Enhancing biohydrogen gas production in anaerobic system via comparative chemical pre-treatment on palm oil mill effluent (POME).

Biological hydrogen production using palm oil mill effluent (POME) as a carbon source through dark fermentation process has been suggested to be a promising bioenergy potential and enacts as alternative renewable energy source. Results have indicated that among various 1.5% (w/v) chemical pre-treatments (sodium hydroxide, NaOH; hydrochloric acid, HCl; sulphuric acid, H2SO4; phosphoric acid, H3PO4 and nitric acid, HNO3) on POME, using H3PO4 would generate maximum biohydrogen production of 0.193 mmol/L/h, which corresponded to a yield of 1.51 mol H2/mol TCconsumed with an initial total soluble carbohydrate concentration of 23.52 g/L. Among H3PO4 concentrations (1%-10%), the soluble carbohydrate content and the biohydrogen produced was highest and increased by 1.70-fold and 2.35-fold respectively at 2.5% (w/v), as compared to untreated POME. The batch fermentation maximum hydrogen production rate and yield of 0.208 mmol/L/h and 1.69 mol H2/mol TCconsumed were achieved at optimum pre-treatment conditions of pH 5.5 and thermophilic temperature (60 °C). This study suggests that chemical pre-treatment approaches manage to produce and improve the carbohydrate utilisation process further. Continuous fermentation in CSTR at the optimum conditions produce heightened 1.5-fold biohydrogen yield for 2.5% H3PO4 at 6 h HRT as compared to batch scale. Bacterial community via next-generation sequencing analysis at optimum HRT (6 h) revealed that Thermoanaerobacterium thermosaccharolyticum registered the highest relative frequency of 20.24%. At the class level, Clostridia, Bacilli, Bacteroidia, Thermoanaerobacteria, and Gammaproteobacteria were identified as the biohydrogen-producing bacteria in the continuous system. Insightful findings from this study suggest the substantial practical utility of dilute chemical pre-treatment in improving biohydrogen production.

Low cost nutrient-rich oil palm trunk bagasse hydrolysate for bio-succinic acid production by Actinobacillus succinogenes.

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.

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.

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.

Two-Stage and One-Stage Anaerobic Co-digestion of Vinasse and Spent Brewer Yeast Cells for Biohydrogen and Methane Production.

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.

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.

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.

An alternative peptone preparation using Hermetia illucens (Black soldier fly) hydrolysis: process optimization and performance evaluation.

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.

Impact of light spectra on photo-fermentative biohydrogen production by Rhodobacter sphaeroides KKU-PS1.

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.

A novel chitinous nanoparticles prepared and characterized with black soldier fly (Hermetia illucens L.) using steam flash explosion treatment.

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.

Nanocellulose-Based Biomaterial Ink Hydrogel for Uptake/Release of Bovine Serum Albumin.

This study explores the potential of using nanocellulose extracted from oil palm empty fruit bunch (OPEFB) as a biomaterial ink for 3D printing. The research focuses on using nanocellulose hydrogels for the controlled uptake and release of proteins, with the specific protein solution being Bovine Serum Albumin (BSA). To provide a suitable material for the bioprinting process, the study examines the characteristics and properties of the printed hydrogels through various analyses, such as morphology, functional group, crystallinity, and compression test. Several parameters, such as initial concentration, temperature, and the presence of calcium chloride as an additional crosslinker, affect the protein uptake and release capabilities of the hydrogel. The study is important for biomedicine as it explores the behavior of protein uptake and release using nanocellulose and 3D printing and can serve as a preliminary study for using hydrogels in biological materials or living cells.

Biotechnological enhancement of lactic acid conversion from pretreated palm kernel cake hydrolysate by Actinobacillus succinogenes 130Z.

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.

Classification of oil palm fresh fruit maturity based on carotene content from Raman spectra.

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), ß-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.

Zinc removal and recovery from industrial wastewater with a microbial fuel cell: Experimental investigation and theoretical prediction.

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.

Potential Utilisation of Dark-Fermented Palm Oil Mill Effluent in Continuous Production of Biomethane by Self-Granulated Mixed Culture.

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.

Compatibility of utilising nitrogen-rich oil palm trunk sap for succinic acid fermentation by Actinobacillus succinogenes 130Z.

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.

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.

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.