Search results for: degree of hydrolysis

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

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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: Anna Trusek-Holownia, Andrzej Noworyta

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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: 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: Mohamed A. Deyab, Omnia A. A. El-Shamy

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The purpose of this study is to reveal on the systematic evaluation of hydrogen production by aluminum hydrolysis in alkaline solutions containing different surfactants using hydrogen evolution measurements and supplemented by scan electron microscope (SEM) and energy dispersive X-ray analysis (EDX). It has been demonstrated that when alkaline concentration and solution temperature rise, the rate of H2 generation and, consequently, aluminum hydrolysis also rises. The addition of nonionic and cationic surfactants solution retards the rate of H2 production. The work is a promising option for carbon-free hydrogen production from renewable resources.

Keywords: energy, hydrogen, hydrolysis, surfactants

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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: 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: Jamal Abd Awadallak, Thiago Olinek Reinehr, Eduardo Raizer, Deise Molinari, Edson Antonio, Camila da Silva da Silva

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The hydrolysis of soy oil catalyzed by 1,3-specific enzyme (Lecitase Ultra) in a well-stirred bioreactor was studied. Two forms of applications of the ultrasound were evaluated aiming to increase reaction rates, wherein the use of probe ultrasound associated with the use of surfactant to pre-emulsify the substrate showed the best results. Two different reaction periods were found: the first where the ultrasound has great influence on reaction rates, and the second where ultrasound influence is minimal. Studies on the time of pre-emulsification, surfactant concentration and enzyme concentration showed that the initial rate of hydrolysis depends on the interfacial area between the oil phase and the aqueous phase containing the enzyme.

Keywords: specific enzyme, free fatty acids, Hydrolysis, lecitase ultra, ultrasound

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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: Sánchez Acuña, Juan Camilo, Granados Gómez, Mildred Magaly, Navarrete Rodríguez, Luisa Fernanda

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Nowadays, the main biofuels source production as bioethanol is food crops. This means a high competition between foods and energy production. For this reason, it is necessary to take into account the use of new raw materials friendly to the environment. The main objective of this paper is to evaluate the potential of the agro-industrial banana crop residues in the production of bioethanol. A factorial design of 2⁴ was used, the design has variables such as pH, time and concentration of hydrolysis, another variable is the time of fermentation that is of 7 or 15 days. In the hydrolysis phase, the pH is acidic (H₂SO₄) or basic (NaOH), the time is 30 or 15 minutes and the concentration is 0.1 or 0.5 M. It was observed that basic media, low concentrations, fermentation, and higher pretreatment times produced better performance in terms of biofuel obtained.

Keywords: bioethanol, biofuels, banana waste, hydrolysis

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Authors: Abinaya Balasubramanian, Satyabrata Ghosh, Satyahari Dey

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Non-Digestible Oligosaccharides(NDOs) are low molecular weight carbohydrates with degree of polymerization (DP) 3-20, that are delivered intact to the large intestine. NDOs are gaining attention as effective prebiotic molecules that facilitate prevention and treatment of several chronic diseases. Recently, NDOs are being obtained by cleaving complex polysaccharides as it results in high yield and also as the former tend to display greater bioactivity. Thelebolus sp. IITKGP BT-12, a recently identified psychrophilic, Ascomycetes fungus has been reported to produce a bioactive extracellular polysaccharide(EPS). The EPS has been proved to possess strong prebiotic activity and anti- proliferative effects. The current study is an attempt to identify and optimise the most suitable method for hydrolysis of the above mentioned novel EPS into NDOs, and further purify and characterise the same. Among physical, chemical and enzymatic methods, enzymatic hydrolysis was identified as the best method and the optimum hydrolysis conditions obtained using response surface methodology were: reaction time of 24h, β-(1,3) endo-glucanase concentration of 0.53U and substrate concentration of 10 mg/ml. The NDOs were purified using gel filtration chromatography and their molecular weights were determined using MALDI-TOF. The major fraction was found to have a DP of 7,8. The monomeric units of the NDOs were confirmed to be glucose using TLC and GCMS-MS analysis. The obtained oligosaccharides proved to be non-digestible when subjected to gastric acidity, salivary and pancreatic amylases and hence could serve as efficient prebiotics.

Keywords: characterisation, enzymatic hydrolysis, non-digestible oligosaccharides, response surface methodology

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Authors: Prasanna Belur, P. R. Ashwini, Sampath Charanyaa, I. Regupathi

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Indian oil sardine (Sardinella longiceps) are one of the richest and cheapest sources of n-3 polyunsaturated fatty acids (n-3 PUFA) such as Eicosapentaenoic acid (EPA) and Docosahexaenoic acid (DHA). The health benefits conferred by n-3 PUFA upon consumption, in the prevention and treatment of coronary, neuromuscular, immunological disorders and allergic conditions are well documented. Natural refined Indian Sardine oil generally contain about 25% (w/w) n-3 PUFA along with various unsaturated and saturated fatty acids in the form of mono, di, and triglycerides. Having high concentration of n-3 PUFA content in the glyceride form is most desirable for human consumption to avail maximum health benefits. Thus, enhancing the n-3 PUFA content while retaining it in the glyceride form with green technology is the need of the hour. In this study, refined Indian Sardine oil was subjected to selective hydrolysis by Candida antartica lipase to enhance n-3 PUFA content. The degree of hydrolysis and enhancement of n-3 PUFA content was estimated by determining acid value, Iodine value, EPA and DHA content (by Gas Chromatographic methods after derivitization) before and after hydrolysis. Various reaction parameters such as pH, temperature, enzyme load, lipid to aqueous phase volume ratio and incubation time were optimized by conducting trials with one parameter at a time approach. Incubating enzyme solution with refined sardine oil with a volume ratio of 1:1, at pH 7.0, for 60 minutes at 50 °C, with an enzyme load of 60 mg/ml was found to be optimum. After enzymatic treatment, the oil was subjected to refining to remove free fatty acids and moisture content using previously optimized refining technology. Enzymatic treatment at the optimal conditions resulted in 12.11 % enhancement in Degree of hydrolysis. Iodine number had increased by 9.7 % and n-3 PUFA content was enhanced by 112 % (w/w). Selective enhancement of n-3 PUFA glycerides, eliminating saturated and unsaturated fatty acids from the oil using enzyme is an interesting preposition as this technique is environment-friendly, cost effective and provide natural source of n-3 PUFA rich oil.

Keywords: Candida antartica, lipase, n-3 polyunsaturated fatty acids, sardine oil

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Authors: Sina Aghili, Ali Arasteh Nodeh

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Tamarisk usage as a new group of lignocelluloses material to produce fermentable sugars in bio-ethanol process was studied. The overall aim of this work was to establish the optimum condition for acid hydrolysis of this new material and a mathematical model predicting glucose release as a function of operation variable. Sulfuric acid concentration in the range of 20 to 60%(w/w), process temperature between 60 to 95oC, hydrolysis time from 120 to 240 min and solid content 5,10,15%(w/w) were used as hydrolysis conditions. HPLC was used to analysis of the product. This analysis indicated that glucose was the main fermentable sugar and was increased with time, temperature and solid content and acid concentration was a parabola influence in glucose production.The process was modeled by a quadratic equation. Curve study and model were found that 42% acid concentration, 15 % solid content and 90oC were in optimum condition.

Keywords: fermentable sugar, saccharification, wood, hydrolysis

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Authors: Salisu ibrahim, Abdalla Rababah

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In this research, we use Weighted G2-Multi-degree reduction of Bezier curve of degree n to a Bezier curve of degree m, m < n. The degree reduction of Bezier curves is used to represent a given Bezier curve of n by a Bezier curve of degree m, m < n. Exact degree reduction is not possible, and degree reduction is approximate process in nature. We derive a weighted degree reducing method that is geometrically continuous at the end points. Different norms will be considered, several error minimizations will be given. The proposed methods produce error function that are less than the errors of existing methods.

Keywords: Bezier curves, multiple degree reduction, geometric continuity, error function

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Authors: Z. Okay, M. Kalkan Erdoğan, M. Şahin, M. Saçak

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Polyacrylonitrile (PAN) is one of the most common and cheap fiber-forming polymers because of its high strength and high abrasion resistance properties. The result of alkaline hydrolysis of PAN fiber could be formed the products with conjugated sequences of –C=N–, acrylamide, sodium acrylate, and amidine. In this study, PAN fiber was hydrolyzed in a solution of sodium hydroxide, and this hydrolyzed PAN (HPAN) fiber was used to prepare conductive composite fiber by silver particles. The electrically conductive PAN fiber has the usage potential to produce variety of materials such as antistatic materials, life jackets and static charge reducing products. We monitored the change in the weight loss values of the PAN fiber with hydrolysis time. It was observed that a 60 % of weight loss was obtained in the fiber weight after 7h hydrolysis under the investigated conditions, but the fiber lost its fibrous structure. The hydrolysis time of 5h was found to be suitable in terms of preserving its fibrous structure. The change in the conductivity values of the composite with the preparation conditions such as hydrolysis time, silver ion concentration was studied. PAN fibers with different degrees of hydrolysis were treated with aqueous solutions containing different concentrations of silver ions by continuous stirring at 20 oC for 30 min, and the composite having the maximum conductivity of 2 S/cm could be prepared. The antibacterial property of the conductive HPAN fibers participated silver was also investigated. While the hydrolysis of the PAN fiber was characterized with FTIR and SEM techniques, the silver reduction process of the HPAN fiber was investigated with SEM and TGA-DTA techniques. The SEM micrographs showed that the surface of HPAN fiber was rougher and much more corroded than that of the PAN fiber. Composite, Conducting polymer, Fiber, Polyacrylonitrile.

Keywords: composite, conducting polymer, fiber, polyacrylonitrile

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Authors: Natalia Caldeira De Carvalho, Tassia Batista Pessato, Luis Gustavo R. Fernandes, Ricardo L. Zollner, Flavia Maria Netto

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Protein hydrolysates are ingredients of enteral diets and hypoallergenic formulas. Enzymatic hydrolysis is the most commonly used method for reducing the antigenicity of milk protein. The antigenicity and physicochemical characteristics of the protein hydrolysates depend on the reaction parameters. Among them, pH has been pointed out as of the major importance. Hydrolysis reaction in laboratory scale is commonly carried out under controlled pH (pH-stat). However, from the industrial point of view, controlling pH during hydrolysis reaction may be infeasible. This study evaluated the impact of pH control on the physicochemical properties and antigenicity of the hydrolysates of whey proteins with Alcalase. Whey protein isolate (WPI) solutions containing 3 and 7 % protein (w/v) were hydrolyzed with Alcalase 50 and 100 U g-1 protein at 60°C for 180 min. The reactions were carried out under controlled and uncontrolled pH conditions. Hydrolyses performed under controlled pH (pH-stat) were initially adjusted and maintained at pH 8.5. Hydrolyses carried out without pH control were initially adjusted to pH 8.5. Degree of hydrolysis (DH) was determined by OPA method, peptides profile was evaluated by HPLC-RP, and molecular mass distribution by SDS-PAGE/Tricine. The residual α-lactalbumin (α-La) and β-lactoglobulin (β-Lg) concentrations were determined using commercial ELISA kits. The specific IgE and IgG binding capacity of hydrolysates was evaluated by ELISA technique, using polyclonal antibodies obtained by immunization of female BALB/c mice with α-La, β-Lg and BSA. In hydrolysis under uncontrolled pH, the pH dropped from 8.5 to 7.0 during the first 15 min, remaining constant throughout the process. No significant difference was observed between the DH of the hydrolysates obtained under controlled and uncontrolled pH conditions. Although all hydrolysates showed hydrophilic character and low molecular mass peptides, hydrolysates obtained with and without pH control exhibited different chromatographic profiles. Hydrolysis under uncontrolled pH released, predominantly, peptides between 3.5 and 6.5 kDa, while hydrolysis under controlled pH released peptides smaller than 3.5 kDa. Hydrolysis with Alcalase under all conditions studied decreased by 99.9% the α-La and β-Lg concentrations in the hydrolysates detected by commercial kits. In general, β-Lg concentrations detected in the hydrolysates obtained under uncontrolled pH were significantly higher (p<0.05) than those detected in hydrolysates produced with pH control. The anti-α-La and anti-β-Lg IgE and IgG responses to all hydrolysates decreased significantly compared to WPI. Levels of specific IgE and IgG to the hydrolysates were below 25 and 12 ng ml-1, respectively. Despite the differences in peptide composition and α-La and β-Lg concentrations, no significant difference was found between IgE and IgG binding capacity of hydrolysates obtained with or without pH control. These results highlight the impact of pH on the hydrolysates characteristics and their concentrations of antigenic protein. Divergence between the antigen detection by commercial ELISA kits and specific IgE and IgG binding response was found in this study. This result shows that lower protein detection does not imply in lower protein antigenicity. Thus, the use of commercial kits for allergen contamination analysis should be cautious.

Keywords: allergy, enzymatic hydrolysis, milk protein, pH conditions, physicochemical characteristics

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Authors: M. Merabet-Khelassi, Z. Houiene, L. Aribi-Zouioueche, O. Riant

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Lipases (EC.3.1.1.3) are efficient biotools widely used for their remarkable chemo-, regio- and enantio-selectivity, especially, in kinetic resolution of racemates. They offer access to a large panel of enantiopure building blocks, such as secondary benzylic alcohols, commonly used as synthetic intermediates in pharmaceutical and agrochemical industries. Due to the stability of lipases in both water and organic solvents poor in water, they are able to catalyze both transesterifications of arylalkylcarbinols and hydrolysis of their corresponding acetates. The use of enzymatic hydrolysis in aqueous media still limited. In this presentation, we expose a practical methodology for the preparation of optically enriched acetates using a Candida antarctica lipase B-catalyzed hydrolysis in non-aqueous media in the presence of alkaline carbonate salts. The influence of several parameters which can intervene on the enzymatic efficiency such as the impact of the introduction of the carbonates salts, its amount and the nature of the alkaline earth metal are discussed. The obtained results show that the use of sodium carbonate with CAL-B enhances drastically both reactivity and selectivity of this immobilized lipase. In all cases, the resulting alcohols and remaining acetates are obtained in high ee values (up to > 99 %), and the selectivities reach (E > 500).

Keywords: alkaline-hydrolysis, enzymatic kinetic resolution, lipases, arylalkylcarbinol, non-aqueous media

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Authors: Achiraya Jiraprasertwong, Erdogan Gulari, Sumaeth Chavadej

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Among agricultural residues, sugarcane bagasse is one of the most convincing raw materials for the production of bioethanol due to its availability, and low cost through enzymatic hydrolysis and yeast fermentation. A pretreatment step is needed to enhance the enzymatic step. In this study, sugarcane bagasse (SCB), one of the most abundant agricultural residues in Thailand, was pretreated biologically with various microorganisms of white-rot fungus—Phanerochaete sordid (SK 7), Cellulomonas sp. (TISTR 784), and strain A 002 (Bacillus subtilis isolated from Thai higher termites). All samples with various microbial pretreatments were further hydrolyzed enzymatically by a commercial enzyme obtained from Aspergillus niger. The results showed that the pretreatment with the white-rot fungus gave the highest glucose concentration around two-fold higher when compared with the others.

Keywords: sugarcane bagasse, microorganisms, pretreatment, enzymatic hydrolysis

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Authors: Jyosthna Khanna Goli, Shaik Naseeruddin, Hameeda Bee

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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: Miguel Pereira, Lígia Pimentel, Manuela Pintado

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Animal blood is a major by-product of slaughterhouses and still represents a cost and environmental problem in some countries. To be eliminated, blood should be stabilised by cooking and afterwards the slaughterhouses must have to pay for its incineration. In order to reduce the elimination costs and valorise the high protein content the aim of this study was the optimization of hydrolysis conditions, in terms of enzyme ratio and time, in order to obtain hydrolysates with biological activity. Two enzymes were tested in this assay: pepsin and proteases from Cynara cardunculus (cardosins). The latter has the advantage to be largely used in the Portuguese Dairy Industry and has a low price. The screening assays were carried out in a range of time between 0 and 10 h and using a ratio of enzyme/reaction volume between 0 and 5%. The assays were performed at the optimal conditions of pH and temperature for each enzyme: 55 °C at pH 5.2 for cardosins and 37 °C at pH 2.0 for pepsin. After reaction, the hydrolysates were evaluated by FPLC (Fast Protein Liquid Chromatography) and tested for their antioxidant activity by ABTS method. FPLC chromatograms showed different profiles when comparing the enzymatic reactions with the control (no enzyme added). The chromatogram exhibited new peaks with lower MW that were not present in control samples, demonstrating the hydrolysis by both enzymes. Regarding to the antioxidant activity, the best results for both enzymes were obtained using a ratio enzyme/reactional volume of 5% during 5 h of hydrolysis. However, the extension of reaction did not affect significantly the antioxidant activity. This has an industrial relevant aspect in what concerns to the process cost. In conclusion, the enzymatic blood hydrolysis can be a better alternative to the current elimination process allowing to the industry the reuse of an ingredient with biological properties and economic value.

Keywords: antioxidant activity, blood, by-products, enzymatic hydrolysis

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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: Zhaojun Zheng, Yuanfa Liu, Jiaxin Li, Jinwei Li, Yong-jiang Xu, Chen Cao

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Black bean is an excellent protein source for preparing hydrolysates, which attract much attention due to their biological activity. The objective of this study was to characterize the physicochemical and antioxidant properties of black bean protein, hydrolyzed by ficin, bromelain or alcalase until 300 min of hydrolysis. Results showed that bromelain and alcalase hydrolysates possessed a higher degree of hydrolysis (DH) than that of ficin, thereby presenting different ultraviolet absorption, fluorescence intensity, and circular dichroism. Moreover, all hydrolysates possessed the capacity to scavenge DPPH radical with the lowest IC₅₀ of 21.11 µg/mL, as well as to chelate ferrous ion (Fe²⁺) with the IC₅₀ values ranging from 6.82 to 30.68 µg/mL. Intriguingly, the oxidation of linoleic acid, sunflower oil, and sunflower oil-in-water emulsion was remarkedly retarded by the three selected protein hydrolysates, especially by bromelain-treated protein hydrolysate, which might attribute to their high hydrophobicity and emulsifying properties. These findings can provide strong support for black bean protein hydrolysates to be employed in food products acting as natural antioxidant alternatives.

Keywords: antioxidant activity, black bean protein hydrolysate, emulsion physicochemical properties, sunflower oil

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Authors: Leo Paul Vaurs, Sonia Heaven, Charles Banks

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Municipal solid waste (MSW) is a virtually unlimited source of lignocellulosic material in the form of a waste paper/cardboard mixture which can be converted into fermentable sugars via cellulolytic enzyme hydrolysis in a biorefinery. The extraction of the lignocellulosic fraction and its preparation, however, are energy and water demanding processes. The waste water generated is a rich organic liquor with a high Chemical Oxygen Demand that can be partially cleaned while generating biogas in an Upflow Anaerobic Sludge Blanket bioreactor and be further re-used in the process. In this work, an experiment was designed to determine the critical contaminant concentrations in water affecting either anaerobic digestion or enzymatic hydrolysis by simulating multiple water re-circulations. It was found that re-using more than 16.5 times the same water could decrease the hydrolysis yield by up to 65 % and led to a complete granules desegregation. Due to the complexity of the water stream, the contaminant(s) responsible for the performance decrease could not be identified but it was suspected to be caused by sodium, potassium, lipid accumulation for the anaerobic digestion (AD) process and heavy metal build-up for enzymatic hydrolysis. The experimental data were incorporated into a Water Pinch technology based model that was used to optimize the water re-utilization in the modelled system to reduce fresh water requirement and wastewater generation while ensuring all processes performed at optimal level. Multiple scenarios were modelled in which sub-process requirements were evaluated in term of importance, operational costs and impact on the CAPEX. The best compromise between water usage, AD and enzymatic hydrolysis yield was determined for each assumed contaminant degradations by anaerobic granules. Results from the model will be used to build the first MSW based biorefinery in the USA.

Keywords: anaerobic digestion, enzymatic hydrolysis, municipal solid waste, water optimization

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Authors: S. R. Nayaka, Putta Swamy

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Graph polynomial is one of the algebraic representations of the Graph. The degree polynomial is one of the simple algebraic representations of graphs. The degree polynomial of a graph G of order n is the polynomial Deg(G, x) with the coefficients deg(G,i) where deg(G,i) denotes the number of vertices of degree i in G. In this article, we investigate the behavior of the roots of some families of Graphs in the complex field. We investigate for the graphs having only integral roots. Further, we characterize the graphs having single roots or having real roots and behavior of the polynomial at the particular value is also obtained.

Keywords: degree polynomial, regular graph, minimum and maximum degree, graph operations

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Authors: Ying Wang, Haiyan Xie, Aoming Zhang

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The degree distance of a graph is a graph invariant that is more sensitive than the Wiener index. In this paper, we calculate the vertex degree distances of one vertex union of the cycle and the star, and the degree distance of one vertex union of the cycle and the star. These results lay a foundation for further study on the extreme value of the vertex degree distances, and the distribution of the vertices with the extreme value in one vertex union of the cycle and the star.

Keywords: degree distance, vertex-degree-distance, one vertex union of a cycle and a star, graph

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Authors: I. Georgousopoulou, S. Vouyiouka, C. Papaspyrides

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The hydrolytic degradation of poly(butylene succinate) (PBS) was investigated when exposed to different humidity-temperature environments. To this direction different PBS grades were submitted to hydrolysis runs. Results indicated that the increment of hydrolysis temperature and relative humidity induced significant decrease in the molecular weight and thermal properties of the bioplastic. Τhe derived data can be considered to construct degradation kinetics based on carboxyl content variation versus time.

Keywords: hydrolytic degradation, physical ageing, poly(butylene succinate), polyester

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Authors: Burcu Kiren, Alattin CAkan, Nezihe Ayas

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Hydrogen will be arguably the best fuel in the future as it is the most abundant element in the universe. Hydrogen, as a fuel, is notably environmentally benign, sustainable and has high energy content compared to other sources of energy. It can be generated from both conventional and renewable sources. The hydrolysis reaction of metal hydrides provides an option for hydrogen production in the presence of a catalyst. In this study, Ni/dolomite catalyst was synthesized by the wet impregnation method for hydrogen production by hydrolysis reaction of sodium borohydride (NaBH4). Besides, the synthesized catalysts characterizations were examined by means of thermogravimetric analysis (TGA), Fourier-transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), Brunauer –Emmett – Teller (BET) and scanning electron microscopy (SEM). The influence of reaction temperature (25-75 °C), reaction time (15-60 min.), amount of catalyst (50-250 mg) and active metal loading ratio (20,30,40 wt.%) were investigated. The catalyst prepared with 30 wt.% Ni was noted as the most suitable catalyst, achieving of 35.18% H₂ and hydrogen production rate of 19.23 mL/gcat.min at 25 °C at reaction conditions of 5 mL of 0.25 M NaOH and 100 mg NaBH₄, 100 mg Ni/dolomite.

Keywords: sodium borohydride, hydrolysis, catalyst, Ni/dolomite, hydrogen

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Authors: Waranya Pongpaiboon, Warangkana Srichamnong, Supat Chaiyakul

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Enzymatic modification of rice flour can produce highly functional derivatives use in food industries. This study aimed to evaluate the physical properties and resistant starch content of rice flour residues hydrolyzed by α-amylase. Rice flour hydrolyzed by α-amylase (60 and 300 u/g) for 1, 24 and 48 hours were investigated. Increasing enzyme concentration and hydrolysis time resulted in decreased rice flour residue’s lightness (L*) but increased redness (a*) and yellowness (b*) of rice flour residues. The resistant starch content and peak viscosity increased when hydrolysis time increased. Pasting temperature, trough viscosity, breakdown, final viscosity, setback and peak time of the hydrolyzed flours were not significantly different (p>0.05). The morphology of native flour was smooth without observable pores and polygonal with sharp angles and edges. However, after hydrolysis, granules with a slightly rough and porous surface were observed and a rough and porous surface was increased with increasing hydrolyzed time. The X-ray diffraction patterns of native flour showed A-type configuration, which hydrolyzed flour showed almost 0% crystallinity indicated that both amorphous and crystalline structures of starch were simultaneously hydrolyzed by α-amylase.

Keywords: α-Amylase, enzymatic hydrolysis, pasting properties, resistant starch

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Authors: Dina M. Abd El-Aty, D. Aman, E. G. Zaki, Heba M. Salem

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A friendly environmental source of energy as hydrogen was produced by photo-hydrolysis of hydrogen storage material as sodium borohydride (NaBH4), which is non-toxic and stores a high percentage of hydrogen. The photoreaction was produced under visible light and nano-alumina as a catalyst. In this study, we use more economical and friendly environmental oil as a template to produce a nano-catalyst. The prepared catalyst was characterized by X-Ray diffraction, N2-adsorption-desorption, Fourier Transforms Infrared, Scanning Electron microscope and X-Ray Photoelectron Spectroscopy. Different parameters such as catalyst weight, NaBH4 weight and time of irradiation were studied to obtain a highly efficient photo-hydrolysis reaction. The reaction is pseudo-first order and the hydrogen production rate was determined as 1500 ml min-1 g-1 at the optimum conditions.

Keywords: photo-reaction, nano-alumina, hydrogen production, sodium borohydride, visible light

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Authors: Nurul Aqilah Mohd Zaini, Afroditi Chatzifragkou, Dimitris Charalampopoulos

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D-Lactic acid production is gaining increasing attention due to the thermostable properties of its polymer, Polylactic Acid (PLA). In this study, D-lactic acid was produced in microbial cultures using Lactobacillus coryniformis subsp. torquens as D-lactic acid producer and hydrolysates of Dried Distillers Grains with Solubles (DDGS) as fermentation substrate. Prior to fermentation, DDGS was first alkaline pretreated with 5% (w/v) NaOH, for 15 minutes (121oC/ ~16 psi). This led to the generation of DDGS solid residues, rich in carbohydrates and especially cellulose (~52%). The carbohydrate-rich solids were then subjected to enzymatic hydrolysis with Accellerase® 1500. For Separate Hydrolysis and Fermentation (SHF), enzymatic hydrolysis was carried out at 50oC for 24 hours, followed by fermentation of D-lactic acid at 37oC in controlled pH 6. The obtained hydrolysate contained 24 g/l glucose, 5.4 g/l xylose and 0.6 g/l arabinose. In the case of Simultaneous Saccharification and Fermentation (SSF), hydrolysis and fermentation were conducted in a single step process at 37oC in pH 5. The enzymatic hydrolysis of DGGS pretreated solids took place mostly during lag phase of L. coryniformis fermentation, with only a small amount of glucose consumed during the first 6 h. When exponential phase was started, glucose generation reduced as the microorganism started to consume glucose for D-lactic acid production. Higher concentrations of D-lactic acid were produced when SSF approach was applied, with 28 g/l D-lactic acid after 24 h of fermentation (84.5% yield). In contrast, 21.2 g/l D-lactic acid were produced when SHF was used. The optical pu rity of D-lactic acid produced from both experiments was 99.9%. Besides, approximately 2 g/l acetic acid was also generated due to lactic acid degradation after glucose depletion in SHF. SSF was proved an efficient towards DDGS ulilisation and D-lactic acid production, by reducing the overall processing time, yielding sufficient D-lactic acid concentrations without the generation of fermentation by-products.

Keywords: DDGS, alkaline pretreatment, SSF, D-lactic acid

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Authors: Anna Zimoch-Korzycka, Dominika Kulig, Andrzej Jarmoluk

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The aim of this study was to obtain chitooligosaccharides from chitosan with better functional properties using three different enzyme preparations and compare the products of enzymatic hydrolysis. Commercially available cellulase (CL), xylanase (X) and chitosanase (CS) preparations were used to investigate hydrolytic activity on chitosan (CH) with low molecular weight and DD of 75-85%. It has been reported that CL and X have side activities of other enzymes, such as β-glucanase or β-glucosidase. CS enzyme has a foreign activity of chitinase. Each preparation was used in 1000 U of activity and in the same reaction conditions. The degree of deacetylation and molecular weight of chitosan were specified using titration and viscometric methods, respectively. The hydrolytic activity of enzymes preparations on chitosan was monitored by dynamic viscosity measurement. After 4 h reaction with stirring, solutions were filtered and chitosan oligomers were isolated by methanol solution into two fractions: precipitate (A) and supernatant (B). A Fourier-transform infrared spectroscopy was used to characterize the structural changes of chitosan oligomers fractions and initial chitosan. Furthermore, the solubility of lyophilized hydrolytic mixture (C) and two chitooligomers fractions (A, B) of each enzyme hydrolysis was assayed. The antioxidant activity of chitosan oligomers was evaluated as DPPH free radical scavenging activity. The dynamic viscosity measured after addition of enzymes preparation to the chitosan solution decreased dramatically over time in the sample with X in comparison to solution without the enzyme. For mixtures with CL and CS, lower viscosities were also recorded but not as low as the ones with X. A and B fractions were characterized by the most similar viscosity obtained by the xylanase hydrolysis and were 15 mPas and 9 mPas, respectively. Structural changes of chitosan oligomers A, B, C and their differences related with various enzyme preparations used were confirmed. Water solubility of A fractions was not possible to filter and the result was not recorded. Solubility of supernatants was approximately 95% and was higher than hydrolytic mixture. It was observed that the DPPH radical scavenging effect of A, B, C samples is the highest for X products and was approximately 13, 17, 19% respectively. In summary, a mixture of chitooligomers may be useful for the design of edible protective coatings due to the improved biophysical properties.

Keywords: cellulase, xylanase, chitosanase, chitosan, chitooligosaccharides

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