreated oil palm empty fruit bunch fibre on enzymatic saccharification and fermentability for biohydrogen
    Abdul PM, Jahim JM, Harun S, Markom M, Lutpi NA, Hassan O, et al.
    Bioresour Technol, 2016 Jul;211:200-8.
    PMID: 27017130 DOI: 10.1016/j.biortech.2016.02.135
    Oil palm empty fruit bunch (OPEFB) fibre is widely available in Southeast Asian countries and found to have 60% (w/w) sugar components. OPEFB was pretreated using the ammonia fibre expansion (AFEX) method and characterised physically by the Fourier transform infrared spectroscopy, X-ray diffraction and scanning electron microscopy. The results show that there were significant structural changes in OPEFB after the pretreatment step, and the sugar yield after enzymatic hydrolysis using a cocktail of Cellic Ctec2® and Cellic Htec2® increased from 0.15gg(-1) of OPEFB in the raw untreated OPEFB sample to 0.53gg(-1) of OPEFB in AFEX-pretreated OPEFB (i.e. almost a fourfold increase in sugar conversion), which enhances the economic value of OPEFB. A biohydrogen fermentability test of this hydrolysate was carried out using a locally isolated bacterium, Enterobacter sp. KBH6958. The biohydrogen yield after 72h of fermentation was 1.68mol H2 per mol sugar. Butyrate, ethanol, and acetate were the major metabolites.
  9. Dose-response analysis of toxic effect from palm oil mill effluent (POME) by-products on biohydrogen producing bacteria - A preliminary study on microbial density and determination of EC50
    Arisht SN, Abdul PM, Jasni J, Mohd Yasin NH, Lin SK, Wu SY, et al.
    Ecotoxicol Environ Saf, 2020 Oct 15;203:110991.
    PMID: 32888602 DOI: 10.1016/j.ecoenv.2020.110991
    The stimulant and toxicity effects of reported organic (acetic acid, propionic acid, butyric acid, formic acid, oil & grease) and inorganic (copper) by-products presented in palm oil mill effluent on anaerobic bacterial population were examined in this paper. The toxicity test had shown that acetic, propionic and butyric acids tend to stimulate the bacterial density level (survival rate more than 50%), while formic acid, copper, oil and grease were shown to have suppressed the density level (survival rate less than 50%). The highest biomass recorded was 1.66 mg/L for the concentration of acetic acid at 216 mg/L and lowest biomass concentration, 0.90 mg/L for copper at 1.40 mg/L. Biohydrogen-producing bacteria have a favourable growth rate around pH 5.5. The comparison of half maximal effective concentration (EC50) values between two test duration on the effects of organic and inorganic by-products postulate that bacteria had a higher tolerance towards volatile fatty acids. While acetic, butyric and propionic acids had exhibited higher tolerance EC50 values for bacteria, but the opposite trend was observed for formic acid, copper and oil & grease.
  10. Fulltext Zinc removal and recovery from industrial wastewater with a microbial fuel cell: Experimental investigation and theoretical prediction
    Lim SS, Fontmorin JM, Pham HT, Milner E, Abdul PM, Scott K, et al.
    Sci Total Environ, 2021 Jul 01;776:145934.
    PMID: 33647656 DOI: 10.1016/j.scitotenv.2021.145934
    Microbial fuel cells (MFCs) that simultaneously remove organic contaminants and recovering metals provide a potential route for industry to adopt clean technologies. In this work, two goals were set: to study the feasibility of zinc removal from industrial effluents using MFCs and to understand the removal process by using reaction rate models. The removal of Zn2+ in MFC was over 96% for synthetic and industrial samples with initial Zn2+ concentrations less than 2.0 mM after 22 h of operation. However, only 83 and 42% of the zinc recovered from synthetic and industrial samples, respectively, was attached on the cathode surface of the MFCs. The results marked the domination of electroprecipitation rather than the electrodeposition process in the industrial samples. Energy dispersive X-ray (EDX) analysis showed that the recovered compound contained not only Zn but also O, evidence that Zn(OH)2 could be formed. The removal of Zn2+ in the MFC followed a mechanism where oxygen was reduced to hydroxide before reacting with Zn2+. Nernst equations and rate law expressions were derived to understand the mechanism and used to estimate the Zn2+ concentration and removal efficiency. The zero-, first- and second-order rate equations successfully fitted the data, predicted the final Zn2+ removal efficiency, and suggested that possible mechanistic reactions occurred in the electrolysis cell (direct reduction), MFC (O2 reduction), and control (chemisorption) modes. The half-life, t1/2, of the Zn2+ removal reaction using synthetic and industrial samples was estimated to be 7.0 and 2.7 h, respectively. The t1/2 values of the controls (without the power input from the MFC bioanode) were much slower and were recorded as 21.5 and 7.3 h for synthetic and industrial samples, respectively. The study suggests that MFCs can act as a sustainable and environmentally friendly technology for heavy metal removal without electrical energy input or the addition of chemicals.
  11. Fulltext An alternative peptone preparation using Hermetia illucens (Black soldier fly) hydrolysis: process optimization and performance evaluation
    Liu G, Tiang MF, Ma S, Wei Z, Liang X, Sajab MS, et al.
    PeerJ, 2024;12:e16995.
    PMID: 38426145 DOI: 10.7717/peerj.16995
    BACKGROUND: Hermetia illucens (HI), commonly known as the black soldier fly, has been recognized for its prowess in resource utilization and environmental protection because of its ability to transform organic waste into animal feed for livestock, poultry, and aquaculture. However, the potential of the black soldier fly's high protein content for more than cheap feedstock is still largely unexplored.

    METHODS: This study innovatively explores the potential of H. illucens larvae (HIL) protein as a peptone substitute for microbial culture media. Four commercial proteases (alkaline protease, trypsin, trypsase, and papain) were explored to hydrolyze the defatted HIL, and the experimental conditions were optimized via response surface methodology experimental design. The hydrolysate of the defatted HIL was subsequently vacuum freeze-dried and deployed as a growth medium for three bacterial strains (Staphylococcus aureus, Bacillus subtilis, and Escherichia coli) to determine the growth kinetics between the HIL peptone and commercial peptone.

    RESULTS: The optimal conditions were 1.70% w/w complex enzyme (alkaline protease: trypsin at 1:1 ratio) at pH 7.0 and 54 °C for a duration of 4 h. Under these conditions, the hydrolysis of defatted HIL yielded 19.25% ±0.49%. A growth kinetic analysis showed no significant difference in growth parameters (μmax, Xmax, and λ) between the HIL peptone and commercial peptone, demonstrating that the HIL hydrolysate could serve as an effective, low-cost alternative to commercial peptone. This study introduces an innovative approach to HIL protein resource utilization, broadening its application beyond its current use in animal feed.

  12. Fulltext Potential Utilisation of Dark-Fermented Palm Oil Mill Effluent in Continuous Production of Biomethane by Self-Granulated Mixed Culture
    Mahmod SS, Azahar AM, Luthfi AAI, Abdul PM, Mastar MS, Anuar N, et al.
    Sci Rep, 2020 Jun 08;10(1):9167.
    PMID: 32514030 DOI: 10.1038/s41598-020-65702-w
    Two-stage anaerobic digestion of palm oil mill effluent (POME) is a promising method for converting the waste from the largest agricultural industry in Southeast Asia into a clean and sustainable energy. This study investigates the degradation of acid-rich effluent from the dark fermentation stage for the production of biomethane (BioCH4) in a 30-L continuous stirred-tank reactor (CSTR). The continuous methanogenic process was operated with varied HRTs (10 - 1 day) and OLRs (4.6-40.6 gCOD/L.d-1) under thermophilic conditions. Methanothermobacter sp. was the dominant thermophilic archaea that was responsible for the production rate of 4.3 LCH4/LPOME.d-1 and methane yield of 256.77 LCH4kgCOD at HRT of 2 d, which is the lowest HRT reported in the literature. The process was able to digest 85% and 64% of the initial POME's COD and TSS, respectively. The formation of methane producing granules (MPG) played a pivotal role in sustaining the efficient and productive anaerobic system. We report herein that the anaerobic digestion was not only beneficial in reducing the contaminants in the liquid effluent, but generating BioCH4 gas with a positive net energy gain of 7.6 kJ/gCOD.
  13. Precisely controlled electrostatically sprayed sodium alginate/carboxymethyl chitosan hydrogel microbeads as super-adsorbent for adsorption of cationic dye
    Li X, Tiang MF, Cui X, Li Y, Wang Z, Zhao L, et al.
    Int J Biol Macromol, 2024 Nov 22;283(Pt 4):137989.
    PMID: 39581417 DOI: 10.1016/j.ijbiomac.2024.137989
    In this pioneering study, electrostatic spraying (ES) technology with high voltages is proposed to reduce the size of hydrogel microbeads further, aiming to enhance the adsorption rate of cationic methylene blue (MB) dye. The increased voltages, ranging from 0.0 to 13.0 kV, further decreased the size of electrostatically sprayed hydrogel microbeads crosslinked by hydrogen bonds between sodium alginate (SA) and carboxymethyl chitosan (CMCS) in hydrochloric acid. The size of SA/CMCS hydrogel microbeads was successfully reduced from 2000 ± 121 μm (SC-2000) to 400 ± 15 μm (SC-400). Notably, SC-400 exhibits the highest maximum adsorption capacity (qm) and rate constant (k2) at 840.3 mg/g and 0.0598 g/mg/min, respectively, at pH 9.0 and a temperature of 25 °C in the absence of ionic compounds, which is three times higher than that of SC-2000, due to their high specific surface area and pore volume. Through a series of adsorption studies and characterization analyses, SA/CMCS hydrogel microbeads displayed heterogeneous adsorption behaviors towards MB dye through electrostatic interactions between the deprotonated carboxylic groups and cationic MB molecules, where MB adsorption efficiency could be significantly influenced by pH and ionic strength. These findings suggest that ES technology is effective in synthesizing smaller SA/CMCS hydrogel microbeads with enhanced MB removal rates and stable adsorption capacities and their applications could be further explored for removing other organic dyes and toxic metals in subsequent research studies.
  14. Impact of light spectra on photo-fermentative biohydrogen production by Rhodobacter sphaeroides KKU-PS1
    Tiang MF, Hanipa MAF, Mahmod SS, Zainuddin MT, Lutfi AAI, Jahim JM, et al.
    Bioresour Technol, 2024 Feb;394:130222.
    PMID: 38109981 DOI: 10.1016/j.biortech.2023.130222
    Purple non-sulphur bacteria can only capture up to 10 % light spectra and only 1-5 % of light is converted efficiently for biohydrogen production. To enhance light capture and conversion efficiencies, it is necessary to understand the impact of various light spectra on light harvesting pigments. During photo-fermentation, Rhodobacter sphaeroides KKU-PS1 cultivated at 30 °C and 150 rpm under different light spectra has been investigated. Results revealed that red light is more beneficial for biomass accumulation, whereas green light showed the greatest impact on photo-fermentative biohydrogen production. Light conversion efficiency by green light is 2-folds of that under control white light, hence photo-hydrogen productivity is ranked as green > red > orange > violet > blue > yellow. These experimental data demonstrated that green and red lights are essential for photo-hydrogen and biomass productions of R. sphaeroides and a clearer understanding that possibly pave the way for further photosynthetic enhancement research.
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