Optimization of Saccharification and Fermentation Process in Bioethanol Production from Oil Palm Fronds

Oil Palm Frond (OPF) is one of lignocellulosic biomass, which can be utilized as raw material for bioethanol production. Bioethanol is produced as alternative energy to substitute gasoline. There are four steps in bioethanol production from OPF, i.e pretreatement, saccharification, fermentation and purification process. In this study, optimization of saccharification and fermentation process for OPF was investigated. Two methods and the variations of enzyme concentration were carried out in the saccharification and fermentation process. Separate hydrolysis and fermentation process (SHF) and simultaneous saccharification and fermentation process (SSF) were conducted to produce ethanol optimally. Variations of enzyme concentration used in this process were 10, 20, 30 and 40 FPU/g substrate. The result shows that the highest ethanol concentration can be obtained in SSF process with 30 FPU/g substrate of enzyme concentration. The process produced 59.20 g/L ethanol (95.95% yield ethanol) at 96 h of SSF process.     

Industrial Waste Utilization for Low-Cost Production of Raw Material Oil Through Microbial Fermentation

In view of ever-growing demand of biodiesel, there is an urgent need to look for inexpensive and promising renewable raw material oils for its production. In this context, the aim of this study was to evaluate the potential use of industrial wastes for low-cost production of oils through microbial fermentation. Among the strains tested, Yarrowia lipolytica grew best and produced highest lipid when grown on decanter effluent from palm oil mill. When crude glycerol by-product from a biodiesel plant was added into the effluent as a co-substrate, Y. lipolytica produced a higher biomass of 3.21 g/L and a higher amount of lipid of 2.21 g/L which was 68 % of the dry weight. The scale up and process improvement in a 5-L bioreactor increased the biomass and lipid up to 5.53 and 2.81 g/L, respectively. A semi-continuous mode of operation was an effective mode for biomass enhancement while a fed-batch mode was effective for lipid enhancement. These yeast lipids have potential to be used as biodiesel feedstocks because of their similar fatty acid composition to that of plant oil.     

Rhodococcus as Biofactories for Microbial Oil Production

Bacteria belonging to the Rhodococcus genus are frequent components of microbial communities in diverse natural environments. Some rhodococcal species exhibit the outstanding ability to produce significant amounts of triacylglycerols (TAG) (>20% of cellular dry weight) in the presence of an excess of the carbon source and limitation of the nitrogen source. For this reason, they can be considered as oleaginous microorganisms. As occurs as well in eukaryotic single-cell oil (SCO) producers, these bacteria possess specific physiological properties and molecular mechanisms that differentiate them from other microorganisms unable to synthesize TAG. In this review, we summarized several of the well-characterized molecular mechanisms that enable oleaginous rhodococci to produce significant amounts of SCO. Furthermore, we highlighted the ability of these microorganisms to degrade a wide range of carbon sources coupled to lipogenesis. The qualitative and quantitative oil production by rhodococci from diverse industrial wastes has also been included. Finally, we summarized the genetic and metabolic approaches applied to oleaginous rhodococci to improve SCO production. This review provides a comprehensive and integrating vision on the potential of oleaginous rhodococci to be considered as microbial biofactories for microbial oil production.     

Solid-State Fermentation for Humic Acids Production by a Trichoderma reesei Strain Using an Oil Palm Empty Fruit Bunch as the Substrate

Empty fruit bunch (EFB), an underutilized waste product of oil palm processing, was studied as a substrate for the production of humic acids (HA) by a Trichoderma reesei strain by solid-state fermentation (SSF) in Raimbault columns. HA have attracted the attention of many investigators due to their applications in agriculture, industry, the environment, and biomedicine. Commercial HA are currently chemically extracted from peat and coal, which are nonrenewable carbon sources. Biotechnological processes are important for their sustainable and controlled production, with SSF being especially promising for mimicking the natural habitat of fungi. Trichoderma sporulation and HA production are related, and the results of this study showed that SSF stimulated fast sporulation. The productivity related to HA was much higher than that of the biomass, indicating an efficient utilization of EFB. These findings, added to the low cost of EFB, make SSF an attractive process for HA production.     

Lipase-Catalyzed Synthesis of Cocoa Butter Equivalent from Palm Olein and Saturated Fatty Acid Distillate from Palm Oil Physical Refinery

Cocoa butter equivalent was prepared by enzymatic acidolysis reaction of substrate consisting of refined palm olein oil and palmitic?Cstearic fatty acid mixture. The reactions were performed in a batch reactor at a temperature of 60?°C in an orbital shaker operated at 160?RPM. Different mass ratios of substrates were explored and the compositions of the five major triacylglycerol (TAG) of the structured lipids were identified and quantified using cocoa butter-certified reference material IRMM-801. The reaction resulted in production of cococa butter equivent with TAG compostion (POP 26.6 %, POS 42.1, POO 7.5, SOS 18.0 %, and SOO 5.8 %) and melting temperature between 34.7 and 39.6?°C which is close to that of the cocoa butter. The result of this research demonstrated the potential use of saturated fatty acid distillate (palmitic and stearic fatty acids) obtained from palm oil physical refining process into a value-added product.     

Fed-batch Anaerobic Valorization of Slaughterhouse By-products with Mesophilic Microbial Consortia Without Methane Production

This work aimed at setting up a fully instrumented, laboratory-scale bioreactor enabling anaerobic valorization of solid substrates through hydrogen and/or volatile fatty acid (VFA) production using mixed microbial populations (consortia). The substrate used was made of meat-based wastes, especially from slaughterhouses, which are becoming available in large amounts as a consequence of the growing constraints for waste disposal from meat industry. A reconstituted microbial mesophilic consortium without Archaebacteria (methanogens), named PBr, was cultivated in a 5-L anaerobic bioreactor on slaughterhouse wastes. The experiments were carried out with sequential fed-batch operations, including liquid medium removal from the bioreactor and addition of fresh substrate. VFAs and nitrogen were the main metabolites observed, while hydrogen accumulation was very low and no methane production was evidenced. After 1,300 h of culture, yields obtained for VFAs reached 0.38 g/g dry matter. Strain composition of the microbial consortium was also characterized using molecular tools (temporal temperature gradient gel electrophoresis and gene sequencing).     

Valorization and Application of Fruit and Vegetable Wastes and By-Products for Food Packaging Materials

The important roles of food packaging are food protection and preservation during processing, transportation, and storage. Food can be altered biologically, chemically, and physically if the packaging is unsuitable or mechanically damaged. Furthermore, packaging is an important marketing and communication tool to consumers. Due to the worldwide problem of environmental pollution by microplastics and the large amounts of unused food wastes and by-products from the food industry, it is important to find more environmentally friendly alternatives. Edible and functional food packaging may be a suitable alternative to reduce food waste and avoid the use of non-degradable plastics. In the present review, the production and assessment of edible food packaging from food waste as well as fruit and vegetable by-products and their applications are demonstrated. Innovative food packaging made of biopolymers and biocomposites, as well as active packaging, intelligent packaging, edible films, and coatings are covered.     

Enzymatic Synthesis of Biodiesel via Alcoholysis of Palm Oil

The enzymatic alcoholysis of crude palm oil with methanol and ethanol was investigated using commercial immobilized lipases (Lipozyme RM IM, Lipozyme TL IM). The effect of alcohol (methanol or ethanol), molar ratio of alcohol to crude palm oil, and temperature on biodiesel production was determined. The best ethyl ester yield was about 25 wt.% and was obtained with ethanol/oil molar ratio of 3.0, temperature of 50 °C, enzyme concentration of 3.0 wt.%, and stepwise addition of the alcohol after 4 h of reaction. Experiments with 1 and 3 wt.% of KOH and 3 wt.% of MgO were carried out to compare their catalytic behavior with the enzymatic transesterification results. The commercial immobilized lipase, Lipozyme TL IM, showed the best catalytic performance.     

无溶剂体系中脂肪酶催化合成单甘酯的研究

以自制铜绿假单胞菌脂肪酶为催化剂,通过无溶剂法棕榈油甘油解反应催化合成了单脂肪酸甘油酯,实验结果表明：适宜加酶量为500u/g棕榈 油,甘油相适宜水含量为3％－4．5％,适宜加酶量为500u/g棕榈油,甘油与棕榈油的摩尔比为（2．0－2．5）：1,最佳反应温度为38－42度,在此温度区间内适宜反应时间为24h,临界反应温度为44度,程序降温和批次加酶更有利于反应,产物中单甘酯含量可增大10－20％,实验条件下反应48h后单甘酯含量可达65％,脱酶后达77％。     

Oil Crop Biomass Residue-Based Media for Enhanced Algal Lipid Production

The aim of this study was to evaluate the use of hydrolysates from acid hydrolysis of four different oil crop biomass residues (OCBR) as low cost culture media for algae growth. The one-factor-at-a-time method was used to design a series of experiments to optimize the acid hydrolysis conditions through examining the total nitrogen, total phosphorus, chemical oxygen demand, and ammonia nitrogen in the hydrolysates. The optimal conditions were found to be using 3 % sulfuric acid and hydrolyzing residues at 90 °C for 20 h. The hydrolysates (OCBR media) produced under the optimal conditions were used to cultivate the two algae strains, namely UM258 and UM268. The results from 5 days of cultivation showed that the OCBR media supported faster algae growth with maximal algal biomass yield of 2.7 and 3 g/L, respectively. Moreover, the total lipids for UM258 and UM268 were 54 and 35 %, respectively, after 5 days of cultivation, which suggested that the OCBR media allowed the algae strains to accumulate higher lipids probably due to high C/N ratio. Furthermore, over 3 % of omega-3 fatty acid (EPA) was produced for the two algae strains. In conclusion, OCBR media are excellent alternative for algae growth and have a great potential for large-scale production of algae-based ingredients for biodiesel as well as high-value food and pharmaceutical products.     

Valorization of a By-Product from the Production of Mechanically Deboned Chicken Meat for Preparation of Gelatins

In recent decades, food waste management has become a key priority of industrial and food companies, state authorities and consumers as well. The paper describes the biotechnological processing of mechanically deboned chicken meat (MDCM) by-product, rich in collagen, into gelatins. A factorial design at two levels was used to study three selected process conditions (enzyme conditioning time, gelatin extraction temperature and gelatin extraction time). The efficiency of the technological process of valorization of MDCM by-product into gelatins was evaluated by % conversion of the by-product into gelatins and some qualitative parameters of gelatins (gel strength, viscosity and ash content). Under optimal processing conditions (48–72 h of enzyme conditioning time, 73–78 °C gelatin extraction temperature and 100–150 min gelatin extraction time), MDCM by-product can be processed with 30–32% efficiency into gelatins with a gel strength of 140 Bloom, a viscosity of 2.5 mPa.s and an ash content of 5.0% (which can be reduced by deionization using ion-exchange resins). MDCM is a promising food by-product for valorization into gelatins, which have potential applications in food-, pharmaceutical- and cosmetic fields. The presented technology contributes not only to food sustainability but also to the model of a circular economy.     

Efficient Production of Ethanol from Empty Palm Fruit Bunch Fibers by Fed-Batch Simultaneous Saccharification and Fermentation Using Saccharomyces cerevisiae

The concentration of ethanol produced from lignocellulosic biomass should be at least 40 g l^?1 [about 5 % (v/v)] to minimize the cost of distillation process. In this study, the conditions for the simultaneous saccharification and fermentation (SSF) at fed-batch mode for the production of ethanol from alkali-pretreated empty palm fruit bunch fibers (EFB) were investigated. Optimal conditions for the production of ethanol were identified as temperature, 30 °C; enzyme loading, 15 filter paper unit g^?1 biomass; and yeast (Saccharomyces cerevisiae) loading, 5 g l^?1 of dry cell weight. Under these conditions, an economical ethanol concentration was achieved within 17 h, which further increased up to 62.5 g l^?1 after 95 h with 70.6 % of the theoretical yield. To our knowledge, this is the first report to evaluate the economic ethanol production from alkali-pretreated EFB in fed-batch SSF using S. cerevisiae.     

Biotechnological Production of Phenyllactic Acid and Biosurfactants from Trimming Vine Shoot Hydrolyzates by Microbial Coculture Fermentation

Coculture fermentations show advantages for producing food additives from agroindustrial wastes, considering that different specified microbial strains are combined to improve the consumption of mixed sugars obtained by hydrolysis. This technology dovetails with both the growing interest of consumers towards the use of natural food additives and with stricter legislations and concern in developed countries towards the management of wastes. The use of this technology allows valorization of both cellulosic and hemicellulosic fractions of trimming vine shoots for the production of lactic acid (LA), phenyllactic acid (PLA), and biosurfactants (BS). This work compares the study of the potential of hemicellulosic and cellulosic fractions of trimming vine shoots as cheaper and renewable carbon sources for PLA and BS production by independent or coculture fermentations. The highest LA and PLA concentrations, 43.0 g/L and 1.58 mM, respectively, were obtained after 144 h during the fermentation of hemicellulosic sugars and simultaneous saccharification and fermentation (SSF) carried out by cocultures of Lactobacillus plantarum and Lactobacillus pentosus. Additionally, cell-bond BS decreased the surface tension (ST) in 17.2 U; meanwhile, cell-free supernatants (CFS) showed antimicrobial activity against Salmonella enterica and Listeria monocytogenes with inhibition halos of 12.1?±?0.6 mm and 11.5?±?0.9 mm, respectively.     

高效液相色谱法定量测定采油微生物发酵菌液中的有机酸

采用反相高效液相色谱法(RP-HPLC)在Kromasil C18分析柱上,以0.015 mol/L磷酸二氢钠缓冲液(pH=2.3)和乙腈(体积比为96∶4)的二元流动相分离测定了采油微生物发酵液中的有机酸,流动相的流速为1.0mL/min,采用紫外检测器在210 nm处对有机酸进行测定.结果表明有机酸浓度在0.05～1.5 g/L时其峰面积与相应的质量浓度有良好的线性关系,其r大小等于0.9990,测定结果的RSD都小于4.0%(n=8),平均回收率均为100±4%以内.该方法具有测定结果准确可靠,操作简便快速,重现性好等特点.     

Kaolin-Carbon Adsorbents for Carotene Removal of Red Palm Oil

Kaolin-carbon adsorbents were prepared with and without sulfuric acid pretreatment followed by activation-carbonization at 500 degrees C. The total surface area of the resulting kaolin-carbon adsorbents was found to be decreased with the increase in kaolin loading. Sulfuric acid pretreatment of the precursor prior to the carbonization-activation processes resulted in the enhancement of total surface area but reduced the micropore surface area of the resulting adsorbents. At the same time, this improved carotene adsorption capacity from red palm oil. However, recovery of carotene from the carotene-adsorbed adsorbent is only improved when the acid pretreatment of the precursor was done at a high loading percentage of activated carbon. Similarly, the peroxide value (PV) increased. A maximum removal of carotene from red palm oil was obtained at 20% kaolin loading for both adsorbents prepared with and without sulfuric acid pretreatment with about 45 and 65% carotene removal, respectively, from a 30-ppm solution. This indicates that pretreatment with sulfuric acid, prior to the activation-carbonization process, increased the carotene uptake by the resulting adsorbent. However, a further increase in the kaolin loading resulted in the decrease of carotene removal. About 3-4% of carotene adsorbed can be recovered from both types of adsorbents under optimum condition, in which the percentage recovered decreased with the increase in kaolin loading. On the other hand, the PV increased with kaolin loading at around 54-64 mEq/kg for both types of adsorbents. It was also found that carotene uptake by the adsorbents is high if the adsorbent contains a high percentage of activated carbon. Similarly, carotene recovery is high and less oxidation can be observed, as indicated by the lower PV value. Copyright 2001 Academic Press.     

Production of Ethanol and Feed by High Dry Matter Hydrolysis and Fermentation of Palm Kernel Press Cake

Palm kernel press cake (PKC) is a residue from palm oil extraction presently only used as a low protein feed supplement. PKC contains 50% fermentable hexose sugars present in the form of glucan and mainly galactomannan. This makes PKC an interesting feedstock for processing into bioethanol or in other biorefinery processes. Using a combination of mannanase, β-mannosidase, and cellulases, it was possible without any pretreatment to hydrolyze PKC at solid concentrations of 35% dry matter with mannose yields up to 88% of theoretical. Fermentation was tested using Saccharomyces cerevisiae in both a separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) setup. The hydrolysates could readily be fermented without addition of nutrients and with average fermentation yields of 0.43?±?0.02 g/g based on consumed mannose and glucose. Employing SSF, final ethanol concentrations of 70 g/kg was achieved in 216 h, corresponding to an ethanol yield of 70% of theoretical or 200 g ethanol/kg PKC. Testing various enzyme mixtures revealed that including cellulases in combination with mannanases significantly improved ethanol yields. Processing PKC to ethanol resulted in a solid residue enriched in protein from 17% to 28%, a 70% increase, thereby potentially making a high-protein containing feed supplement.     

Crude Cellulase from Oil Palm Empty Fruit Bunch by Trichoderma asperellum UPM1 and Aspergillus fumigatus UPM2 for Fermentable Sugars Production

Cellulase is an enzyme that converts the polymer structure of polysaccharides into fermentable sugars. The high market demand for this enzyme together with the variety of applications in the industry has brought the research on cellulase into focus. In this study, crude cellulase was produced from oil palm empty fruit bunch (OPEFB) pretreated with 2 % NaOH with autoclave, which was composed of 59.7 % cellulose, 21.6 % hemicellulose, and 12.3 % lignin using Trichoderma asperellum UPM1 and Aspergillus fumigatus UPM2. Approximately 0.8 U/ml of FPase, 24.7 U/ml of CMCase and 5.0 U/ml of β-glucosidase were produced by T. asperellum UPM1 at a temperature of 35 °C and at an initial pH of 7.0. A 1.7 U/ml of FPase, 24.2 U/ml of CMCase, and 1.1 U/ml of β-glucosidase were produced by A. fumigatus UPM2 at a temperature of 45 °C and at initial pH of 6.0. The crude cellulase was best produced at 1 % of substrate concentration for both T. asperellum UPM1 and A. fumigatus UPM2. The hydrolysis percentage of pretreated OPEFB using 5 % of crude cellulase concentration from T. asperellum UPM1 and A. fumigatus UPM2 were 3.33 % and 19.11 %, with the reducing sugars concentration of 1.47 and 8.63 g/l, respectively.