Search results for: cellulose hydrolysis

Authors: Ridzuan Ramli, Zianor Azrina Zianon Abdin, Mohammad Dalour Beg, Rosli M. Yunus

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Nanocrystalline cellulose (NCC) were produced by using the ultrasound assisted acid hydrolysis from oil palm empty fruit bunch (EFB) pulp with different hydrolysis time then were analyzed by using FESEM and TGA as in comparison with EFB fiber and EFB pulp. Based on the FESEM analysis, it was found that NCC has a rod like shaped under the acid hydrolysis with an assistant of ultrasound. According to thermal stability, the NCC obtained show remarkable sign of high thermal stability compared to EFB fiber and EFB pulp. However, as the hydrolysis time increase, the thermal stability of NCC was deceased. As in conclusion, the NCC can be prepared by using ultrasound assisted acid hydrolysis. The NCC obtained have good thermal stability and have a great potential as the reinforcement in composite materials.

Keywords: Nanocrystalline cellulose, ultrasound assisted acid hydrolysis, thermal stability, morphology, empty fruit bunch (EFB)

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Authors: Akida Mulyaningtyas, Wahyudi Budi Sediawan

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One decisive stage in bioethanol production from plant biomass is the hydrolysis of lignocellulosic materials into simple sugars such as glucose. The produced glucose is then fermented into ethanol. This stage is popularly done in biological method by using cellulase that is produced by certain fungi. As it is known, glucose is the main source of nutrition for most microorganisms. Therefore, cutting cellulose into glucose is actually an attempt of microorganism to provide nutrition for itself. So far, this phenomenon has received less attention while it is necessary to identify the quantity of sugar consumed by the microorganism. In this study, we examined the phenomenon of sugar consumption by microorganism on lignocellulosic hydrolysis. We used oil palm empty fruit bunch (OPEFB) as the source of lignocellulose and Aspergillus niger as cellulase-producing fungus. In Indonesia, OPEFB is plantation waste that is difficult to decompose in nature and causes environmental problems. First, OPEFB was pretreated with 1% of NaOH at 170 oC to destroy lignin that hindered A.niger from accessing cellulose. The hydrolysis was performed by growing A.niger on pretreated OPEFB in solid state to minimize the possibility of contamination. The produced glucose was measured every 24 hours for 9 days. We analyzed the kinetics of both reactions, i.e., hydrolysis and glucose consumption, simultaneously. The constants for both reactions were assumed to follow the Monod equation. The results showed that the reaction constant of glucose consumption (μC) was higher than of cellulose hydrolysis (μH), i.e., 11.8 g/L and 0.62 g/L for glucose consumption and hydrolysis respectively. However, in general, the reaction rate of hydrolysis is greater than of glucose consumption since the cellulose concentration as substrate in hydrolysis is much higher than glucose as substrate in the consumption reaction.

Keywords: Aspergillus niger, bioethanol, hydrolysis, kinetics

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Authors: Ife O. Bolaji

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Environmental concerns arising from the use of fossil fuels has increased the interest in the development of renewable and sustainable sources of energy. This would minimize the dependence on fossil fuels and serve as future alternatives. Organic wastes contain carbohydrates, proteins and lipids, which can be utilised as carbon sources for the production of bio-based products. Cellulose is the most abundant natural biopolymer, being the main structural component of lignocellulosic materials. The aim of this project is to develop a biological process for the hydrolysis and fermentation of organic wastes into ethanol and organic acids. The hydrolysis and fermentation processes are integrated in a single vessel using undefined mixed culture microorganisms. The anaerobic fermentation of microcrystalline cellulose was investigated in continuous and batch reactors at 25°C with an appropriate growth medium for cellulase formation, hydrolysis, and fermentation. The reactors were inoculated with soil (B1, C1, C3) or sludge from an anaerobic digester (B2, C2) and the breakdown of cellulose was monitored by measuring the production of ethanol, organic acids and the residual cellulose. The batch reactors B1 and B2 showed negligible microbial activity due to inhibition while the continuous reactors, C1, C2 and C3, exhibited little cellulose hydrolysis which was concealed by the cellulose accumulation in the reactor. At the end of the continuous operation, the reactors C1, C2 and C3 were operated under batch conditions. 48%, 34% and 42% cellulose had been fermented by day 88, 55 and 55 respectively of the batch fermentation. Acetic acid, ethanol, propionic acid and butyric acids were the main fermentation products in the reactors. A stable concentration of 0.6 g/l ethanol and 5 g/L acetic acid was maintained in C3 for several weeks due to reduced activity of methanogens caused by the decrease in pH. Thus far, the results have demonstrated that mixed microbial culture is capable of hydrolysing and fermenting cellulose under lenient conditions. The fermentation of cellulose has been found effective in a combination of continuous and batch processes.

Keywords: cellulose, hydrolysis, mixed culture, organic waste

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Authors: Md. Sakinul Islam, Nhol Kao, Sati Bhattacharya, Rahul Gupta

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The objective of the study is to produce naocrystalline cellulose (NCC) from rice husk by chemical extraction process. The chemical extraction processes of this production are delignification, bleaching and hydrolysis. In order to produce NCC, raw rice husk (RRH) was grinded and converted to powder form. Powder rice husk was obtained by sieving and the particles in the 75-710 μm size range was used for experimental work. The production of NCC was conducted into the jacketed glass reactor at 80 ˚C temperature under predetermined experimental conditions. In this work NaOH (4M) solution was used for delignification process. After certain experimental time delignified powder RH was collected from the reactor then washed, bleached and finally hydrolyzed in order to degrade cellulose to nanocrystalline cellulose (NCC). For bleaching and hydrolysis processes NaOCl (20%) and H2SO4 (4M) solutions were used, respectively. The resultant products from hydrolysis was neutralized by buffer solution and analyzed by FTIR, XRD, SEM, AFM and TEM. From the analysis, NCC has been identified successfully and the particle dimension has been confirmed to be in the range of 20-50 nm. From XRD results, the crystallinity of NCC was found to be approximately 45%.

Keywords: nanocrystalline cellulose, NCC, rice husk, biomass, chemical extraction

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Authors: Kessara Seneesrisakul, Erdogan Gulari, Sumaeth Chavadej

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The complex structure of lignocellulose leads to great difficulties in converting it to fermentable sugars for the ethanol production. The major hydrolysis impediments are the crystallinity of cellulose and the lignin content. To improve the efficiency of enzymatic hydrolysis, microbial pretreatment of corncob was investigated using two bacterial strains of Bacillus subtilis A 002 and Cellulomonas sp. TISTR 784 (expected to break open the crystalline part of cellulose) and lignin-degrading fungus, Phanerochaete sordida SK7 (expected to remove lignin from lignocellulose). The microbial pretreatment was carried out with each strain under its optimum conditions. The pretreated corncob samples were further hydrolyzed to produce reducing glucose with low amounts of commercial cellulase (25 U•g-1 corncob) from Aspergillus niger. The corncob samples were determined for composition change by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscope (SEM). According to the results, the microbial pretreatment with fungus, P. sordida SK7 was the most effective for enhancing enzymatic hydrolysis, approximately, 40% improvement.

Keywords: corncob, enzymatic hydrolysis, glucose, microbial pretreatment

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Authors: Sakinul Islam, Nhol Kao, Sati Bhattacharya, Rahul Gupta

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Nanocrystalline cellulose (NCC) has been widely used for miscellaneous applications due to its superior properties over other nanomaterials. However, the major problems associated with the production of NCC are long reaction time, low production rate and inefficient process. The mass production of NCC within a short period of time is still a great challenge. The main objective of this study is to produce NCC from rice husk agro waste biomass from a high pressure delignification process (HPDP), followed by bleaching and hydrolysis processes. The HPDP has not been explored for NCC production from rice husk biomass (RHB) until now. In order to produce NCC, powder rice husk (PRH) was placed into a stainless steel reactor at 80 ˚C under 5 bars. Aqueous solution of NaOH (4M) was used for the dissolution of lignin and other amorphous impurities from PRH. After certain experimental times (1h, 3.5h and 6h), bleaching and hydrolysis were carried out on delignified samples. NaOCl (20%) and H2SO4 (4M) solutions were used for bleaching and hydrolysis processes, respectively. The NCC suspension from hydrolysis was sonicated and neutralized by buffer solution for various characterisations. Finally NCC suspension was dried and analyzed by FTIR, XRD, SEM, AFM and TEM. The chemical composition of NCC and PRH was estimated by TAPPI (Technical Association of Pulp and Paper Industry) standard methods to observe the product purity. It was found that, the 6h of the HPDP was more efficient to produce good quality NCC than that at 1h and 3.5h due to low separation of non-cellulosic components from RHB. The analyses indicated the crystallinity of NCC to be 71 %, particle size of 20-50 nm (diameter) and 100-200 nm in length.

Keywords: nanocrystalline cellulose, NCC, high pressure delignification, bleaching, hydrolysis, agro-waste biomass

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Authors: Yogi Wibisono Budhi, Ferry Iskandar, Veinardi Suendo, Muhammad Fakhrudin, Neng Tresna Umi Culsum

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Oil palm empty fruit bunch (OPEFB), an abundant agro-waste in Indonesia, is being studied as raw material of Cellulose Nanocrystals (CNC) synthesis. Instead of conventional acid mineral, phosphotungstic acid (H₃PW₁₂O₄₀, HPW) was used to hydrolyze cellulose due to recycling ability and easy handling. Before hydrolysis process, dried EFB was treated by 4% NaOH solution at 90oC for 2 hours and then bleached using 2% NaClO₂ solution at 80oC for 3 hours to remove hemicellulose and lignin. Hydrolysis reaction parameters such as temperature, acid concentration, and reaction time were optimized with fixed solid-liquid ratio of 1:40. Response surface method was used for experimental design to determine the optimum condition of each parameter. HPW was extracted from the mixed solution and recycled with diethyl ether. CNC was separated from the solution by centrifuging and washing with distilled water and ethanol to remove degraded sugars and unreacted celluloses. In this study, pulp from dried EFB produced 44.8% yield of CNC. Dynamic Light Scattering (DLS) analysis showed that most of CNC equivalent diameter was 140 nm. Crystallinity index was observed at 73.3% using X-ray Diffraction (XRD) analysis. Thus, a green established process for the preparation of CNC was achieved.

Keywords: acid hydrolysis, cellulose nanocrystals, oil palm empty fruit bunch, phosphotungstic acid

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Authors: Lee Chiau Yeng, Norhayani Othman

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Regenerated cellulose fiber is fabricated in the NaOH/urea aqueous solution. In this work, cellulose is dissolved in 7 .wt% NaOH/12 .wt% urea in the temperature of -12 °C to prepare regenerated cellulose. Thermal and structure properties of cellulose and regenerated cellulose was compared and investigated by Field Emission Scanning Electron Microscopy (FeSEM), Fourier Transform Infrared Spectroscopy (FTIR), X-Ray Diffraction (XRD), Thermogravimetric analysis (TGA), and Differential Scanning Calorimetry. Results of FeSEM revealed that the regenerated cellulose fibers showed a more circular shape with irregular size due to fiber agglomeration. FTIR showed the difference in between the structure of cellulose and the regenerated cellulose fibers. In this case, regenerated cellulose fibers have a cellulose II crystalline structure with lower degree of crystallinity. Regenerated cellulose exhibited better thermal stability than the cellulose.

Keywords: regenerated cellulose, cellulose, NaOH, urea

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Authors: Bahiru Tsegaye, Chandrajit Balomajumder, Partha Roy

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The limited supply and related negative environmental consequence of fossil fuels are driving researcher for finding sustainable sources of energy. Lignocellulose biomass like sorghum straw is considered as among cheap, renewable and abundantly available sources of energy. However, lignocellulose biomass conversion to bioenergy like bioethanol is hindered due to the reluctant nature of lignin in the biomass. Therefore, removal of lignin is a vital step for lignocellulose conversion to renewable energy. The aim of this study is to optimize microwave pretreatment conditions using design expert software to remove lignin and to release maximum possible polysaccharides from sorghum straw for efficient hydrolysis and fermentation process. Sodium hydroxide concentration between 0.5-1.5%, v/v, pretreatment time from 5-25 minutes and pretreatment temperature from 120-2000C were considered to depolymerize sorghum straw. The effect of pretreatment was studied by analyzing the compositional changes before and after pretreatments following renewable energy laboratory procedure. Analysis of variance (ANOVA) was used to test the significance of the model used for optimization. About 32.8%-48.27% of hemicellulose solubilization, 53% -82.62% of cellulose release, and 49.25% to 78.29% lignin solubilization were observed during microwave pretreatment. Pretreatment for 10 minutes with alkali concentration of 1.5% and temperature of 1400C released maximum cellulose and lignin. At this optimal condition, maximum of 82.62% of cellulose release and 78.29% of lignin removal was achieved. Sorghum straw at optimal pretreatment condition was subjected to enzymatic hydrolysis and fermentation. The efficiency of hydrolysis was measured by analyzing reducing sugars by 3, 5 dinitrisylicylic acid method. Reducing sugars of about 619 mg/g of sorghum straw were obtained after enzymatic hydrolysis. This study showed a significant amount of lignin removal and cellulose release at optimal condition. This enhances the yield of reducing sugars as well as ethanol yield. The study demonstrates the potential of microwave pretreatments for enhancing bioethanol yield from sorghum straw.

Keywords: cellulose, hydrolysis, lignocellulose, optimization

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Authors: Samira Gharehkhani, Seyed Farid Seyed Shirazi, Abdolreza Gharehkhani, Hooman Yarmand, Ahmad Badarudin, Rushdan Ibrahim, Salim Newaz Kazi

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Nanocrystalline cellulose (NCC) were prepared by acid hydrolysis and ultrasound treatment of bleached Acacia mangium fibers. The obtained rod-shaped nanocrystals showed a uniform size. The results showed that NCC with high crystallinity can be obtained using 64 wt% sulfuric acid. The effect of synthesis condition was investigated. Different reaction times were examined to produce the NCC and the results revealed that an optimum reaction time has to be used for preparing the NCC. Morphological investigation was performed using the transmission electron microscopy (TEM). Fourier transform infrared (FTIR) spectroscopy and thermogravimetric analysis (TGA) were performed. X-ray diffraction (XRD) analysis revealed that the crystallinity increased with successive treatments. The NCC suspension was homogeneous and stable and no sedimentation was observed for a long time.

Keywords: acid hydrolysis, nanocrystalline cellulose, nano material, reaction time

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Authors: Jitlada Chumee, Drenpen Seeburin

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The cellulose was extracted from pomelo peel and an etherification reaction used for converting cellulose to carboxymethyl cellulose (CMC). The pomelo peel was refluxed with 0.5 M HCl and 1 M NaOH solution at 90°C for 1 h and 2 h, respectively. The cellulose was bleached with calcium hypochlorite and used as precursor. The precursor was soaked in mixed solution between isopropyl alcohol and 40%w/v NaOH for 12 h. After that, chloroacetic acid was added and reacted at 55°C for 6 h. The optimum condition was 5 g of cellulose: 0.25 mole of NaOH : 0.07 mole of ClCH2COOH with 78.00% of yield. Moreover, the product had 0.54 of degree of substitution (DS).

Keywords: pomelo peel, carboxymethyl cellulose, bioplastic, extraction

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Authors: Deokyeong Choe, Jae Eun Nam, Young Hoon Roh, Chul Soo Shin

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The development of hydrogels with a high mechanical strength is important for numerous applications of hydrogels. As a material for tough hydrogels, cellulose has attracted much interest. However, cellulose cannot be melted and is very difficult to be dissolved in most solvents. Therefore, its dissolution in tetrabutylammonium fluoride/dimethyl sulfoxide (TBAF/DMSO) solvents has attracted researchers for chemical processing of cellulose. For this reason, studies about rheological properties of cellulose/TBAF/DMSO solution will provide useful information. In this study, viscosities of cellulose solutions prepared using different amounts of cellulose and TBAF in DMSO were measured. As expected, the viscosity of cellulose solution decreased with respect to the increasing volume of DMSO. The most viscose cellulose solution was achieved at a 1:1 mass ratio of cellulose to TBAF regardless of their contents in DMSO. At a 1:1 mass ratio of cellulose to TBAF, the formation of cellulose nanoparticles (467 nm) resulted in a dramatic increase in the viscosity, which led to the fabrication of 3D cellulose hydrogels.

Keywords: cellulose, TBAF/DMSO, viscosity, hydrogel

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Authors: Ahlam Said Al Azkawi, Nallusamy Sivakumar, Saif Nasser Al Bahri

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Diverse approaches and pathways are under development with the determination to develop cellulosic biofuels and other bio-products eventually at commercial scale in “bio-refineries”; however, the key challenge is mainly the high level of complexity in processing the feedstock which is complicated and energy consuming. To overcome the complications in utilizing the naturally occurring lignocellulose biomass, using waste paper as a feedstock for bio-production may solve the problem. Besides being abundant and cheap, bioprocessing of waste paper has evolved in response to the public concern from rising landfill cost from shrinking landfill capacity. Cardboard (CB) is one of the major components of municipal solid waste and one of the most important items to recycle. Although 50-70% of cardboard constitute is known to be cellulose and hemicellulose, the presence of lignin around them cause hydrophobic cross-link which physically obstructs the hydrolysis by rendering it resistant to enzymatic cleavage. Therefore, pretreatment is required to disrupt this resistance and to enhance the exposure of the targeted carbohydrates to the hydrolytic enzymes. Several pretreatment approaches have been explored, and the best ones would be those can influence cellulose conversion rates and hydrolytic enzyme performance with minimal or less cost and downstream processes. One of the promising strategies in this field is the application of surfactants, especially non-ionic surfactants. In this study, triton-X 100 was used as surfactants to treat cardboard prior enzymatic hydrolysis and compare it with acid treatment using 0.1% H2SO4. The effect of the surfactant enhancement was evaluated through its effect on hydrolysis rate in respect to time in addition to evaluating the structural changes and modification by scanning electron microscope (SEM) and X-ray diffraction (XRD) and through compositional analysis. Further work was performed to produce ethanol from CB treated with triton-X 100 via separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF). The hydrolysis studies have demonstrated enhancement in saccharification by 35%. After 72 h of hydrolysis, a saccharification rate of 98% was achieved from CB enhanced with triton-X 100, while only 89 of saccharification achieved from acid pre-treated CB. At 120 h, the saccharification % exceeded 100 as reducing sugars continued to increase with time. This enhancement was not supported by any significant changes in the cardboard content as the cellulose, hemicellulose and lignin content remained same after treatment, but obvious structural changes were observed through SEM images. The cellulose fibers were clearly exposed with very less debris and deposits compared to cardboard without triton-X 100. The XRD pattern has also revealed the ability of the surfactant in removing calcium carbonate, a filler found in waste paper known to have negative effect on enzymatic hydrolysis. The cellulose crystallinity without surfactant was 73.18% and reduced to 66.68% rendering it more amorphous and susceptible to enzymatic attack. Triton-X 100 has proved to effectively enhance CB hydrolysis and eventually had positive effect on the ethanol yield via SSF. Treating cardboard with only triton-X 100 was a sufficient treatment to enhance the enzymatic hydrolysis and ethanol production.

Keywords: cardboard, enhancement, ethanol, hydrolysis, treatment, Triton-X 100

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Authors: Rosli Mohd Yunus, Zianor Azrina Zianon Abdin, Mohammad Dalour Hossen Beg, Ridzuan Ramli

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Recently, nanocrystalline cellulose (NCC) has gained considerable interest as a promising biomaterial due to their outstanding properties such as high surface area, high mechanical properties, hydrophilicity, biocompatibility and biodegradability. The NCC also has good stability in water which is compatible for mixing of water based polymer solution or emulsions with NCC. Oil palm empty fruit bunch (EFB) contained different amount of lignocellulosic materials such as lignin, hemicellulose and cellulose. Cellulose is the most significant materials that can be extracted from EFB as nanocrystalline cellulose (NCC). In this work the nanocrystalline cellulose were produced through acid hydrolysis together with ultrasound technique. The morphology of NCC was characterized by TEM, thermal behavior has been studied with DSC, TGA analysis. Structural properties were illustrated X-Ray diffraction as well as FTIR. The hydrogel was produced using polyvinyl alcohol (PVA) with different concentration of NCC. The hydrogel composite was characterized by swelling ratio, crosslinking density, mechanical properties and morphology.

Keywords: nanocellulose, oil palm, hydrogel, water treatment

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Authors: Sherif M. A. S. Keshk, Hisham S. M. Abd-Rabboh, Mohamed S. Hamdy, Ibrahim H. A. Badr

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Microwave radiation was applied to synthesize nanoparticles of nickel oxide supported on pretreated cellulose with metal acetate in the presence of NaOH. Optimization, in terms of irradiation time and metal concentration, was investigated. FT-IR spectrum of cellulose/NiO spectrum shows a band at 445 cm^-1 that is related to the Ni–O stretching vibration of NiO6 octahedral in the cubic NiO structure. cellulose/NiO showed similar XRD pattern of cellulose I and exhibited sharpened reflection peak at 2q = 29.8°, corresponding to (111) plane of NiO, with two weak broad peaks at 48.5°, and 49.2°, representing (222) planes of NiO. XPS spectrum of mercerized cellulose/NiO composite showed did not show any peaks corresponding to Na ion.

Keywords: cellulose, mercerized cellulose, cellulose/zinc and nickeloxides composite, FTIR, XRD, XPS, SEM, Raman spectrum

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Authors: T. R. Bandara, H. Jaelani, G. J. Griffin

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The conversion of lignocellulosic waste materials, such as sugar cane bagasse, to biofuels such as ethanol has attracted significant interest as a potential element for transforming transport fuel supplies to totally renewable sources. However, the refractory nature of the cellulosic structure of lignocellulosic materials has impeded progress on developing an economic process, whereby the cellulose component may be effectively broken down to glucose monosaccharides and then purified to allow downstream fermentation. Ionic liquid (IL) treatment of lignocellulosic biomass has been shown to disrupt the crystalline structure of cellulose thus potentially enabling the cellulose to be more readily hydrolysed to monosaccharides. Furthermore, conventional hydrolysis of lignocellulosic materials yields byproducts that are inhibitors for efficient fermentation of the monosaccharides. However, selective extraction of monosaccharides from an aqueous/IL phase into an organic phase utilizing a combination of boronic acids and quaternary amines has shown promise as a purification process. Hydrolysis of sugar cane bagasse immersed in an aqueous solution with IL (1-ethyl-3-methylimidazolium acetate) was conducted at different pH and temperature below 100 ºC. It was found that the use of a high concentration of hydrochloric acid to acidify the solution inhibited the hydrolysis of bagasse. At high pH (i.e. basic conditions), using sodium hydroxide, catalyst yields were reduced for total reducing sugars (TRS) due to the rapid degradation of the sugars formed. For purification trials, a supported liquid membrane (SLM) apparatus was constructed, whereby a synthetic solution containing xylose and glucose in an aqueous IL phase was transported across a membrane impregnated with phenyl boronic acid/Aliquat 336 to an aqueous phase. The transport rate of xylose was generally higher than that of glucose indicating that a SLM scheme may not only be useful for purifying sugars from undesirable toxic compounds, but also for fractionating sugars to improve fermentation efficiency.

Keywords: biomass, bagasse, hydrolysis, monosaccharide, supported liquid membrane, purification

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Authors: Efri Mardawati, Ronny Purwadi, Made Tri Ari Penia Kresnowati, Tjandra Setiadi

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EFB are mainly composed of cellulose (≈ 43%), hemicellulose (≈ 23%) and lignin (≈20%). The palm oil empty fruit bunches (EFB) is the lignosellulosic waste from crude palm oil industries mainly compose of (≈ 43%), hemicellulose (≈ 23%) and lignin (≈20%). Xylan, a polymer made of pentose sugar xylose and the most abundant component of hemicellulose in plant cell wall. Further xylose can be used as a raw material for production of a wide variety of chemicals such as xylitol, which is extensively used in food, pharmaceutical and thin coating applications. Currently, xylose is mostly produced from xylan via chemical hydrolysis processes. However, these processes are normally conducted at a high temperature and pressure, which is costly, and the required downstream processes are relatively complex. As an alternative method, enzymatic hydrolysis of xylan to xylose offers an environmentally friendly biotechnological process, which is performed at ambient temperature and pressure with high specificity and at low cost. This process is catalysed by xylanolytic enzymes that can be produced by some fungal species such as Aspergillus niger, Penicillium crysogenum, Tricoderma reseei, etc. Fungal that will be used to produce crude xylanase enzyme in this study is T. Viride ITB CC L.67. It is the purposes of this research to study the influence of pretreatment of EFB for the enzymatic hydrolysis process, optimation of temperature and pH of the hydrolysis process, the influence of substrate and enzyme concentration to the enzymatic hydrolysis process, the dynamics of hydrolysis process and followingly to study the kinetics of this process. Xylose as the product of enzymatic hydrolysis process analyzed by HPLC. The results show that the thermal pretreatment of EFB enhance the enzymatic hydrolysis process. The enzymatic hydrolysis can be well approached by the Michaelis Menten kinetic model, and kinetic parameters are obtained from experimental data.

Keywords: oil palm empty fruit bunches (EFB), xylose, enzymatic hydrolysis, kinetic modelling

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Authors: Amtul Jamil Sami

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Apis mellifera is an insect of immense economic importance lives on rich carbohydrate diet including cellulose, nectar, honey and pollen. The carbohydrate metabolism in A mellifera has not been understood fully, as there are no data available, on the functional expression of cellulase gene. The cellulose hydrolyzing enzyme is required for the digestion of pollen cellulose wall, to release the important nutrients (amino acids, minerals, vitamins etc.) from the pollen. A dissection of Apis genome had revealed that there is one gene present for the expression of endo-beta-1,4-glucanase, for cellulose hydrolysis. In the presented work, functional expression of endo-beta-1,4 glucanase gene is reported. Total soluble proteins of the honey bee were isolated and were tested cellulose hydrolyzing enzyme activity, using carboxy-methyl cellulose, as a substrate. A mellifera proteins were able to hydrolyze carboxy-methyl cellulose, confirming its endo- type mode of action. Endo beta-1,4 glucanase enzyme was only present in the gut tissues, no activity was detected in the salivary glands. The pH optima of the enzyme were in the acidic pH range of 4-5-5-0, indicating its metabolic role in the acidic stomach of A mellifera. The reported enzyme is unique, as endo-beta- 1,4 glucanase was able to generate non reducing sugar, as an end product. The results presented, are supportive to the information that the honey bee is capable of producing its novel endo-beta-1,4 glucanase. Further it could be helpful, in understanding, the carbohydrate metabolism in A mellifera.

Keywords: honey bees, Endo-beta 1, 4- glucanase, Apis mellifera, functional expression

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Authors: Amanda Gregorim Fernandes, Lorena Cardoso Cintra, Rosalia Santos Amorim Jesuino, Fabricia Paula De Faria, Marcio José Poças Fonseca

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Bioethanol is one of the most promising biofuels and able to replace fossil fuels and reduce its different environmental impacts and can be generated from various agroindustrial waste. The Brazil is in first place in bioethanol production to be the largest producer of sugarcane. The bagasse sugarcane (SCB) has lignocellulose which is composed of three major components: cellulose, hemicellulose and lignin. Cellulose is a homopolymer of glucose units connected by glycosidic linkages. Among all species of Penicillium, Penicillium echinulatum has been the focus of attention because they produce high quantities of cellulase and the mutant strain 9A02S1 produces higher enzyme levels compared to the wild. Among the cellulases, the cellobiohydrolases enzymes are the main components of the cellulolytic system of fungi, and are also responsible for most of the potential hydrolytic in enzyme cocktails for the industrial processing of plant biomass and several cellobiohydrolases Penicillium had higher specific activity against cellulose compared to CBH I from Trichoderma reesei. This fact makes it an interesting pattern for higher yields in the enzymatic hydrolysis, and also they are important enzymes in the hydrolysis of crystalline regions of cellulose. Therefore, finding new and more active enzymes become necessary. Meanwhile, β-glycosidases act on soluble substrates and are highly dependent on cellobiohydrolases and endoglucanases action to provide the substrate in the hydrolysis of the biomass, but the cellobiohydrolases and endoglucanases are highly dependent β-glucosidases to maintain efficient hydrolysis. Thus, there is a need to understand the structure-function relationships that govern the catalytic activity of cellulolytic enzymes to elucidate its mechanism of action and optimize its potential as industrial biocatalysts. To evaluate the enzyme β-glucosidase of Penicillium echinulatum (PeBGL1) the gene was synthesized from the assembly sequence from a library in induction conditions and then the PeBGL1 gene was cloned in the vector pPICZαA and transformed into P. pastoris GS115. After processing, the producers of PeBGL1 were analyzed for enzyme activity and protein profile where a band of approximately 100 kDa was viewed. It was also carried out the zymogram. In partial characterization it was determined optimum temperature of 50°C and optimum pH of 6,5. In addition, to increase the secreted recombinant PeBGL1 production by Pichia pastoris, three parameters of P. pastoris culture medium were analysed: methanol, nitrogen source concentrations and the inoculum size. A 23 factorial design was effective in achieving the optimum condition. Altogether, these results point to the potential application of this P. echinulatum β-glucosidase in hydrolysis of cellulose for the production of bioethanol.

Keywords: bioethanol, biotechnology, beta-glucosidase, penicillium echinulatum

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Authors: Rajeev Ravindran, Amit K. Jaiswal

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Lignocellulose is the largest reservoir of inexpensive, renewable source of carbon. It is composed of lignin, cellulose and hemicellulose. Cellulose and hemicellulose is composed of reducing sugars glucose, xylose and several other monosaccharides which can be metabolised by microorganisms to produce several value added products such as biofuels, enzymes, aminoacids etc. Enzymatic treatment of lignocellulose leads to the release of monosaccharides such as glucose and xylose. However, factors such as the presence of lignin, crystalline cellulose, acetyl groups, pectin etc. contributes to recalcitrance restricting the effective enzymatic hydrolysis of cellulose and hemicellulose. In order to overcome these problems, pre-treatment of lignocellulose is generally carried out which essentially facilitate better degradation of lignocellulose. A range of pre-treatment strategy is commonly employed based on its mode of action viz. physical, chemical, biological and physico-chemical. However, existing pretreatment strategies result in lower sugar yield and formation of inhibitory compounds. In order to overcome these problems, we proposes a novel pre-treatment, which utilises the superior oxidising capacity of alkaline potassium permanganate assisted by ultra-sonication to break the covalent bonds in spent coffee waste to remove recalcitrant compounds such as lignin. The pre-treatment was conducted for 30 minutes using 2% (w/v) potassium permanganate at room temperature with solid to liquid ratio of 1:10. The pre-treated spent coffee waste (SCW) was subjected to enzymatic hydrolysis using enzymes cellulase and hemicellulase. Shake flask experiments were conducted with a working volume of 50mL buffer containing 1% substrate. The results showed that the novel pre-treatment strategy yielded 7 g/L of reducing sugar as compared to 3.71 g/L obtained from biomass that had undergone dilute acid hydrolysis after 24 hours. From the results obtained it is fairly certain that ultrasonication assists the oxidation of recalcitrant components in lignocellulose by potassium permanganate. Enzyme hydrolysis studies suggest that ultrasound assisted alkaline potassium permanganate pre-treatment is far superior over treatment by dilute acid. Furthermore, SEM, XRD and FTIR were carried out to analyse the effect of the new pre-treatment strategy on structure and crystallinity of pre-treated spent coffee wastes. This novel one-step pre-treatment strategy was implemented under mild conditions and exhibited high efficiency in the enzymatic hydrolysis of spent coffee waste. Further study and scale up is in progress in order to realise future industrial applications.

Keywords: spent coffee waste, alkaline potassium permanganate, ultra-sonication, physical characterisation

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Authors: Ridzuan Ramli, Norhafzan Junadi, Mohammad D.H. Beg, Rosli M. Yunus

Abstract:

In this study, microcrystalline cellulose (MCC) was extracted from oil palm empty fruit bunch (EFB) cellulose which was earlier isolated from oil palm EFB fibre. In order to isolate the cellulose, the chlorination method was carried out. Then, the MCC was prepared by simultaneous ultrasonic and alkali treatment from the isolated α-cellulose. Based on mass balance calculation, the yields for MCC obtained from EFB was 44%. For fiber characterization, it is observed that the chemical composition of the hemicellulose and lignin for all samples decreased while composition for cellulose increased. The structural property of the MCC was studied by X-ray diffraction (XRD) method and the result shows that the MCC produced is a cellulose-I polymorph, with 73% crystallinity.

Keywords: oil palm empty fruit bunch, microcrystalline cellulose, ultrasonic, alkali treatment, x-ray diffraction

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Authors: Z. Kassab, A. Aboulkas, A. Barakat, M. El Achaby

Abstract:

The red algae are available enormously around the world and their exploitation for the production of agar product has become as an important industry in recent years. However, this industrial processing of red algae generated a large quantity of solid fibrous wastes, which constitute a source of a serious environmental problem. For this reason, the exploitation of this solid waste would help to i) produce new value-added materials and ii) to improve waste disposal from environment. In fact, this solid waste can be fully utilized for the production of cellulose microfibers and nanocrystals because it consists of large amount of cellulose component. For this purpose, the red algae waste was chemically treated via alkali, bleaching and acid hydrolysis treatments with controlled conditions, in order to obtain pure cellulose microfibers and cellulose nanocrystals. The raw product and the as-extracted cellulosic materials were successively characterized using serval analysis techniques, including elemental analysis, X-ray diffraction, thermogravimetric analysis, infrared spectroscopy and transmission electron microscopy. As an application, the as extracted cellulose nanocrystals were used as nanofillers for the production of polymer-based composite films with improved thermal and tensile properties. In these composite materials, the adhesion properties and the large number of functional groups that are presented in the CNC’s surface and the macromolecular chains of the polymer matrix are exploited to improve the interfacial interactions between the both phases, improving the final properties. Consequently, the high performances of these composite materials can be expected to have potential in packaging material applications.

Keywords: cellulose nanowhiskers, food packaging, polymer composites, red algae waste

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Authors: Asiya Rezzouq, Mehdi El Bouchti, Omar Cherkaoui, Sanaa Majid, Souad Zyade

Abstract:

In recent years, there has been a steady increase in the exploration of new renewable and non-conventional sources for the production of biodegradable nanomaterials. Nature harbours valuable cellulose-rich materials that have so far been under-exploited and can be used to create cellulose derivatives such as cellulose microfibres (CMFs) and cellulose nanocrystals (CNCs). These unconventional sources have considerable potential as alternatives to conventional sources such as wood and cotton. By using agricultural waste to produce these cellulose derivatives, we are responding to the global call for sustainable solutions to environmental and economic challenges. Responsible management of agricultural waste is increasingly crucial to reducing the environmental consequences of its disposal, including soil and water pollution, while making efficient use of these untapped resources. In this study, the main objective was to extract cellulose nanocrystals (CNC) from melon plant residues using methods that are both efficient and sustainable. To achieve this high-quality extraction, we followed a well-defined protocol involving several key steps: pre-treatment of the residues by grinding, filtration and chemical purification to obtain high-quality (CMF) with a yield of 52% relative to the initial mass of the melon plant residue. Acid hydrolysis was then carried out using phosphoric acid and sulphuric acid to convert (CMF) into cellulose nanocrystals. The extracted cellulose nanocrystals were subjected to in-depth characterization using advanced techniques such as transmission electron microscopy (TEM), thermogravimetric analysis (TGA), Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction. The resulting cellulose nanocrystals have exceptional properties, including a large specific surface area, high thermal stability and high mechanical strength, making them suitable for a variety of applications, including as reinforcements for composite materials. In summary, the study highlights the potential for recovering agricultural melon waste to produce high-quality cellulose nanocrystals with promising applications in industry, nanotechnology, and biotechnology, thereby contributing to environmental and economic sustainability.

Keywords: cellulose, melon plant residues, cellulose nanocrystals, properties, applications, composite materials

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Authors: Rungsinee Sothornvit, Pattrathip Rodsamran

Abstract:

Rice stubble consists of a high content of cellulose and can be synthesized as a cellulose derivative such as carboxymethyl cellulose (CMC) to value added products from agricultural waste. Therefore, the synthesis conditions and characterization the properties of CMC from rice stubble (CMCr) were investigated. Hemicellulose and lignin were first removed from the rice stubble using 10% NaOH at 55 C for 3 h and 5% NaOCl at 75 C for 15 min, respectively. Rice stubble cellulose was swollen in 30% NaOH and isopropanol as a solvent. The content of chloroacetic acid (5–7 g in 5 g of alkali cellulose), reaction temperature (50 and 70 C) and time (180, 270 and 360 min) were explored to obtain CMC. It was found that synthesis conditions did not affect significantly on moisture content and pH of CMCr. The best quality of CMCr was synthesized by using 7 g of chloroacetic acid and reacted at 50 C for 180 min based on 5 g of rice stubble cellulose. Degree of substitution (DS), viscosity and purity of CMCr were 0.64, 36.03 cP and 90.18 %, respectively. Furthermore, Fourier transform infrared (FT–IR) spectroscopy confirmed the presence of carboxymethyl substituents. CMCr was categorized in commercial scale as a low viscosity material and it can be used as film forming packaging materials for food and pharmaceutical product applications.

Keywords: rice stubble, cellulose, carboxymethyl cellulose, degree of substitution, purity

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Authors: Anna Trusek-Holownia, Andrzej Noworyta

Abstract:

Using an enzyme of known specificity the hydrolysis of protein was carried out in a controlled manner. The aim was to obtain oligopeptides being the so-called active peptides or their direct precursors. An original way of expression of the protein hydrolysis kinetics was introduced. Peptide bonds contained in the protein were recognized as a diverse-quality substrate for hydrolysis by the applied protease. This assumption was positively verified taking as an example the hydrolysis of albumin by thermolysin. Peptide linkages for this system should be divided into at least four groups. One of them is a group of bonds non-hydrolyzable by this enzyme. These that are broken are hydrolyzed at a rate that differs even by tens of thousands of times. Designated kinetic constants were k'F = 10991.4 L/g.h, k'M = 14.83L/g.h, k'S about 10-1 L/g.h for fast, medium and slow bonds, respectively. Moreover, a procedure for unfolding of the protein, conducive to the improved susceptibility to enzymatic hydrolysis (approximately three-fold increase in the rate) was proposed.

Keywords: peptide bond hydrolysis, kinetics, enzyme specificity, biologically active peptides

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Authors: Ghazi Faisal Najmuldeen, Noridah Abdullah, Mimi Sakinah

Abstract:

Bioethanol is an alcohol made by fermentation, mostly from carbohydrates, Cellulosic biomass, derived from non-food sources, such as castor shell waste, is also being developed as a feedstock for ethanol production Cellulose extracted from biomass sources is considered the future feedstock for many products due to the availability and eco-friendly nature of cellulose. In this study, castor shell (CS) biowaste resulted from the extraction of Castor oil from castor seeds was evaluated as a potential source of cellulose. The cellulose was extracted after pretreatment process was done on the CS. The pretreatment process began with the removal of other extractives from CS, then an alkaline treatment, bleaching process with hydrogen peroxide, and followed by a mixture of acetic and nitric acids. CS cellulose was analysed by infrared absorption spectroscopy (FTIR), scanning electron microscopy (SEM), X-ray diffraction (XRD), and thermogravimetric analysis (TGA). The result showed that the overall process was adequate to produce cellulose with high purity and crystallinity from CS waste. The cellulose was then hydrolyzed to produce glucose and then fermented to bioethanol.

Keywords: bioethanol, castor shell, cellulose, biowaste

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Authors: Jyosthna Khanna Goli, Shaik Naseeruddin, Hameeda Bee

Abstract:

Employing lignocellulosic biomass in fermentative production of xylitol and bioethanol is gaining interest as it is renewable, cheap, and abundantly available. Xylitol is a polyol, gaining its importance in the food and pharmacological industry due to its low calorific value and anti-cariogenic nature. Bioethanol from lignocellulosic biomass is widely accepted as an alternative fuel for transportation with reduced CO₂ emissions, thus reducing the greenhouse effect. Typha latifolia, an aquatic weed, was found to be promising lignocellulosic substrate as it posses a high amount of sugars and does not compete with arable lands and interfere with food and feed competition. In the present study, xylose from hemicellulosic fraction of typha is converted to xylitol by isolate Jfh5 (Candida. tropicalis) and cellulose part to ethanol using Saccharomyces cerevisiaeVS3. Initially, alkali pretreatment of typha using sodium hydroxide, potassium hydroxide, ammonium hydroxide, calcium hydroxide, sodium bisulphate and sodium dithionate for overnight (18h) at room temperature (28 ± 2°C), resulted in maximum delignification of 75% with 2% (v/v) sodium bisulphate. Later, pretreated biomass was subjected to acid hydrolysis with 1%, 1.5%, 2%, and 3% H₂SO₄ at 110 °C and 121°C for 30 and 60 min, respectively. 2% H₂SO₄ at 121°C for 60 min was found to release 13.5 g /l sugars, which on detoxification and fermentation produced 8.1g/l xylitol with yield and productivity of 0.65g/g and 0.112g/l/h respectively. Further enzymatic hydrolysis of the residual substrate obtained after acid hydrolysis released 11g/l sugar, which on fermentation with VS3 produced 4.9g/l ethanol with yield and productivity of 0.22g/g and 0.136g/l/h respectively.

Keywords: delignification, xylitol, bioethanol, acid hydrolysis, enzyme hydrolysis

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Authors: Fadilah Fadilah, Rochmadi Rochmadi, Siti Syamsiah, Djagal W. Marseno

Abstract:

Glucomannan can be found in the tuber of porang together with starch and proteinaceous components which were regarded as impurities. An enzymatic process for obtaining higher glucomannan content from Porang flour have been conducted. Papain was used for hydrolysing proteinaceous components in Porang flour which was conducted after a simultaneous extraction of glucomannan and enzymatic starch hydrolysis. Three variables affecting the rate were studied, i.e. temperature, the amount of enzyme and the stirring speed. The ninhydrin method was used to determine degree of protein hydrolysis. Results showed that the rising of degree of hydrolysis were fast in the first ten minutes of the reaction and then proceeded slowly afterward. The optimum temperature for hydrolysis was 60 ^oC. Increasing the amount of enzyme showed a remarkable effect to degree of hydrolysis, but the stirring speed had no significant effect. This indicated that the reaction controlled the rate of hydrolysis.

Keywords: degree of hydrolysis, ninhydrin, papain, porang flour, proteinaceous components

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Authors: Lukas Munster, Pavel Bazant, Ivo Kuritka

Abstract:

Zinc oxide sub-micro particles and metallic silver nano particles (Ag/ZnO) were deposited on micro crystalline cellulose surface by a fast, simple and environmentally friendly one-pot microwave assisted solvo thermal synthesis in an open vessel system equipped with an external reflux cooler. In order to increase the interaction between the surface of cellulose and the precipitated Ag/ZnO particles, oxidized form of cellulose (cellulose dialdehyde, DAC) prepared by periodate oxidation of micro crystalline cellulose was added to the reaction mixture of Ag/ZnO particle precursors and untreated micro crystalline cellulose. The structure and morphology of prepared hybrid powder materials were analysed by X-ray diffraction (XRD), energy dispersive analysis (EDX), scanning electron microscopy (SEM) and nitrogen absorption method (BET). Microscopic analysis of the prepared materials treated by ultra-sonication showed that Ag/ZnO particles deposited on the cellulose/DAC sample exhibit increased adhesion to the surface of the cellulose substrate which can be explained by the DAC adhesive effect in comparison with the material prepared without DAC addition.

Keywords: microcrystalline cellulose, microwave synthesis, silver nanoparticles, zinc oxide sub-microparticles, cellulose dialdehyde

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Authors: Surasak Siripornadulsil, Nutt Poomai, Wilailak Siripornadulsil

Abstract:

Due to a high ethanol demand, the approach for effective ethanol production is important and has been developed rapidly worldwide. Several agricultural wastes are highly abundant in celluloses and the effective cellulose enzymes do exist widely among microorganisms. Accordingly, the cellulose degradation using microbial cellulose to produce a low-cost substrate for ethanol production has attracted more attention. In this study, the cellulose producing bacterial strain has been isolated from rich straw and identified by 16S rDNA sequence analysis as Acinetobacter sp. KKU44. This strain is able to grow and exhibit the cellulose activity. The optimal temperature for its growth and cellulose production is 37 °C. The optimal temperature of bacterial cellulose activity is 60 °C. The cellulose enzyme from Acinetobacter sp. KKU44 is heat-tolerant enzyme. The bacterial culture of 36 h. showed highest cellulose activity at 120 U/mL when grown in LB medium containing 2% (w/v). The capability of Acinetobacter sp. KKU44 to grow in cellulosic agricultural wastes as a sole carbon source and exhibiting the high cellulose activity at high temperature suggested that this strain could be potentially developed further as a cellulose degrading strain for a production of low-cost substrate used in ethanol production.

Keywords: cellulose enzyme, bagasse, rice straw, rice husk, acinetobacter sp. KKU44

Procedia PDF Downloads 283