Effective optimization of microalgae-to-bioethanol process systems hinges on an in-depth characterization of key process parameters relevant to the overall bioprocess engineering. One of the such important variables is the biomass particle size distribution and the effects on saccharification levels and bioethanol titres. This study examined the effects of three different microalgal biomass particle size ranges, 35 μm ≤ x ≤ 90 μm, 125 μm ≤ x ≤ 180 μm, and 295 μm ≤ x ≤ 425 μm, on the degree of enzymatic hydrolysis and bioethanol production. Two scenarios were investigated: single enzyme hydrolysis (cellulase) and double enzyme hydrolysis (cellulase and cellobiase). The glucose yield from biomass in the smallest particle size range (35 μm ≤ x ≤ 90 μm) was the highest, 134.73 mg glucose/g algae, while the yield from biomass in the larger particle size range (295 μm ≤ x ≤ 425 μm) was 75.45 mg glucose/g algae. A similar trend was observed for bioethanol yield, with the highest yield of 0.47 g EtOH/g glucose obtained from biomass in the smallest particle size range. The results have shown that the microalgal biomass particle size has a significant effect on enzymatic hydrolysis and bioethanol yield.
Bioprotein is one of the useful products obtained from biotechnology invention. It is a promising replacement for the commercial fish feed supplement. In this study, the enrichment of the bioprotein content after solid state fermentation using palm kernel cake and seaweed by the white rot fungus: Phanerochaete chrysoporium and yeast: Candida utilis was carried out. The growth media components were selected from 11 types of media using Plackett-Burman design (hereinafter PBD) and were optimized by one-factor-at-a-time (OFAT) method with bioprotein concentration (mg/g) as the response. From the screening result using PBD, three media components, namely K2HPO4, CuSO4.5H2O and MnSO4.H2O were selected for further optimization using OFAT method because of their positive contributions to the response. The final results showed that 5.0 g/L K2HPO4, 3.0 g/L CuSO4.5H2O and 0.1 g/L MnSO4.H2O were there to be the optimum media constituents with 9.0 g/L, MgSO4.7H2O, 0.1 g/L, CaCl2.H2O, 3.0 g/L FeSO4.7H2O and 3.0 g/L peptone as fixed compositions. At this optimum concentration, the protein increment of 11% was observed as compared to the results determined in the screening using PBD. The study revealed the benefits of using mixed cultures in improving the protein concentrations which can be used as nutritious fish feed.
Immobilized Candida antarctica lipase-catalyzed esterification of adipic acid and oleyl alcohol was investigated in a solvent-free system (SFS). Optimum conditions for adipate ester synthesis in a stirred-tank reactor were determined by the response surface methodology (RSM) approach with respect to important reaction parameters including time, temperature, agitation speed, and amount of enzyme. A high conversion yield was achieved using low enzyme amounts of 2.5% w/w at 60 degrees C, reaction time of 438 min, and agitation speed of 500 rpm. The good correlation between predicted value (96.0%) and actual value (95.5%) implies that the model derived from RSM allows better understanding of the effect of important reaction parameters on the lipase-catalyzed synthesis of adipate ester in an organic solvent-free system. Higher volumetric productivity compared to a solvent-based system was also offered by SFS. The results demonstrate that the solvent-free system is efficient for enzymatic synthesis of adipate ester.
In this study, the methanolysis process of sunflower oil was investigated to get high methyl esters (biodiesel) content using sodium methoxide. To reach to the best process conditions, central composite design (CCD) through response surface methodology (RSM) was employed. The optimal conditions predicted were the reaction time of 60 min, an excess stoichiometric amount of alcohol to oil ratio of 25%w/w and the catalyst content of 0.5%w/w, which lead to the highest methyl ester content (100%w/w). The methyl ester content of the mixture from gas chromatography analysis (GC) was compared to that of optimum point. Results, confirmed that there was no significant difference between the fatty acid methyl ester content of sunflower oil produced under the optimized condition and the experimental value (P ≥ 0.05). Furthermore, some fuel specifications of the resultant biodiesel were tested according to American standards for testing of materials (ASTM) methods. The outcome showed that the methyl ester mixture produced from the optimized condition met nearly most of the important biodiesel specifications recommended in ASTM D 6751 requirements. Thus, the sunflower oil methyl esters resulted from this study could be a suitable alternative for petrol diesels.
This study was undertaken to optimize skim milk and yeast extract concentration as a cultivation medium for optimal Bifidobacteria pseudocatenulatum G4 (G4) biomass and β -galactosidase production as well as lactose and free amino nitrogen (FAN) balance after cultivation period. Optimization process in this study involved four steps: screening for significant factors using 2(3) full factorial design, steepest ascent, optimization using FCCD-RSM, and verification. From screening steps, skim milk and yeast extract showed significant influence on the biomass production and, based on the steepest ascent step, middle points of skim milk (6% wt/vol) and yeast extract (1.89% wt/vol) were obtained. A polynomial regression model in FCCD-RSM revealed that both factors were found significant and the strongest influence was given by skim milk concentration. Optimum concentrations of skim milk and yeast extract for maximum biomass G4 and β -galactosidase production meanwhile low in lactose and FAN balance after cultivation period were 5.89% (wt/vol) and 2.31% (wt/vol), respectively. The validation experiments showed that the predicted and experimental values are not significantly different, indicating that the FCCD-RSM model developed is sufficient to describe the cultivation process of G4 using skim-milk-based medium with the addition of yeast extract.
The production of lignin from empty fruit bunch (EFB) has been carried out using liquefaction method with 1-butyl-3-methylimidazolium chloride ([BMIM]Cl) ionic liquid (IL), in presence of sulfuric acid (H2SO4) as a catalyst. Response surface methodology (RSM) based on a factorial Central Composite Design (CCD) was employed to identify the optimum condition for lignin yield. The result indicated that the second order model was adequate for all the independent variables on the response with R(2)=0.8609. The optimum temperature, time, ionic liquid to EFB ratio, and catalyst concentration were 150.5 °C, 151 min, 3:1 wt/wt and 4.73 wt%, respectively for lignin yield=26.6%. The presence of lignin liquefied product was confirmed by UV-Vis and FTIR analysis. It was also demonstrated lignin extraction from lignocellulosic using recycled IL gave sufficient performance.
Conversion of glucose, empty fruit bunch (efb) and kenaf to levulinic acid over a new hybrid catalyst has been investigated in this study. The characterization and catalytic performance results revealed that the physico-chemical properties of the new hybrid catalyst comprised of chromium chloride and HY zeolite increased the levulinic acid production from glucose compared to the parent catalysts. Optimization of the glucose conversion process using two level full factorial designs (2(3)) with two center points reported 55.2% of levulinic acid yield at 145.2 °C, 146.7 min and 12.0% of reaction temperature, reaction time and catalyst loading, respectively. Subsequently, the potential of efb and kenaf for producing levulinic acid at the optimum conditions was established after 53.2% and 66.1% of efficiencies were reported. The observation suggests that the hybrid catalyst has a potential to be used in biomass conversion to levulinic acid.
In this study optimization of drying oil palm trunk core lumber (OPTCL) biomass using microwave radiation was reported. Optimizing of the drying conditions using microwave, avoid burning, shrinkage and increasing the permeability of OPT was aimed to develop a new value added material. A set of experiments was designed by central composite design using response surface methodology (RSM) to statistically evaluate the findings. Three independent process variables including time (2-10 min), sample weight (300-1000 g) and input power (660-3300 W) were studied under the given conditions designed by Design Expert software. The results showed the effectiveness of microwave drying in reducing the time and better removal of moisture as compared to that of oven drying with no significant changes. Employing optimum conditions at 6.89 min of time with a microwave power set at 4 for a sample of 1000 g, predicting 14.62% of moisture content.
Induction strategies for the periplasmic production of recombinant human IFN-alpha2b (interferon-alpha2b) by recombinant Escherichia coli Rosetta-gami 2(DE3) were optimized in shake-flask cultures using response surface methodology based on the central composite design. The factors included in the present study were induction point, which related to the attenuance of the cell culture, IPTG (isopropyl beta-D-thiogalactoside) concentration and induction temperature. Second-order polynomial models were used to correlate the abovementioned factors to soluble periplasmic IFN-alpha2b formation and percentage of soluble IFN-alpha2b translocated to the periplasmic space of E. coli. The models were found to be significant and subsequently validated. The proposed induction strategies consisted of induction at an attenuance of 4 (measured as D600), IPTG concentration of 0.05 mM and temperature of 25 degrees C. The optimized induction strategy reduced inclusion-body formation as evidenced by electron microscopy and yielded 323.8 ng/ml of IFN-alpha2b in the periplasmic space with translocation of 74% of the total soluble product. In comparison with the non-optimized condition, soluble periplasmic production and the percentage of soluble IFN-alpha2b translocated to the periplasmic space obtained in optimized induction strategies were increased by approx. 20-fold and 1.4-fold respectively.
Pretreatment of the high free fatty acid rubber seed oil (RSO) via esterification reaction has been investigated by using a pilot scale hydrodynamic cavitation (HC) reactor. Four newly designed orifice plate geometries are studied. Cavities are induced by assisted double diaphragm pump in the range of 1-3.5 bar inlet pressure. An optimised plate with 21 holes of 1mm diameter and inlet pressure of 3 bar resulted in RSO acid value reduction from 72.36 to 2.64 mg KOH/g within 30 min of reaction time. Reaction parameters have been optimised by using response surface methodology and found as methanol to oil ratio of 6:1, catalyst concentration of 8 wt%, reaction time of 30 min and reaction temperature of 55°C. The reaction time and esterified efficiency of HC was three fold shorter and four fold higher than mechanical stirring. This makes the HC process more environmental friendly.
To explore new approaches of phage-based bio-process of specifically pathogenic Escherichia coli bacteria in food products within a short period. One hundred and forty highly lytic designed coliphages were used. Escherichia coli naturally contaminated and Enterohemorrhagic Escherichia coli experimentally inoculated samples of lettuce, cabbage, meat, and egg were used. In addition, experimentally produced biofilms of E. coli were tested. A phage concentration of 10(3) PFU/ml was used for food products immersion, and for spraying of food products, 10(5) PFU/ml of a phage cocktail was used by applying a 20-s optimal dipping time in a phage cocktail. Food samples were cut into pieces and were either sprayed with or held in a bag immersed in lambda buffer containing a cocktail of 140 phages. Phage bio-processing was successful in eliminating completely E. coli in all processed samples after 48 h storage at 4°C. Partial elimination of E. coli was observed in earlier storage periods (7 and 18 h) at 24° and 37°C. Moreover, E. coli biofilms were reduced >3 log cycles upon using the current phage bio-processing. The use of a phage cocktail of 140 highly lytic designed phages proved highly effective in suppressing E. coli contaminating food products. Proper decontamination/prevention methods of pathogenic E. coli achieved in this study can replace the current chemically less effective decontamination methods.
Microalgae have caught the world's attention for its potential to solve one of the world's most pressing issues-sustainable green energy. Compared to biofuels supplied by oil palm, rapeseed, soybean and sugar cane, microalgae alone can be manipulated to generate larger amounts of biodiesel, bioethanol, biohydrogen and biomass in a shorter time. Apart from higher productivity, microalgae can also grow using brackish water on non-arable land, greatly reducing the competition with food and cash crops. Hence, numerous efforts have been put into the commercialisation of microalgae-derived biofuel by both the government and private bodies. This paper serves to review conventional and novel methods for microalgae culture and biomass harvest, as well as recent developments in techniques for microalgal biofuel production.
The nanobiocatalyst (NBC) is an emerging innovation that synergistically integrates advanced nanotechnology with biotechnology and promises exciting advantages for improving enzyme activity, stability, capability and engineering performances in bioprocessing applications. NBCs are fabricated by immobilizing enzymes with functional nanomaterials as enzyme carriers or containers. In this paper, we review the recent developments of novel nanocarriers/nanocontainers with advanced hierarchical porous structures for retaining enzymes, such as nanofibres (NFs), mesoporous nanocarriers and nanocages. Strategies for immobilizing enzymes onto nanocarriers made from polymers, silicas, carbons and metals by physical adsorption, covalent binding, cross-linking or specific ligand spacers are discussed. The resulting NBCs are critically evaluated in terms of their bioprocessing performances. Excellent performances are demonstrated through enhanced NBC catalytic activity and stability due to conformational changes upon immobilization and localized nanoenvironments, and NBC reutilization by assembling magnetic nanoparticles into NBCs to defray the high operational costs associated with enzyme production and nanocarrier synthesis. We also highlight several challenges associated with the NBC-driven bioprocess applications, including the maturation of large-scale nanocarrier synthesis, design and development of bioreactors to accommodate NBCs, and long-term operations of NBCs. We suggest these challenges are to be addressed through joint collaboration of chemists, engineers and material scientists. Finally, we have demonstrated the great potential of NBCs in manufacturing bioprocesses in the near future through successful laboratory trials of NBCs in carbohydrate hydrolysis, biofuel production and biotransformation.
The attention on genetically modified (GM) food industry is increasing due to the flourishing
of biotechnology. However, there are some debates on the associated benefits and risks of
employing modification technology in food industry. This study strives to examine the causes
that determine consumers’ benefit and risk perceptions on GM foods. Besides, the influence of
perceived benefit and risk of GM food on consumers’ attitude is investigated. The empirical
results of this study showed that GM food knowledge, and GM food characteristics have been
acting as important predictors of both benefits and risks perceptions. Further, it is also found
that perceived benefits showed significant positive influence on attitude, and attitude affects
purchase intention towards GM food. Research implications to policy makers, scientists, and
market practitioners are covered, in which suggestions and recommendations are provided
to these parties. Lastly, research implications and recommendations to future research are
discussed.
This preliminary study aimed to identify a commercial gamat species, Stichopus horrens Selenka, 1867, and a timun laut species, Holothuria (Mertensiothuria) leucospilota (Brandt, 1835), from Pangkor Island, Perak, Malaysia, employing morphological techniques based on the shape of the ossicles and molecular techniques based on the cytochrome c oxidase I (COI) mitochondrial DNA (mtDNA) gene. In Malaysia, a gamat is defined as a sea cucumber species of the family Stichopodidae with medicinal value, and timun laut refers to non-gamat species. S. horrens is very popular on Pangkor Island as a main ingredient in the traditional production of air gamat and minyak gamat, while H. leucospilota is the most abundant species in Malaysia. In contrast to previous studies, internal body parts (the respiratory tree and gastrointestine) were examined in this study to obtain better inferences based on morphology. The results showed that there were no ossicles present in the gastrointestine of H. leucospilota, and this characteristic is suggested as a unique diagnostic marker for the timun laut species. In addition, the presence of Y-shaped rods in the respiratory tree of S. horrens subsequently supported the potential to use internal body parts to identify the gamat species. Phylogenetic analysis of the COI mtDNA gene of the sea cucumber specimens using the neighbour-joining method and maximum likelihood methods further confirmed the species status of H. leucospilota and S. horrens from Pangkor Island, Perak, Malaysia. The COI mtDNA gene sequences were registered with GenBank, National Center for Biotechnology Information (NCBI), US National Library of Medicine (GenBank accession no.: KC405565-KC405568). Although additional specimens from various localities will be required to produce more conclusive results, the current findings provide better insight into the importance of complementary approaches involving morphological and molecular techniques in the identification of the two Malaysian sea cucumber species.
The simultaneous production of hydrogen and ethanol by microorganisms from waste materials in a bioreactor system would establish cost-effective and time-saving biofuel production. This review aims to present the current status of fermentation processes producing hydrogen accompanied by ethanol as a co-product. We outlined the microbes used and their fundamental pathways for hydrogen and ethanol fermentation. Moreover, we discussed the exploitation of renewable and sustainable waste materials as promising feedstock and the limitations encountered. The low substrate bioconversion rate in hydrogen and ethanol co-production is regarded as the primary constraint towards the development of large scale applications. Thus, microbes with an enhanced capability have been generated via genetic manipulation to diminish the inefficiency of substrate consumption. In this review, other potential approaches to improve the performance of co-production through fermentation were also elaborated. This review will be a useful guide for the future development of hydrogen and ethanol co-production using waste materials.
Preparation of activated carbon has been attempted using KOH as activating agent by microwave heating from biodiesel industry solid residue, oil palm empty fruit bunch (EFBAC). The significance of chemical impregnation ratio (IR), microwave power and activation time on the properties of activated carbon were investigated. The optimum condition has been identified at the IR of 1.0, microwave power of 600 W and activation time of 7 min. EFBAC was characterized by scanning electron microscopy, Fourier transform infrared spectroscopy and nitrogen adsorption isotherm. The surface chemistry was examined by zeta potential measurement, determination of surface acidity/basicity, while the adsorptive property was quantified using methylene blue as dye model compound. The optimum conditions resulted in activated carbon with a monolayer adsorption capacity of 395.30 mg/g and carbon yield of 73.78%, while the BET surface area and total pore volume were corresponding to 1372 m2/g and 0.76 cm3/g, respectively.
The synthesis of fatty acid ethyl esters (FAEEs) by a two-step in situ (reactive) esterification/transesterification from Jatropha curcas L. (JCL) seeds using microwave system has been investigated. Free fatty acid was reduced from 14% to less than 1% in the first step using H2SO4 as acid catalyst after 35 min of microwave irradiation heating. The organic phase in the first step was subjected to a second reaction by adding 5 N KOH in ethanol as the basic catalyst. Response surface methodology (RSM) based on central composite design (CCD) was utilized to design the experiments and analyze the influence of process variables (particles seed size, time of irradiation, agitation speed and catalyst loading) on conversion of triglycerides (TGs) in the second step. The highest triglycerides conversion to fatty acid ethyl esters (FAEEs) was 97.29% at the optimum conditions:<0.5mm seed size, 12.21 min irradiation time, 8.15 ml KOH catalyst loading and 331.52 rpm agitation speed in the 110 W microwave power system.
Microalgal-bacterial granular sludge (MBGS) process has become a focal point in treating municipal wastewater. However, it remains elusive whether the emerging process can be applied for the treatment of aquaculture wastewater, which contains considerable concentrations of nitrate and nitrite. This study evaluated the feasibility of MBGS process for aquaculture wastewater treatment. Result showed that the MBGS process was competent to remove respective 64.8%, 84.9%, 70.8%, 50.0% and 84.2% of chemical oxygen demand, ammonia-nitrogen, nitrate-nitrogen, nitrite-nitrogen and phosphate-phosphorus under non-aerated conditions within 8 h. The dominant microalgae and bacteria were identified to be Coelastrella and Rhodobacteraceae, respectively. Further metagenomics analysis implied that microbial assimilation was the main contributor in organics, nitrogen and phosphorus removal. Specifically, considerable nitrate and nitrite removals were also obtained with the synergy between microalgae and bacteria. Consequently, this work demonstrated that the MBGS process showed a prospect of becoming an environmentally friendly and efficient alternative in aquaculture wastewater treatment.