Effect of oil palm biomass cellulosic content on nanopore structure and adsorption capacity of biochar.

The influence of biomass cellulosic content on biochar nanopore structure and adsorption capacity in aqueous phase was scarcely reported. Commercial cellulose (100% cellulose), oil palm frond (39.5% cellulose), and palm kernel shell (20.5% cellulose) were pyrolyzed AT 630 °C, characterized and tested for the adsorption of iodine and organic contaminants. The external surface area and average pore size increased with cellulosic content, where commercial cellulose formed biochar with external surface area of 95.4 m2/g and average pore size of 4.1 nm. The biochar from commercial cellulose had the largest adsorption capacities: 371.40 mg/g for iodine, 86.7 mg/L for tannic acid, 17.89 mg/g for COD and 60.35 mg/g for colour, while biochar from palm kernel shell had the least adsorption capacities. The cellulosic content reflected the differences in biochar nanopore structure and adsorption capacities, signifying the suitability of highly cellulosic biomass for producing biochar to effectively treat wastewater.

Valorization of biodiesel side stream waste glycerol for rhamnolipids production by Pseudomonas aeruginosa RS6.

Biodiesel side stream waste glycerol was identified as a cheap carbon source for rhamnolipids (RLs) production which at the same time could improve the management of waste. The present study aimed to produce RLs by using Pseudomonas aeruginosa RS6 utilizing waste glycerol as a substrate and to evaluate their physico-chemicals properties. Fermentation conditions such as temperature, initial medium pH, waste glycerol concentration, nitrogen sources and concentrations resulted in different compositions of the mono- and di-RLs produced. The maximum RLs production of 2.73 g/L was obtained when P. aeruginosa RS6 was grown in a basal salt medium supplemented with 1% waste glycerol and 0.2 M sodium nitrate at 35 °C and pH 6.5. At optimal fermentation conditions, the emulsification index (E24) values of cooking oil, diesel oil, benzene, olive oil, petroleum, and kerosene were all above E24=50%. The surface tension reduction obtained from 72.13 mN/m to 29.4-30.4 mN/m was better than the surface activity of some chemical-based surfactants. The RLs produced possessed antimicrobial activities against Gram-negative and Gram-positive bacteria with values ranging from 37% to 77% of growth inhibition when 1 mg/mL of RLs was used. Concentrations of RLs below 1500 µg/mL did not induce phytotoxicity effects on the tested seeds (Vigna radiata) compared to the chemical-based- surfactant, SDS. Furthermore, RLs tested on zebrafish (Danio rerio) embryos only exhibited low acute toxicity with an LC50 value of 72.97 µg/mL at 48 h of exposure suggesting a green and eco-biochemical worthy of future applications to replace chemical-based surfactants.

Ecotoxicological assessment of palm oil mill effluent final discharge by zebrafish (Danio rerio) embryonic assay.

Most palm oil mills adopted conventional ponding system, including anaerobic, aerobic, facultative and algae ponds, for the treatment of palm oil mill effluent (POME). Only a few mills installed a bio-polishing plant to treat POME further before its final discharge. The present study aims to determine the quality and toxicity levels of POME final discharge from three different mills by using conventional chemical analyses and fish (Danio rerio) embryo toxicity (FET) test. The effluent derived from mill A which installed with a bio-polishing plant had lower values of BOD, COD and TSS at 45 mg/L, 104 mg/L, and 27 mg/L, respectively. Only mill A nearly met the industrial effluent discharge standard for BOD. In FET test, effluent from mill A recorded low lethality and most of the embryos were malformed after hatching (half-maximal effective concentration (EC50) = 20%). The highest toxicity was observed from the effluent of mill B and all embryos were coagulated after 24 h in samples greater than 75% of effluent (38% of half-maximal lethal concentration (LC50) at 96 h). The embryos in the effluent from mill C recorded high mortality after hatching, and the survivors were malformed after 96 h exposure (LC50 = 26%). Elemental analysis of POME final discharge samples showed Cu, Zn, and Fe concentrations were in the range of 0.10-0.32 mg/L, 0.01-0.99 mg/L, and 0.94-4.54 mg/L, respectively and all values were below the effluent permissible discharge limits. However, the present study found these metals inhibited D. rerio embryonic development at 0.12 mg/L of Cu, and 4.9 mg/L of Fe for 96 h-EC50. The present study found that bio-polishing plant installed in mill A effectively removing pollutants especially BOD and the FET test was a useful method to monitor quality and toxicity of the POME final discharge samples.

Adsorption mechanism and effectiveness of phenol and tannic acid removal by biochar produced from oil palm frond using steam pyrolysis.

In order to meet the growing demand for adsorbents to treat wastewater effectively, there has been increased interest in using sustainable biomass feedstocks. In this present study, the dermal tissue of oil palm frond was pyrolyzed with superheated steam at 500 °C to produce nanoporous biochar as bioadsorbent. The effect of operating conditions was investigated to understand the adsorption mechanism and to enhance the adsorption of phenol and tannic acid. The biochar had a microporous structure with a Brunauer-Emmett-Teller surface area of 422 m2/g containing low polar groups. The adsorption capacity of 62.89 mg/g for phenol and 67.41 mg/g for tannic acid were obtained using 3 g/L biochar dosage after 8 h of treatment at solution pH of 6.5 and temperature of 45 °C. The Freundlich model had the best fit to the isotherm data of phenol (R2 of 0.9863), while the Langmuir model best elucidated the isotherm data of tannic acid (R2 of 0.9632). These indicated that the biochar-phenol interface was associated with a heterogeneous multilayer sorption mechanism, while the biochar-tannic acid interface had a nonspecific monolayer sorption mechanism. The residual concentration of 26.3 mg/L phenol and 23.1 mg/L tannic acid was achieved when treated from 260 mg/L three times consecutively with 1 g/L biochar dosage, compared to a reduction to 72.3 mg/L phenol and 69.9 mg/L tannic acid using 3 g/L biochar dosage in a single treatment. The biochar exhibited effective adsorption of phenol and tannic acid, making it possible to treat effluents that contain varieties of phenolic compounds.

Toxicity identification and evaluation of palm oil mill effluent and its effects on the planktonic crustacean Daphnia magna.

Palm oil mill effluent (POME) contains complex and highly biodegradable organic matters so discharging it without appropriate treatment may lead to environmental problems. POME final discharge quality is normally determined based on conventional chemical detection such as by biological oxygen demand (BOD) and chemical oxygen demand (COD). The novelty of the present study is that the toxicity effects of the POME final discharge samples were evaluated based on whole effluent toxicity (WET) and toxicity identification evaluation (TIE) tests using Daphnia magna. The toxicity unit (TU) values were recorded to be in the range from TU = 1.1-11 obtained from WET, and the TIE manipulation tests suggested that a substantial amount of toxic compounds was contained in the POME final discharge. Phenol, 2,6-bis (1,1-dimethylethyl)- and heavy metals such as Cu and Zn were detected in all the effluents and were recognized as being the main toxicants in the POME final discharge. GC/MS analyses also successfully identified cyclic volatile methyl siloxanes; cyclotetrasiloxane, octamethyl- (D4), cyclopentasiloxane, decamethyl- (D5), cyclohexasiloxane, dodecamethyl- (D6). D4 was detected at 0.0148-0.0357 mg/L, which could be potentially toxic. The palm oil industry used only water in the form of steam to process the fruits, and the presence of these compounds might be derived from the detergents and grease used in palm oil mill cleaning and maintenance operations. An appropriate treatment process is thus required to eliminate these toxicants from the POME final discharge. It is recommended that two approaches, chemical-based monitoring as well as biological toxicity-based monitoring, should be utilized for achieving an acceptable quality of POME final discharge in the future.

Pseudogene product YqiG is important for pflB expression and biohydrogen production in Escherichia coli BW25113.

Pseudogenes in the Escherichia coli genome are assumed to be non-functional. In this study, Keio collection BW25113∆yqiG and YqiG-producing strain (BW25113/pCA24N-YqiG) were used to evaluate the importance of pseudogene yqiG in hydrogen metabolism. Our results show pseudogene protein YqiG was identified as an essential protein in the production of biohydrogen from glucose. The mutant yqiG decreased biohydrogen production from 37 µmol mg-1 protein to 6 µmol mg-1 protein compared to the wild-type strain, and glucose consumption was reduced by 80%. Through transcriptional analysis, we found that the yqiG mutation represses pflB transcription tenfold; pflB encodes pyruvate-formate lyase, one of the key enzymes in the anaerobic metabolism of E. coli. Moreover, production of YqiG stimulated glycolysis and increased biohydrogen productivity 1.5-fold compared to that of the wild-type strain. Thus, YqiG is important for the central glycolysis reaction and is able to influence hydrogen metabolism activity in E. coli.

Subcritical Water-Carbon Dioxide Pretreatment of Oil Palm Mesocarp Fiber for Xylooligosaccharide and Glucose Production.

The present work aimed to investigate the pretreatment of oil palm mesocarp fiber (OPMF) in subcritical H2O-CO2 at a temperature range from 150⁻200 °C and 20⁻180 min with CO2 pressure from 3⁻5 MPa. The pretreated solids and liquids from this process were separated by filtration and characterized. Xylooligosaccharides (XOs), sugar monomers, acids, furans and phenols in the pretreated liquids were analyzed by using HPLC. XOs with a degree of polymerization X2⁻X4 comprising xylobiose, xylotriose, xylotetraose were analyzed by using HPAEC-PAD. Enzymatic hydrolysis was performed on cellulose-rich pretreated solids to observe xylose and glucose production. An optimal condition for XOs production was achieved at 180 °C, 60 min, 3 MPa and the highest XOs obtained was 81.60 mg/g which corresponded to 36.59% of XOs yield from total xylan of OPMF. The highest xylose and glucose yields obtained from pretreated solids were 29.96% and 84.65%, respectively at cellulase loading of 10 FPU/g-substrate.

Pre-treatment of Oil Palm Biomass for Fermentable Sugars Production.

Malaysia is the second largest palm oil producer in the world and this industry generates more than 80 million tonnes of biomass every year. When considering the potential of this biomass to be used as a fermentation feedstock, many studies have been conducted to develop a complete process for sugar production. One of the essential processes is the pre-treatment to modify the lignocellulosic components by altering the structural arrangement and/or removing lignin component to expose the internal structure of cellulose and hemicellulose for cellulases to digest it into sugars. Each of the pre-treatment processes that were developed has their own advantages and disadvantages, which are reviewed in this study.

Combination of Superheated Steam with Laccase Pretreatment Together with Size Reduction to Enhance Enzymatic Hydrolysis of Oil Palm Biomass.

The combination of superheated steam (SHS) with ligninolytic enzyme laccase pretreatment together with size reduction was conducted in order to enhance the enzymatic hydrolysis of oil palm biomass into glucose. The oil palm empty fruit bunch (OPEFB) and oil palm mesocarp fiber (OPMF) were pretreated with SHS and ground using a hammer mill to sizes of 2, 1, 0.5 and 0.25 mm before pretreatment using laccase to remove lignin. This study showed that reduction of size from raw to 0.25 mm plays important role in lignin degradation by laccase that removed 38.7% and 39.6% of the lignin from OPEFB and OPMF, respectively. The subsequent saccharification process of these pretreated OPEFB and OPMF generates glucose yields of 71.5% and 63.0%, which represent a 4.6 and 4.8-fold increase, respectively, as compared to untreated samples. This study showed that the combination of SHS with laccase pretreatment together with size reduction could enhance the glucose yield.

Soluble inhibitors generated during hydrothermal pretreatment of oil palm mesocarp fiber suppressed the catalytic activity of Acremonium cellulase.

Oil palm mesocarp fiber was subjected to hydrothermal pretreatment under isothermal and non-isothermal conditions. The pretreated slurries were separated by filtration, pretreated liquids and solids were characterized. An enzymatic digestibility study was performed for both pretreated slurries and solids to understand the effect of soluble inhibitors generated during the pretreatment process. The highest glucose yield obtained from pretreated slurries was 70.1%, and gradually decreased with higher pretreatment severities. The highest glucose yield obtained in pretreated solids was 100%, after pretreatment at 210°C for 20min. In order to study the inhibitory effects of compounds generated during pretreatment with cellulase, technical grade solutions that mimic the pretreated liquid were prepared and their effect on Acremonium cellulase activity was monitored using Avicel. Xylo-oligomers and tannic acid were identified as powerful inhibitors of Acremonium cellulase, and the lowest hydrolysis rate of Avicel of 0.18g/g-glucose released/L/h was obtained from tannic acid.

Combined pretreatment with hot compressed water and wet disk milling opened up oil palm biomass structure resulting in enhanced enzymatic digestibility.

Combined pretreatment with hot compressed water and wet disk milling was performed with the aim to reduce the natural recalcitrance of oil palm biomass by opening its structure and provide maximal access to cellulase attack. Oil palm empty fruit bunch and oil palm frond fiber were first hydrothermally pretreated at 150-190° C and 10-240 min. Further treatment with wet disk milling resulted in nanofibrillation of fiber which caused the loosening of the tight biomass structure, thus increasing the subsequent enzymatic conversion of cellulose to glucose. The effectiveness of the combined pretreatments was evaluated by chemical composition changes, power consumption, morphological alterations by SEM and the enzymatic digestibility of treated samples. At optimal pretreatment process, approximately 88.5% and 100.0% of total sugar yields were obtained from oil palm empty fruit bunch and oil palm frond fiber samples, which only consumed about 15.1 and 23.5 MJ/kg of biomass, respectively.

Hydrothermal and wet disk milling pretreatment for high conversion of biosugars from oil palm mesocarp fiber.

Eco-friendly pretreatment methods for lignocellulosic biomass are being developed as alternatives to chemical based methods. Superheated steam (SHS), hot compressed water (HCW) and wet disk milling (WDM) were used individually and with combination to partially remove hemicellulose and alter the lignin composition of recalcitrant structure of oil palm mesocarp fiber (OPMF). The efficiency of the pretreatment methods was evaluated based on the chemical compositions altered, SEM analysis, power consumption and degree of enzymatic digestibility. Hemicellulose removal (94.8%) was more pronounced under HCW compared to SHS, due to maximal contact of water and production of acetic acid which enhanced further degradation of hemicellulose. Subsequent treatment with WDM resulted in defibrillation of OPMF and expansion of the specific surface area thus increasing the conversion of cellulose to glucose. The highest glucose yield was 98.1% (g/g-substrate) when pretreated with HCW (200 °C, 20 min) and WDM which only consumed 9.6 MJ/kg of OPMF.

Hydrothermal pretreatment enhanced enzymatic hydrolysis and glucose production from oil palm biomass.

The present works investigate hydrothermal pretreatment of oil palm empty fruit bunch and oil palm frond fiber in a batch tube reactor system with temperature and time range from 170 to 250°C and 10 to 20min, respectively. The behavior of soluble sugars, acids, furans, and phenols dramatically changed over treatment severities as determined by HPLC. The cellulose-rich treated solids were analyzed by SEM, WAXD, and BET surface area. Enzymatic hydrolysis was performed from both pretreated slurries and washed solid, and data obtained suggested that tannic acid derived from lignin degradation was a potential cellulase inhibitor. Both partial removal of hemicellulose and migration of lignin during hydrothermal pretreatment caused structural changes on the cellulose-hemicellulose-lignin matrix, resulting in the opening and expansion of specific surface area and pore volume. The current results provided important factors that maximize conversion of cellulose to glucose from oil palm biomass by hydrothermal process.

Combined pretreatment using alkaline hydrothermal and ball milling to enhance enzymatic hydrolysis of oil palm mesocarp fiber.

Hydrothermal pretreatment of oil palm mesocarp fiber was conducted in tube reactor at treatment severity ranges of log Ro = 3.66-4.83 and partial removal of hemicellulose with migration of lignin was obtained. Concerning maximal recovery of glucose and xylose, 1.5% NaOH was impregnated in the system and subsequent ball milling treatment was employed to improve the conversion yield. The effects of combined hydrothermal and ball milling pretreatments were evaluated by chemical composition changes by using FT-IR, WAXD and morphological alterations by SEM. The successful of pretreatments were assessed by the degree of enzymatic digestibility of treated samples. The highest xylose and glucose yields obtained were 63.2% and 97.3% respectively at cellulase loadings of 10 FPU/g-substrate which is the highest conversion from OPMF ever reported.

Ball milling pretreatment of oil palm biomass for enhancing enzymatic hydrolysis.

Oil palm biomass, namely empty fruit bunch and frond fiber, were pretreated using a planetary ball mill. Particle sizes and crystallinity index values of the oil palm biomass were significantly reduced with extended ball mill processing time. The treatment efficiency was evaluated by the generation of glucose, xylose, and total sugar conversion yields from the pretreatment process compared to the amount of sugars from raw materials. Glucose and xylose contents were determined using high-performance liquid chromatography. An increasing trend in glucose and xylose yield as well as total sugar conversion yield was observed with decreasing particle size and crystallinity index. Oil palm frond fiber exhibited the best material yields using ball milling pretreatment with generated glucose, xylose, and total sugar conversion yields of 87.0, 81.6, and 85.4%, respectively. In contrast, oil palm empty fruit bunch afforded glucose and xylose of 70.0 and 82.3%, respectively. The results obtained in this study showed that ball mill-treated oil palm biomass is a suitable pretreatment method for high conversion of glucose and xylose.

Improved properties of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) produced by Comamonas sp. EB172 utilizing volatile fatty acids by regulating the nitrogen source.

This study presents the effect of carbon to nitrogen ratio (C/N) (mol/mol) on the cell growth and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) accumulation by Comamonas sp. EB172 in 2 L fermenters using volatile fatty acids (VFA) as the carbon source. This VFA was supplemented with ammonium sulphate and yeast extract in the feeding solution to achieve C/N (mol/mol) 5, 15, 25, and 34.4, respectively. By extrapolating the C/N and the source of nitrogen, the properties of the polymers can be regulated. The number average molecular weight (M n ) of P(3HB-co-3HV) copolymer reached the highest at 838 × 10(3) Da with polydispersity index (PDI) value of 1.8, when the culture broth was supplemented with yeast extract (C/N 34.4). Tensile strength and Young's modulus of the copolymer containing 6-8 mol% 3HV were in the ranges of 13-14.4 MPa and 0.26-0.34 GPa, respectively, comparable to those of polyethylene (PE). Thus, Comamonas sp. EB172 has shown promising bacterial isolates producing polyhydroxyalkanoates from renewable carbon materials.

Renewable sugars from oil palm frond juice as an alternative novel fermentation feedstock for value-added products.

In this paper, we report that pressed juice from oil palm frond (OPF) contained renewable sugars such as glucose, sucrose and fructose. By using a simple sugarcane press, 50% (wt/wt) of OPF juice was obtained from fresh OPF. The glucose content in the juice was 53.95±2.86g/l, which accounts for 70% of the total free sugars. We have examined the effect of various OPF juice concentrations on the production of poly(3-hydroxybutyrate), P(3HB) by Cupriavidus necator CCUG 52238(T). The cell dry mass in shake flask experiment reached 8.42g/l, with 32wt.% of P(3HB) at 30% (v/v) of OPF juice, comparable with using technical grade sugars. The biopolymer had a molecular mass, M(w) of 812kDa, with a low polydispersity index of 1.61. This result indicates that OPF juice can be used as an alternative renewable carbon source for P(3HB) production and has potential as a renewable carbon source.

PCR-based DGGE and FISH analysis of methanogens in an anaerobic closed digester tank for treating palm oil mill effluent

16S ribosomal RNA (rRNA)-targeted fluorescent in situ hybridization combined with polymerase chain reaction (PCR)-cloning, light microscopy using Gram stains, scanning electron microscopy and denatured gradient gel electrophoresis were used to reveal the distribution of methanogens within an anaerobic closed digester tank fed with palm oil mill effluent. For specific detection of methanogens, 16S rRNA-cloning analysis was conducted followed by restriction fragment length polymorphism (RFLP) for presumptive identification of methanogens. To cover the drawbacks of the PCR-cloning study, the organization of the microorganisms was visualized in the activated sludge sample by using fluorescent oligonucleotide probes specific to several different methanogens, and a probe for bacteria. In situ hybridization with methanogens and bacterial probes and denatured gradient gel electrophoresis within activated sludge clearly confirmed the presence of Methanosaeta sp. and Methanosarcina sp. cells. Methanosaeta concilii was found to be the dominant species in the bioreactor. These results revealed the presence of possibly new strain of Methanosaeta in the bioreactor for treating palm oil mill effluent called Methanosaeta concilii SamaliEB (Gene bank accession number: EU580025). In addition, fluorescent hybridization pictured the close association between the methanogens and bacteria and that the number of methanogens was greater than the number of bacteria.