Abstract: Pomegranate (Punica granatum L.) is an ancient fruit of great medical interest and rich source of antioxidants. Pesticides as dimethoate play a crucial role in the occurrence many diseases in plants, animal and human. Therefore the ability of Pomegranate (Punica granatum L.) to alleviate hepatotoxicity induced by organophosphate pesticide dimethoate was investigated. Albino male rats were divided randomly into 4 groups and kept at 7 animals per group in an environmentally controlled condition for 6 weeks. The first group was served as a control group (basal diet), the second group fed on basal diet supplemented with 5% freeze dried pomegranate seeds, the third group fed on 20 ppm dimethoate contaminated diet and the last group fed on dimethoate contaminated diet supplemented with 5% freeze dried pomegranate seeds. The results revealed that administration of dimethoate caused high significant increased in liver functions: alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) activities as well as lipid peroxide (malonaldhyde, MDA); on the other hand high significant decreased on glutathione (GSH), glutathione peroxidase (GPx), albumin and total protein were observed. However addition of 5% freeze dried pomegranate seeds significantly improved all previously mentioned parameters. These results indicate the dimethoate induced hepatotoxicity and highlight the protective effect of pomegranate seeds as a potential protective agent against dimethoate induced hepatotoxicity. This may be attributed to the powerful antioxidants (polyphenols, total phenols, and total flavonoids) which present in high levels in pomegranate as well as improving the immunity by activation of antioxidant enzymes GSH and GPx.
Abstract: Enzyme activity was evaluated in the intestine of
juvenile dourado (Salminus brasiliensis) fed with diets containing 0,
10 or 20% of lyophilized bovine colostrum (LBC) inclusion for either
30 or 60 days. The intestinal enzymes acid and alkaline phosphatase
(ACP and ALP, respectively), non-specific esterase (NSE), lipase
(LIP), dipeptidyl aminopeptidase IV (DAP IV) and leucine
aminopeptidase (LAP) were studied using histochemistry in four
intestinal segments (S1, S2, S3 and posterior intestine). Weak
proteolitic activity was observed in all intestinal segments for DAP
IV and LAP. The activity of NSE and LIP was also weak in all
intestines, except for the moderate activity of NSE in the S2 of 20%
LBC group after 30 days and in the S1 of 0% LBC group after 60
days. The ACP was detected only in the S2 and S3 of the 10% LBC
group after 30 days. Moderate and strong staining was observed in
the first three intestinal segments for ALP and weak activity in the
posterior intestine. The activity of DAP IV, LAP and ALP were also
present in the cytoplasm of the enterocytes. In the present results,
bovine colostrum feeding did not cause alterations in activity of
intestinal enzymes.
Abstract: Fibrin degradation is an important part in prevention
or treatment of intravascular thrombosis and cardiovascular diseases.
Plasmin like fibrinolytic enzymes has given new hope to patient with
cardiovascular diseases by treating fibrin aggregation related diseases
with traditional plasminogen activator which have many side effects.
Various researches involving wide range of sources for production of
fibrinolytic proteases, from bacteria, fungi, insects and fermented
foods. But few have looked into endophytic fungi as a potential
source. Sixteen (16) endophytic fungi were isolated from Hibiscus sp.
leaves from six different locations in Shah Alam, Selangor. Only two
endophytic fungi, FH3 and S13 showed positive fibrinolytic protease
activities. FH3 produced 5.78cm and S13 produced 4.48cm on Skim
Milk Agar after 4 days of incubation at 27°C. Fibrinolytic activity
was observed; 3.87cm and 1.82cm diameter clear zone on fibrin plate
of FH3 and S13 respectively. 18srRNA was done for identification of
the isolated fungi with positive fibrinolytic protease. S13 had the
highest similarity (100%) to that of Penicillium citrinum strain TG2
and FH3 had the highest similarity (99%) to that of Fusarium sp.
FW2PhC1, Fusarium sp. 13002, Fusarium sp. 08006, Fusarium
equiseti strain Salicorn 8 and Fungal sp. FCASAn-2. Media
composition variation showed the effects of carbon nitrogen on
protein concentration, where the decrement of 50% of media
composition caused drastic decrease in protease of FH3 from 1.081 to
0.056 and also S13 from 2.946 to 0.198.
Abstract: 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.
Abstract: Ferulic acid has widespread industrial potential by virtue of its antioxidant properties. However, it is partially soluble in aqueous media, limiting their usefulness in oil-based processes in food, cosmetic, pharmaceutical, and material industry. Therefore, modification of ferulic acid should be made by producing of more lipophilic derivatives. In this study, a preliminary investigation of lipase-catalyzed trans-esterification reaction of ethyl ferulate and olive oil was investigated. The reaction was catalyzed by immobilized lipase from Candida antarctica (Novozym 435), to produce ferulate ester, a sunscreen agent. A statistical approach of Response surface methodology (RSM) was used to evaluate the interactive effects of reaction temperature (40-80°C), reaction time (4-12 hours), and amount of enzyme (0.1-0.5 g). The optimum conditions derived via RSM were reaction temperature 60°C, reaction time 2.34 hours, and amount of enzyme 0.3 g. The actual experimental yield was 59.6% ferulate ester under optimum condition, which compared well to the maximum predicted value of 58.0%.
Abstract: The optimization of biological systems, which is a branch of metabolic engineering, has generated a lot of industrial and academic interest for a long time. In the last decade, metabolic engineering approaches based on mathematical optimizations have been used extensively for the analysis and manipulation of metabolic networks. In practical optimization of metabolic reaction networks, designers have to manage the nature of uncertainty resulting from qualitative characters of metabolic reactions, e.g., the possibility of enzyme effects. A deterministic approach does not give an adequate representation for metabolic reaction networks with uncertain characters. Fuzzy optimization formulations can be applied to cope with this problem. A fuzzy multi-objective optimization problem can be introduced for finding the optimal engineering interventions on metabolic network systems considering the resilience phenomenon and cell viability constraints. The accuracy of optimization results depends heavily on the development of essential kinetic models of metabolic networks. Kinetic models can quantitatively capture the experimentally observed regulation data of metabolic systems and are often used to find the optimal manipulation of external inputs. To address the issues of optimizing the regulatory structure of metabolic networks, it is necessary to consider qualitative effects, e.g., the resilience phenomena and cell viability constraints. Combining the qualitative and quantitative descriptions for metabolic networks makes it possible to design a viable strain and accurately predict the maximum possible flux rates of desired products. Considering the resilience phenomena in metabolic networks can improve the predictions of gene intervention and maximum synthesis rates in metabolic engineering. Two case studies will present in the conference to illustrate the phenomena.
Abstract: Protein kinases participate in a myriad of cellular
processes of major biomedical interest. The in vivo substrate
specificity of these enzymes is a process determined by several
factors, and despite several years of research on the topic, is still
far from being totally understood. In the present work, we have
quantified the contributions to the kinase substrate specificity of
i) the phosphorylation sites and their surrounding residues in the
sequence and of ii) the association of kinases to adaptor or scaffold
proteins. We have used position-specific scoring matrices (PSSMs),
to represent the stretches of sequences phosphorylated by 93 families
of kinases. We have found negative correlations between the number
of sequences from which a PSSM is generated and the statistical
significance and the performance of that PSSM. Using a subset
of 22 statistically significant PSSMs, we have identified specificity
determinant residues (SDRs) for 86% of the corresponding kinase
families. Our results suggest that different SDRs can function as
positive or negative elements of substrate recognition by the different
families of kinases. Additionally, we have found that human proteins
with known function as adaptors or scaffolds (kAS) tend to interact
with a significantly large fraction of the substrates of the kinases to
which they associate. Based on this characteristic we have identified
a set of 279 potential adaptors/scaffolds (pAS) for human kinases,
which is enriched in Pfam domains and functional terms tightly
related to the proposed function. Moreover, our results show that
for 74.6% of the kinase–pAS association found, the pAS colocalize
with the substrates of the kinases they are associated to. Finally, we
have found evidence suggesting that the association of kinases to
adaptors and scaffolds, may contribute significantly to diminish the
in vivo substrate crossed-specificity of protein kinases. In general, our
results indicate the relevance of several SDRs for both the positive
and negative selection of phosphorylation sites by kinase families and
also suggest that the association of kinases to pAS proteins may be
an important factor for the localization of the enzymes with their set
of substrates.
Abstract: Immobilization of lipase enzyme produced from palm oil mill effluent (POME) by the activated carbon (AC) among the low cost support materials was optimized. The results indicated that immobilization of 94% was achieved by AC as the most suitable support material. A sequential optimization strategy based on a statistical experimental design, including one-factor-at-a-time (OFAT) method was used to determine the equilibrium time. Three components influencing lipase immobilization were optimized by the response surface methodology (RSM) based on the face-centered central composite design (FCCCD). On the statistical analysis of the results, the optimum enzyme concentration loading, agitation rate and carbon active dosage were found to be 30 U/ml, 300 rpm and 8 g/L respectively, with a maximum immobilization activity of 3732.9 U/g-AC after 2 hrs of immobilization. Analysis of variance (ANOVA) showed a high regression coefficient (R2) of 0.999, which indicated a satisfactory fit of the model with the experimental data. The parameters were statistically significant at p
Abstract: The hydrolysis of lactose using β-galactosidase is one of the most promising biotechnological applications, which has wide range of potential applications in food processing industries. However, due to intracellular location of the yeast enzyme, and expensive extraction methods, the industrial applications of enzymatic hydrolysis processes are being hampered. The use of permeabilization technique can help to overcome the problems associated with enzyme extraction and purification of yeast cells and to develop the economically viable process for the utilization of whole cell biocatalysts in food industries. In the present investigation, standardization of permeabilization process of novel yeast isolate was carried out using a statistical model approach known as Response Surface Methodology (RSM) to achieve maximal b-galactosidase activity. The optimum operating conditions for permeabilization process for optimal β-galactosidase activity obtained by RSM were 1:1 ratio of toluene (25%, v/v) and ethanol (50%, v/v), 25.0 oC temperature and treatment time of 12 min, which displayed enzyme activity of 1.71 IU /mg DW.
Abstract: After cold pressing of pumpkin oil, the defatted oil cake (PUOC) was utilised as raw material for processing of bio-functional hydrolysates. In this study, the in vitro bioactivity of an alcalase (AH) and a pepsin hydrolysate (PH) prepared from the major pumpkin 12S globulin (cucurbitin) are compared. The hydrolysates were produced at optimum reaction conditions (temperature, pH) for the enzymes, during 60min. The bioactivity testing included antioxidant and angiotensin I converting enzyme inhibitory activity assays. The hydrolysates showed high potential as natural antioxidants and possibly antihypertensive agents in functional food or nutraceuticals. Additionally, preliminary studies have shown that both hydrolysates could exhibit modest α-amylase inhibitory activity, which indicates on their hypoglycemic potential.
Abstract: A process of conversion of flour from three varieties of cassava, namely Odongbo, ofege and TMS30752 to ethanol using α-amylase locally sourced from germinated unhusked paddy rice and yeast isolated from palm wine was developed. It involves the germination of paddy rice for a period of 15days to produce α-amylase for starch hydrolysis and isolation of yeast from palm wine for fermentation. The results showed that optimum amylase yield of “ofada” rice paddy was at 6th day germination which was 576.9ml/g. Ethanol yield for TMS30572 (440.3%) was significantly higher than “Odongbo” (160.2%) and “Ofege’’ (115.1%), Sugar conversion efficiency were 311.0%v/v, 268.2%v/v and 186.84%v/v for TMS30572, “Odongbo” and “Ofege” respectively. The ethanol boiling points were 78oC, 76oC and 80oC for TMS30572, “Odongbo” and “Ofege” respectively. This study showed that cassava varieties affects quality of ethanol produced and germination of “ofada” rice for 6 days ensures optimum production of crude amylase enzyme.
Abstract: The goal of presented work is the development phytoremediation method targeted to cleaning environment polluted with organochlorine pesticides, based on joint application of plants and microorganisms. For this aim the selection of plants and microorganisms with corresponding capabilities towards three organochlorine pesticides (Lindane, DDT and PCP) has been carried out.
The tolerance of plants to tested pesticides and induction degree of plant detoxification enzymes by these compounds have been used as main criteria for estimating the applicability of plants in proposed technology. Obtained results show that alfalfa, maize and soybean among tested six plant species have highest tolerance to pesticides.
As a result of screening, more than 30 strains from genera Pseudomonas have been selected. As a result of GC analysis of incubation area, 11 active cultures for investigated pesticides are carefully chosen.
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 cellulase enzymes do exist widely among microorganisms. Accordingly, the cellulose degradation using microbial cellulase to produce a low-cost substrate for ethanol production has attracted more attention. In this study, the cellulase 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 cellulase activity. The optimal temperature for its growth and cellulase production is 37°C. The optimal temperature of bacterial cellulase activity is 60°C. The cellulase enzyme from Acinetobacter sp. KKU44 is heat-tolerant enzyme. The bacterial culture of 36h. showed highest cellulase activity at 120U/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 cellulase 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.
Abstract: This article comprises detail information about L-asparaginase, encompassing topic such as various sources of L-asparaginase, mechanism and properties of L-asparaginase. Also describe the production, cultivation and purification of L-asparaginase along with information about the application of L-asparaginase. L-asparaginase catalyzes the conversion reaction to convert asparagine to aspartic acid and ammonia. Asparagine is a nutritional requirement for both normal and tumor cell. Present scenario has found that L-asparaginase has been found to be a best anti tumor or antileukemic agent. In the recent years this enzyme gained application in the field of clinical research pharmacologic and food industry. It has been characterized based on the enzyme assay principle hydrolyzing L-asparagine into L-aspartic acid and ammonia. It has been observed that eukaryotic microorganisms such as yeast and filamentous fungi have a potential for L-asparaginase production. L-asparaginase has been and is still one of the most lengthily studied therapeutic enzymes by scientist and researchers worldwide.
Abstract: Collagen was isolated from chicken feet by using papain and pepsin enzymes in acetic acid solution at 4°C for 24h with a yield of 18.16% and 22.94% by dry weight, respectively. Chemical composition and characteristics of chicken feet collagen such as amino acid composition, SDS-PAGE patterns, FTIR spectra and thermal properties were evaluated. The chicken feet collagen is rich in the amino acids glycine, glutamic acid, proline and hydroxyproline. Electrophoresis pattern demonstrated two distinct α-chains (α1 and α2) and β chain, indicating that type I collagen is a major component of chicken feet collagen. The thermal stability of collagen isolated by papain and pepsin revealed stable denaturation temperatures of 48.40 and 53.35°C, respectively. The FTIR spectra of both collagens were similar with amide regions in A, B, I, II and III. The study demonstrated that chicken feet collagen using papain isolation method is possible as commercial alternative ingredient.
Abstract: Aldehyde oxidase is molybdo-flavoenzyme involved in the oxidation of hundreds of endogenous and exogenous and N-heterocyclic compounds and environmental pollutants. Uncharged N-heterocyclic aromatic compounds such phenanthridine are commonly distributed pollutants in soil, air, sediments, surface water and groundwater, and in animal and plant tissues. Phenanthridine as uncharged N-heterocyclic aromatic compound was incubated with partially purified aldehyde oxidase from rainbow trout fish liver. Reversed-phase HLPC method was used to separate the oxidation products from phenanthridine and the metabolite was identified. The 6(5H)-phenanthridinone was identified the major metabolite by partially purified aldehyde oxidase from fish liver. Kinetic constant for the oxidation reactions were determined spectrophotometrically and showed that this substrate has a good affinity (Km = 78 ± 7.6µM) for hepatic aldehyde oxidase, will be a significant pathway. This study confirms that partially purified aldehyde oxidase from fish liver is indeed the enzyme responsible for the in vitro production 6(5H)-phenanthridinone metabolite as it is a major metabolite by mammalian aldehyde oxidase, coupled with a relatively high oxidation rate (0.77± 0.03 nmol/min/mg protein). In addition, the kinetic parameters of hepatic fish aldehyde oxidase towards the phenanthridine substrate indicate that in vitro biotransformation by hepatic fish aldehyde oxidase will be a significant pathway. This study confirms that partially purified aldehyde oxidase from fish liver is indeed the enzyme responsible for the in vitro production 6(5H)-phenanthridinone metabolite as it is a major metabolite by mammalian aldehyde oxidase.
Abstract: Altered drug binding may be an important factor in isoniazid (INH) resistance, rather than major changes in the enzyme’s activity as a catalase or peroxidase (KatG). The identification of structural or functional defects in the mutant KatGs responsible for INH resistance remains as an area to be explored. In this connection, the differences in the binding affinity between wild-type (WT) and mutants of KatG were investigated, through the generation of three mutants of KatG, Ser315Thr [S315T], Ser315Asn [S315N], Ser315Arg [S315R] and a WT [S315]) with the help of software-MODELLER. The mutants were docked with INH using the software-GOLD. The affinity is lower for WT than mutant, suggesting the tight binding of INH with the mutant protein compared to WT type. These models provide the in silico evidence for the binding interaction of KatG with INH and implicate the basis for rationalization of INH resistance in naturally occurring KatG mutant strains of Mycobacterium tuberculosis.
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.
Abstract: Conversion of lignocellulosic biomass is the basis process for production of fuels, chemicals and materials in the sustainable biorefinery industry. Saccharification of lignocellulosic biomass is an essential step which produces sugars for further conversion to target value-added products e.g. bio-ethanol, bio-plastic, g-valerolactone (GVL), 5-hydroxymethylfuroic acid (HMF), levulinic acid, etc. The goal of this work was to develop an efficient enzyme for conversion of biomass to reducing sugar based on crude fungal enzyme from Chaetomium globosum BCC5776 produced by submerged fermentation and evaluate its activity comparing to a commercial Acremonium cellulase. Five local biomasses in Thailand: rice straw, sugarcane bagasse, corncobs, corn stovers, and palm empty fruit bunches were pretreated and hydrolyzed with varying enzyme loadings. Saccharification of the biomass led to different reducing sugar levels from 115 mg/g to 720 mg/g from different types of biomass using cellulase dosage of 9 FPU/g. The reducing sugar will be further employed as sugar feedstock for production of ethanol or commodity chemicals. This work demonstrated the use of promising enzyme candidate for conversion of local lignocellulosic biomass in biorefinery industry.
Abstract: The present study is aimed at alteration of sewage sludge into stable compost product using vermicomposting of sewage sludge mixed with cattle manure and saw dust in five different proportions based on C/N ratios (C/N 15 (R1), 20 (R2), 25 (R3) and 30 (R4); and control (R5)) by employing an epigeic earthworm Eisenia fetida. Higher reductions in C/N ratio, CO2 evolution and OUR were observed in R4 demonstrated the compost stability. In addition, R4 proved to be best combination for the growth of the earthworms. In order to observe the optimal degradation, kinetics for degradation of organic matter in vermicomposting were quantitatively evaluated. An approach model was developed by assuming that composting process is carried out in a homogeneous way and the kinetics for decomposition reaction is represented by a Monod-type equation. The results exhibit comparable variations in the kinetic constants Km and K3 under varying parameters during vermicomposting process. Results suggested that higher R2 value in R4, enhanced suitability towards Lineweaver-Burke plot. R4 yields higher degradability coefficient (K) reveals that the occurrence of optimal nutrient balance, which not only enhanced the affinity of enzymes towards substrate but also improved its degradation process. Therefore, it can be proved that R4 provided to be the best feed combination for vermicomposting process as compared to other reactors.