Publications | Biotechnology and Bioengineering
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 276

World Academy of Science, Engineering and Technology

[Biotechnology and Bioengineering]

Online ISSN : 1307-6892

66 Anaerobic Treatment of Produced Water

Authors: F. C. Khong, M. H. Isa, S. R. M. Kutty, S. A. Farhan

Abstract:

An experimental study of anaerobic treatment was performed by hybrid upflow anaerobic sludge blanket (HUASB) reactor to treat produced water (PW) of an onshore crude oil terminal (COD: 1597 mg/L, NH3-N: 14.7 mg/L, phenol: 13.8 mg/L, BOD5: 862 mg/L, sodium: 6240 mg/L and chloride 9530 mg/L). The produced water with high salinity and other toxic substances will inhibit the methanogens performance if there is no adaptation on biomass before anaerobic digestion. COD removal from produced water was investigated at five different dilutions of produced water and tap water (TW) without any nutrient addition and pre-treatment. The dilution ratios were 1PW:4TW, 2PW:3TW, 3PW:2TW, 4PW:1TW and 5PW:0TW. The reactor was evaluated at mesophilic operating condition (35 ± 2 °C) at 5 days of HRT for 250 days continuous feed. The average COD removals for 1PW:4TW, 2PW:3TW, 3PW:2TW, 4PW:1TW and 5PW:0TW were found to be approximately 76.1%, 73.8%, 70.3%, 46.3% and 61.82% respectively, with final average effluent COD of 123.7 mg/L, 240 mg/L, 294 mg/L, 589 mg/L and 738 mg/L, respectively.

Keywords: Anaerobic, fixed film, hybrid UASB, produced water, inhibitor

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65 Finding Approximate Tandem Repeats with the Burrows-Wheeler Transform

Authors: Agnieszka Danek, Rafał Pokrzywa

Abstract:

Approximate tandem repeats in a genomic sequence are two or more contiguous, similar copies of a pattern of nucleotides. They are used in DNA mapping, studying molecular evolution mechanisms, forensic analysis and research in diagnosis of inherited diseases. All their functions are still investigated and not well defined, but increasing biological databases together with tools for identification of these repeats may lead to discovery of their specific role or correlation with particular features. This paper presents a new approach for finding approximate tandem repeats in a given sequence, where the similarity between consecutive repeats is measured using the Hamming distance. It is an enhancement of a method for finding exact tandem repeats in DNA sequences based on the Burrows- Wheeler transform.

Keywords: approximate tandem repeats, Burrows-Wheeler transform, Hamming distance, suffix array

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64 Exons and Introns Classification in Human and Other Organisms

Authors: Benjamin Y. M. Kwan, Jennifer Y. Y. Kwan, Hon Keung Kwan

Abstract:

In the paper, the relative performances on spectral classification of short exon and intron sequences of the human and eleven model organisms is studied. In the simulations, all combinations of sixteen one-sequence numerical representations, four threshold values, and four window lengths are considered. Sequences of 150-base length are chosen and for each organism, a total of 16,000 sequences are used for training and testing. Results indicate that an appropriate combination of one-sequence numerical representation, threshold value, and window length is essential for arriving at top spectral classification results. For fixed-length sequences, the precisions on exon and intron classification obtained for different organisms are not the same because of their genomic differences. In general, precision increases as sequence length increases.

Keywords: Exons and introns classification, Human genome, Model organism genome, Spectral analysis

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63 Chitosan Nanoparticle as a Novel Delivery System for A/H1n1 Influenza Vaccine: Safe Property and Immunogenicity in Mice

Authors: Nguyen Anh Dzung, Nguyen Thi Ngoc Hà, Dang Thi Hong Van, Nguyen Thi Lan Phuong, Nguyen Thi Nhu Quynh, Dinh Minh Hiep, Le Van Hiep

Abstract:

The aims of this paper are to study the efficacy of chitosan nanoparticles in stimulating specific antibody against A/H1N1 influenza antigen in mice. Chitosan nanoparticles (CSN) were characterized by TEM. The results showed that the average size of CSN was from 80nm to 106nm. The efficacy of A/H1N1 influenza vaccine loaded on the surface of CSN showed that loading efficiency of A/H1N1 influenza antigen on CSN was from 93.75 to 100%. Safe property of the vaccine were tested. In 10 days post vaccination, group of CSN 30 kDa and 300 kDa loaded A/H1N1 influenza antigen were the rate of immune response on mice to be 100% (9/9) higher than Al(OH)3 and other adjuvant. 100% mice in the experiment of all groups had immune response in 20 days post vaccination. The results also showed that HI titer of the group using CSN 300 kDa as an adjuvant increased significantly up to 3971 HIU, over three-fold higher than the Al(OH)3 adjuvant, chitosan (CS), and one hundredfold than the A/H1N1 antigen only. Stability of the vaccine formulation was investigated.

Keywords: Chitosan nanoparticles, A/H1N1 influenza antigen, vaccine, immunogenicity, adjuvant, antibody titer

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62 Protein Residue Contact Prediction using Support Vector Machine

Authors: Chan Weng Howe, Mohd Saberi Mohamad

Abstract:

Protein residue contact map is a compact representation of secondary structure of protein. Due to the information hold in the contact map, attentions from researchers in related field were drawn and plenty of works have been done throughout the past decade. Artificial intelligence approaches have been widely adapted in related works such as neural networks, genetic programming, and Hidden Markov model as well as support vector machine. However, the performance of the prediction was not generalized which probably depends on the data used to train and generate the prediction model. This situation shown the importance of the features or information used in affecting the prediction performance. In this research, support vector machine was used to predict protein residue contact map on different combination of features in order to show and analyze the effectiveness of the features.

Keywords: contact map, protein residue contact, support vector machine, protein structure prediction

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61 Cloning and Expression of D-Threonine Aldolase from Ensifer arboris NBRC100383

Authors: Sang-Ho Baik

Abstract:

D-erythro-cyclohexylserine (D chiral unnatural β-hydroxy amino acid expected for the synthesis of drug for AIDS treatment. To develop a continuous bioconversion system with whole cell biocatalyst of D-threonine aldolase (D genes for the D-erythro-CHS production, D-threonine aldolase gene was amplified from Ensifer arboris 100383 by direct PCR amplication using two degenerated oligonucleotide primers designed based on genomic sequence of Shinorhizobium meliloti Sequence analysis of the cloned DNA fragment revealed one open-reading frame of 1059 bp and 386 amino acids. This putative D-TA gene was cloned into NdeI and EcoRI (pEnsi His-tag sequence or BamHI (pEnsi-DTA[2]) sequence of the pET21(a) vector. The expression level of the cloned gene was extremely overexpressed by E. coli BL21(DE3) transformed with pEnsi-DTA[1] compared to E. coli BL21(DE3) transformed with pEnsi-DTA[2]. When the cells expressing the wild used for D-TA enzyme activity, 12 mM glycine was successfully detected in HPLC analysis. Moreover, the whole cells harbouring the recombinant D-TA was able to synthesize D-erythro of 0.6 mg/ml in a batch reaction.

Keywords: About four key words or phrases in alphabetical order, separated by commas.

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60 Effect of Crude oil Intoxication on Antioxidant and Marker Enzymes of Tissue Damage in Liver of Rat

Authors: K. Mahmoud, T. Shalahmetova, Sh. Deraz

Abstract:

The objective of the present study was to examine the dose-response relationships between antioxidant parameters and liver contaminant levels of Kazakhstan light crude oil (KLCO) in albino rats. The animals were repeatedly exposed, by intraperitoneal injection, to low dosages (0.5–1.5 ml/kg) of KLCO. Rats exposed to these doses levels did not show any apparent symptoms of intoxication. Serum aminotransferases increased significantly (p<0.05) while after 8-d recovered to control level. hepatic superoxide dismutase (SOD) activity significantly increased after 1 and 3-d except with 0.5 ml while with increased exposure time it deceased insignificantly after 5-d and significantly after 8-d. Dose and time-dependent increases were observed in the levels of hepatic glutathione-S-transeferase (GST) and conjugated diene (CD) while malondialdehyde (MDA) was evolved after 8 with the dose of 1.5 ml. the data obtained indicate impaired liver function in the rats intoxicated with KLCO as manifested by elevated liver enzymes.

Keywords: Crude oil; superoxide dismutase, glutathione-Stranseferase, malondialdehyde, conjugated diene, aminotransferases.

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59 Characterization of the O.ul-mS952 Intron:A Potential Molecular Marker to Distinguish Between Ophiostoma Ulmi and Ophiostoma Novo-Ulmi Subsp. Americana

Authors: Mohamed Hafez, Georg Hausner

Abstract:

The full length mitochondrial small subunit ribosomal (mt-rns) gene has been characterized for Ophiostoma novo-ulmi subspecies americana. The gene was also characterized for Ophiostoma ulmi and a group II intron was noted in the mt-rns gene of O. ulmi. The insertion in the mt-rns gene is at position S952 and it is a group IIB1 intron that encodes a double motif LAGLIDADG homing endonuclease from an open reading frame located within a loop of domain III. Secondary structure models for the mt-rns RNA of O. novo-ulmi subsp. americana and O. ulmi were generated to place the intron within the context of the ribosomal RNA. The in vivo splicing of the O.ul-mS952 group II intron was confirmed with reverse transcription-PCR. A survey of 182 strains of Dutch Elm Diseases causing agents showed that the mS952 intron was absent in what is considered to be the more aggressive species O. novo-ulmi but present in strains of the less aggressive O. ulmi. This observation suggests that the O.ul-mS952 intron can be used as a PCR-based molecular marker to discriminate between O. ulmi and O. novo-ulmi subsp. americana.

Keywords: Dutch Elm Disease, group II introns, mtDNA, species identification

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58 Antioxidant Responses to Different Exposure Regimes of Kazakhstan Light Crude Oil in Livers of Male Albino Rats

Authors: K. Mahmoud, T. Shalahmetova, B. Umbayev, Sh. Deraz

Abstract:

Biochemical investigations were carried out to assess the effect of different exposure regimes of Kazakhstan crude oil (KCO) on hepatic antioxidant defense system in albino rats. Contaminants were delivered under two different dosing regimes, with all treatments receiving the same total contaminant load by the end of the exposure period. Rats in regime A injected with KCO once at a dose of 6 ml/kg bw while in regime B injected multiply at a dose of 1.5 ml/kg bw on day 1, 3, 5 and 8. Antioxidant biomarkers were measured in hepatic tissue after 1, 3, 5 and 8 days. Significant induction was observed in serum aminotransferases (ALT, AST) (p<0.01) and hepatic Glutathione-S-transferase (GST) (p<0.05) in the two exposure regimes, with the majority of significant induction occurring in regime A. Superoxide dismutase (SOD) increased 1-d after injection (p<0.01) but the increase was reduced time dependently thereafter while after 8-d induced again (p<0.01). Malondialdehyde (MDA) significantly induced after 3 and 5-d (p<0.05) in regime A while in regime B was not changed significantly (p>0.05) at short time after exposure. However, there was significant increase after 8-d (p<0.01). Histological examination indicates that crude oil induced pathologic changes from inflammatory cells infiltration to hemorrhage and necrosis of hepatocytes. Acute exposure to crude oil adversely affect hepatic cell so human must avoid such exposure.

Keywords: Kazakhstan crude oil, Antioxidant biomarkers, Histological examination, Dose regime, Rats

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57 Protein Production by Bacillus Subtilis Atcc 21332 in the Presence of Cymbopogon Essential Oils

Authors: Hanina M. N., Hairul Shahril M., Mohd Fazrullah Innsan M. F., Ismatul Nurul Asyikin I., Abdul Jalil A. K, Salina M. R., Ahmad I.B.

Abstract:

Proteins levels produced by bacteria may be increased in stressful surroundings, such as in the presence of antibiotics. It appears that many antimicrobial agents or antibiotics, when used at low concentrations, have in common the ability to activate or repress gene transcription, which is distinct from their inhibitory effect. There have been comparatively few studies on the potential of antibiotics or natural compounds in nature as a specific chemical signal that can trigger a variety of biological functions. Therefore, this study was focusing on the effect of essential oils from Cymbopogon flexuosus and C. nardus in regulating proteins production by Bacillus subtilis ATCC 21332. The Minimum Inhibition Concentrations (MICs) of both essential oils on B. subtilis were determined by using microdilution assay, resulting 0.2% and 1.56% for each C. flexuosus and C. nardus subsequently. The bacteria were further exposed to each essential oils at concentration of 0.01XMIC for 2 days. The proteins were then isolated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Protein profile showed that a band with approximate size of 250 kD was appeared for the treated bacteria with essential oils. Thus, Bacillus subtilis ATCC 21332 in stressful condition with the presence of essential oils at low concentration could induce the protein production.

Keywords: Bacillus subtilis ATCC 21332, Cymbopogon essential oils, protein

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56 Characteristics of Intronic and Intergenic Human miRNAs and Features of their Interaction with mRNA

Authors: Assel S. Issabekova, Olga A. Berillo, Vladimir A. Khailenko, Shara A. Atambayeva, Mireille Regnier, Anatoly T. Ivachshenko

Abstract:

Regulatory relationships of 686 intronic miRNA and 784 intergenic miRNAs with mRNAs of 51 intronic miRNA coding genes were established. Interaction features of studied miRNAs with 5'UTR, CDS and 3'UTR of mRNA of each gene were revealed. Functional regions of mRNA were shown to be significantly heterogenous according to the number of binding sites of miRNA and to the location density of these sites.

Keywords: 5'UTR, 3'UTR, CDS, miRNA, target mRNA

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55 Construction of cDNALibrary and EST Analysis of Tenebriomolitorlarvae

Authors: JiEun Jeong, Se-Won Kang, Hee-Ju Hwang, Sung-Hwa Chae, Sang-Haeng Choi, Hong-SeogPark, YeonSoo Han, Bok-Reul Lee, Dae-Hyun Seog, Yong Seok Lee

Abstract:

Tofurther advance research on immune-related genes from T. molitor, we constructed acDNA library and analyzed expressed sequence taq (EST) sequences from 1,056 clones. After removing vector sequence and quality checkingthrough thePhred program (trim_alt 0.05 (P-score>20), 1039 sequences were generated. The average length of insert was 792 bp. In addition, we identified 162 clusters, 167 contigs and 391 contigs after clustering and assembling process using a TGICL package. EST sequences were searchedagainst NCBI nr database by local BLAST (blastx, EKeywords: EST, Innate immunity, Tenebriomolitor

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54 Improved Technique of Non-viral Gene Delivery into Cancer Cells

Authors: D. Vainauska, S. Kozireva, A. Karpovs, M. Chistyakovs, M. Baryshev

Abstract:

Liposomal magnetofection is a simple, highly efficient technology for cell transfection, demonstrating better outcome than a number of other common gene delivery methods. However, aggregate complexes distribution over the cell surface is non-uniform due to the gradient of the permanent magnetic field. The aim of this study was to estimate the efficiency of liposomal magnetofection for prostate carcinoma PC3 cell line using newly designed device, “DynaFECTOR", ensuring magnetofection in a dynamic gradient magnetic field. Liposomal magnetofection in a dynamic gradient magnetic field demonstrated the highest transfection efficiency for PC3 cells – it increased for 21% in comparison with liposomal magnetofection and for 42% in comparison with lipofection alone. The optimal incubation time under dynamic magnetic field for PC3 cell line was 5 minutes and the optimal rotation frequency of magnets – 5 rpm. The new approach also revealed lower cytotoxic effect to cells than liposomal magnetofection.

Keywords: Dynamic gradient magnetic field, gene delivery, liposomal magnetofection, prostate cancer cell line

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53 A New Edit Distance Method for Finding Similarity in Dna Sequence

Authors: Patsaraporn Somboonsak, Mud-Armeen Munlin

Abstract:

The P-Bigram method is a string comparison methods base on an internal two characters-based similarity measure. The edit distance between two strings is the minimal number of elementary editing operations required to transform one string into the other. The elementary editing operations include deletion, insertion, substitution two characters. In this paper, we address the P-Bigram method to sole the similarity problem in DNA sequence. This method provided an efficient algorithm that locates all minimum operation in a string. We have been implemented algorithm and found that our program calculated that smaller distance than one string. We develop PBigram edit distance and show that edit distance or the similarity and implementation using dynamic programming. The performance of the proposed approach is evaluated using number edit and percentage similarity measures.

Keywords: Edit distance, String Matching, String Similarity

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52 Oleate Induces Apoptosis in 3T3-L1 Adipocytes

Authors: A. Rohana, A. M., Fadzilah Adibah, M. S. Muhammad Roji

Abstract:

Oleic acid (C18:1) play an important role in proliferation of fat cells. In this study, the effect of oleate on cells viability in 3T3-L1 cells (fat cells) was investigated. The 3T3-L1 cells were treated with various concentrations of oleate in the presence of 23 mM glucose. Oleate was added to adipogenic media (day 0) to investigate the influence of oleate on proliferation of postconfluent preadipocytes after 24 h induction. 0.1 mM oleate promoted cell division by increasing 33.9% number of cells from basal control in postconfluent preadipocytes. However, there were no significantly different in cells viability with control cells when oleate concentrations were increased up to 0.5 mM. When added to differentiated adipocytes (day 12) for 48 h, the number of cells decreased as oleate concentrations increased. 92.7% of cells lost demonstrated apoptosis and necrosis after 48 h with 0.5 mM oleate. The fluorochrome staining was examined under fluorescence microscopy using acridine orange and ethidium bromide double staining. Furthermore, the presence of high lactate (60.6% increased from basal control) released into plasma has shown the direct cytotoxicity of 0.5 mM oleate on adipocytes.

Keywords: adipocytes, apoptosis, oleate, postconfluentpreadipocytes

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51 Product Development and Derivatives Exploration by using Photosynthetic Bacteria

Authors: Yi-Fang Hung, Jinn-Tsyy Lai

Abstract:

Lycopene, which can be extracted from plants and is very popular for fruit intake, is restricted for healthy food development due to its high price. On the other hand, it will get great safety concerns, especially in the food or cosmetic application, if the raw material of lycopene is produced by chemical synthesis. In this project, we provide a key technology to bridge the limitation as mentioned above. Based on the abundant bioresources of BCRC (Bioresource Collection and Research Center, Taiwan), a promising lycopene output will be anticipated by the introduction of fermentation technology along with industry-related core energy. Our results showed that addition of tween 80(0.2%) and span 20 produced higher amount of lycopene. And piperidine, when was added at 48hr to the cultivation medium, could promote lycopene excretion effectively also.

Keywords: photosynthetic bacteria, lycopene, tween80, Piperidine

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50 Full-genomic Network Inference for Non-model organisms: A Case Study for the Fungal Pathogen Candida albicans

Authors: Jörg Linde, Ekaterina Buyko, Robert Altwasser, Udo Hahn, Reinhard Guthke

Abstract:

Reverse engineering of full-genomic interaction networks based on compendia of expression data has been successfully applied for a number of model organisms. This study adapts these approaches for an important non-model organism: The major human fungal pathogen Candida albicans. During the infection process, the pathogen can adapt to a wide range of environmental niches and reversibly changes its growth form. Given the importance of these processes, it is important to know how they are regulated. This study presents a reverse engineering strategy able to infer fullgenomic interaction networks for C. albicans based on a linear regression, utilizing the sparseness criterion (LASSO). To overcome the limited amount of expression data and small number of known interactions, we utilize different prior-knowledge sources guiding the network inference to a knowledge driven solution. Since, no database of known interactions for C. albicans exists, we use a textmining system which utilizes full-text research papers to identify known regulatory interactions. By comparing with these known regulatory interactions, we find an optimal value for global modelling parameters weighting the influence of the sparseness criterion and the prior-knowledge. Furthermore, we show that soft integration of prior-knowledge additionally improves the performance. Finally, we compare the performance of our approach to state of the art network inference approaches.

Keywords: Pathogen, network inference, text-mining, Candida albicans, LASSO, mutual information, reverse engineering, linear regression, modelling.

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49 A Stochastic Approach of Mitochondrial Dynamics

Authors: Athanasios T. Alexiou, Maria M. Psiha, John A. Rekkas, Panayiotis M. Vlamos

Abstract:

Mitochondria are dynamic organelles, capable to interact with each other. While the number of mitochondria in a cell varies, their quality and functionality depends on the operation of fusion, fission, motility and mitophagy. Nowadays, several researches declare as an important factor in neurogenerative diseases the disruptions in the regulation of mitochondrial dynamics. In this paper a stochastic model in BioAmbients calculus is presented, concerning mitochondrial fusion and its distribution in the renewal of mitochondrial population in a cell. This model describes the successive and dependent stages of protein synthesis, protein-s activation and merging of two independent mitochondria.

Keywords: Mitochondrial Dynamics, P-Calculus, StochasticModeling.

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48 Using Malolactic Fermentation with Acid- And Ethanol- Adapted Oenococcus Oeni Strain to Improve the Quality of Wine from Champs Bourcin Grape in Sapa - Lao Cai

Authors: Pham Thu Thuy, Nguyen Lan Huong, Chu Ky Son

Abstract:

Champs Bourcin black grape originated from Aquitaine, France and planted in Sapa, Lao cai provice, exhibited high total acidity (11.72 g/L). After 9 days of alcoholic fermentation at 25oC using Saccharomyces cerevisiae UP3OY5 strain, the ethanol concentration of wine was 11.5% v/v, however the sharp sour taste of wine has been found. The malolactic fermentation (MLF) was carried out by Oenococcus oeni ATCCBAA-1163 strain which had been preadapted to acid (pH 3-4) and ethanol (8-12%v/v) conditions. We obtained the highest vivability (83.2%) upon malolactic fermentation after 5 days at 22oC with early stationary phase O. oeni cells preadapted to pH 3.5 and 8% v/v ethanol in MRS medium. The malic acid content in wine was decreased from 5.82 g/L to 0.02 g/L after MLF (21 days at 22oC). The sensory quality of wine was significantly improved.

Keywords: Champs Bourcin grape, malolactic fermentation, pre-adaptation, Oenococcus oeni

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47 Possible Protective Effect of Kombucha Tea Ferment on Cadmium Chloride Induced Liver and Kidney Damage in Irradiated Rats

Authors: Nashwa Kamel Ibrahim

Abstract:

Kombucha Tea Ferment (KT), was given to male albino rats, (1ml/Kg of body weight), via gavages, during 2 weeks before intraperitoneal administration of 3.5 mg/Kg body weight CdCl2 and/or whole body γ-irradiation with 4Gy, and during 4 weeks after each treatment. Hepatic and nephritic pathological changes included significant increases of serum alanine transaminase (ALT), aspartate transaminase (AST), and alkaline phosphatase (ALP) activities, and creatinine and urea contents with significant decrease in serum total antioxidant capacity (TAC). Increase in oxidative stress markers in liver and kidney tissues expressed by significant increase in malondialdehyde (MDA) and nitric oxide (NO) contents associated to significant depletion in superoxide dismutase (SOD) and catalase (CAT) activities, and reduced glutathione (GSH) content were recorded. KT administration results in recovery of all the pathological changes. It could be concluded that KT might protect liver and kidney from oxidative damage induced by exposure to cadmium and/ or γ-irradiation.

Keywords: Cadmium, Kombucha, radiation, rats

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46 Micro-aerobic, Anaerobic and Two-stage Condition for Ethanol Production by enterobacter aerogenes from Biodiesel-derived Crude Glycerol

Authors: Kanokrat Saisaard, Irini Angelidaki, Poonsuk Prasertsan

Abstract:

The microbial production of ethanol from biodiesel¬derived crude glycerol by Enterobacter aerogenes TISTR1468, under micro-aerobic and anaerobic conditions, was investigated. The experimental results showed that micro-aerobic conditions were more favorable for cellular growth (4.0 g/L DCW), ethanol production (20.7 g/L) as well as the ethanol yield (0.47 g/g glycerol) than anaerobic conditions (1.2 g/L DCW, 6.3 g/L ethanol and 0.72 g/g glycerol, respectively). Crude glycerol (100 g/L) was consumed completely with the rate of 1.80 g/L/h. Two-stage fermentation (combination of micro-aerobic and anaerobic condition) exhibited higher ethanol production (24.5 g/L) than using one-stage fermentation (either micro-aerobic or anaerobic condition. The two- stage configuration, exhibited slightly higher crude glycerol consumption rate (1.81 g/L/h), as well as ethanol yield (0.56 g/g) than the one-stage configuration. Therefore, two-stage process was selected for ethanol production from E. aerogenes TISTR1468 in scale-up studies.

Keywords: crude glycerol, ethanol, micro-aerobic, two-stage, Enterobacter aerogenes

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45 High-Intensity Nanosecond Pulsed Electric Field effects on Early Physiological Development in Arabidopsis thaliana

Authors: Wisuwat Songnuan, Phumin Kirawanich

Abstract:

The influences of pulsed electric fields on early physiological development in Arabidopsis thaliana were studied. Inside a 4-mm electroporation cuvette, pre-germination seeds were subjected to high-intensity, nanosecond electrical pulses generated using laboratory-assembled pulsed electric field system. The field strength was varied from 5 to 20 kV.cm-1 and the pulse width and the pulse number were maintained at 10 ns and 100, respectively, corresponding to the specific treatment energy from 300 J.kg-1 to 4.5 kJ.kg-1. Statistical analyses on the average leaf area 5 and 15 days following pulsed electric field treatment showed that the effects appear significant the second week after treatments with a maximum increase of 80% compared to the control (P < 0.01).

Keywords: Arabidopsis thaliana, full-wave analysis, leaf area, high-intensity nanosecond pulsed electric fields

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44 Microalgal Lipid Production by Microalgae Chlorella sp. KKU-S2

Authors: Ratanaporn Leesing, Supaporn Kookkhunthod, Ngarmnit Nontaso

Abstract:

The objective of this work is to produce heterotrophic microalgal lipid in flask-batch fermentation. Chlorella sp. KKU-S2 supported maximum values of 0.374 g/L/d, 0.478 g lipid/g cells, and 0.112 g/L/d for volumetric lipid production rate, and specific yield of lipid, and specific rate of lipid production, respectively when culture was performed on BG-11 medium supplemented with 50g/L glucose. Among the carbon sources tested, maximum cell yield coefficient (YX/S, g/L), maximum specific yield of lipid (YP/X, g lipid/g cells) and volumetric lipid production rate (QP, g/L/d) were found of 0.728, 0.237, and 0.619, respectively, using sugarcane molasses as carbon source. The main components of fatty acid from extracted lipid were palmitic acid, stearic acid, oleic acid and linoleic acid which similar to vegetable oils and suitable for biodiesel production.

Keywords: Microalgal lipid, Chlorella sp. KKU-S2, kineticparameters, biodiesel.

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43 Neutralization of Alkaline Waste-Waters using a Blend of Microorganisms

Authors: Rita Kumar, Alka Sharma, Purnima Dhall, Niha M. Kulshreshtha, Anil Kumar

Abstract:

The efficient operation of any biological treatment process requires pre-treatment of incompatible pollutants such as acids, bases, oil, toxic substances, etc. which hamper the treatment of other major components which are otherwise degradable. The pre-treatment of alkaline waste-waters, generated from various industries like textile, paper & pulp, potato-processing industries, etc., having a pH of 10 or higher, is essential. The pre-treatment, i.e., neutralization of such alkaline waste-waters can be achieved by chemical as well as biological means. However, the biological pretreatment offers better package over the chemical means by being safe and economical. The biological pre-treatment can be accomplished by using a blend of microorganisms able to withstand such harsh alkaline conditions. In the present study, for the proper pre-treatment of alkaline waste-waters, a package of alkalophilic bacteria is formulated to neutralise the alkaline pH of the industrial waste-waters. The developed microbial package is cost-effective as well as environmental friendly.

Keywords: alkaline, alkalophilic bacteria, biological, pollutants, textile.

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42 Microbial Oil Production by Isolated Oleaginous Yeast Torulaspora globosa YU5/2

Authors: Ratanaporn Leesing, Ratanaporn Baojungharn

Abstract:

Microbial oil was produced by soil isolated oleaginous yeast YU5/2 in flask-batch fermentation. The yeast was identified by molecular genetics technique based on sequence analysis of the variable D1/D2 domain of the large subunit (26S) ribosomal DNA and it was identified as Torulaspora globosa. T. globosa YU5/2 supported maximum values of 0.520 g/L/d, 0.472 g lipid/g cells, 4.16 g/L, and 0.156 g/L/d for volumetric lipid production rate, and specific yield of lipid, lipid concentration, and specific rate of lipid production respectively, when culture was performed in nitrogen-limiting medium supplemented with 80g/L glucose. Among the carbon sources tested, maximum cell yield coefficient (YX/S, g/L), maximum specific yield of lipid (YP/X, g lipid/g cells) and volumetric lipid production rate (QP, g/L/d) were found of 0.728, 0.237, and 0.619, respectively, using sweet potato tubers hydrolysates as carbon source.

Keywords: Microbial oil, oleaginous yeast, Torulasporaglobosa YU5/2, sweet potato tubers, kinetic parameters.

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41 Construction of Recombinant E.coli Expressing Fusion Protein to Produce 1,3-Propanediol

Authors: Rosarin Rujananon, Poonsuk Prasertsan, Amornrat Phongdara, Tanate Panrat, Jibin Sun, Sugima Rappert, An-Ping Zeng

Abstract:

In this study, a synthetic pathway was created by assembling genes from Clostridium butyricum and Escherichia coli in different combinations. Among the genes were dhaB1 and dhaB2 from C. butyricum VPI1718 coding for glycerol dehydratase (GDHt) and its activator (GDHtAc), respectively, involved in the conversion of glycerol to 3-hydroxypropionaldehyde (3-HPA). The yqhD gene from E.coli BL21 was also included which codes for an NADPHdependent 1,3-propanediol oxidoreductase isoenzyme (PDORI) reducing 3-HPA to 1,3-propanediol (1,3-PD). Molecular modeling analysis indicated that the conformation of fusion protein of YQHD and DHAB1 was favorable for direct molecular channeling of the intermediate 3-HPA. According to the simulation results, the yqhD and dhaB1 gene were assembled in the upstream of dhaB2 to express a fusion protein, yielding the recombinant strain E. coliBL21 (DE3)//pET22b+::yqhD-dhaB1_dhaB2 (strain BP41Y3). Strain BP41Y3 gave 10-fold higher 1,3-PD concentration than E. coliBL21 (DE3)//pET22b+::yqhD-dhaB1_dhaB2 (strain BP31Y2) expressing the recombinant enzymes simultaneously but in a non-fusion mode. This is the first report using a gene fusion approach to enhance the biological conversion of glycerol to the value added compound 1,3- PD.

Keywords: Recombinant E.coli, 1, 3-propanediol, glycerol, fusion protein.

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40 Detection of Oxidative Stress Induced by Mobile Phone Radiation in Tissues of Mice using 8-Oxo-7, 8-Dihydro-2'-Deoxyguanosine as a Biomarker

Authors: Ahmad M. Khalil, Ahmad M. Alshamali, Marwan H. Gagaa

Abstract:

We investigated oxidative DNA damage caused by radio frequency radiation using 8-oxo-7, 8-dihydro-2'- deoxyguanosine (8-oxodG) generated in mice tissues after exposure to 900 MHz mobile phone radio frequency in three independent experiments. The RF was generated by a Global System for Mobile Communication (GSM) signal generator. The radio frequency field was adjusted to 25 V/m. The whole body specific absorption rate (SAR) was 1.0 W/kg. Animals were exposed to this field for 30 min daily for 30 days. 24 h post-exposure, blood serum, brain and spleen were removed and DNA was isolated. Enzyme-linked immunosorbent assay (ELISA) was used to measure 8-oxodG concentration. All animals survived the whole experimental period. The body weight of animals did not change significantly at the end of the experiment. No statistically significant differences observed in the levels of oxidative stress. Our results are not in favor of the hypothesis that 900 MHz RF induces oxidative damage.

Keywords: Mice, Mobile phone radiation, oxidative stress, 8-oxo-7, 8-dihydro-2'-deoxyguanosine

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39 Interactions between Cells and Nanoscale Surfaces of Oxidized Silicon Substrates

Authors: Chung-Yao Yang, Lin-Ya Huang, Tang-Long Shen, J. Andrew Yeh

Abstract:

The importance for manipulating an incorporated scaffold and directing cell behaviors is well appreciated for tissue engineering. Here, we developed newly nano-topographic oxidized silicon nanosponges capable of being various chemical modifications to provide much insight into the fundamental biology of how cells interact with their surrounding environment in vitro. A wet etching technique is exerted to allow us fabricated the silicon nanosponges in a high-throughput manner. Furthermore, various organo-silane chemicals enabled self-assembled on the surfaces by vapor deposition. We have found that Chinese hamster ovary (CHO) cells displayed certain distinguishable morphogenesis, adherent responses, and biochemical properties while cultured on these chemical modified nano-topographic structures in compared with the planar oxidized silicon counterparts, indicating that cell behaviors can be influenced by certain physical characteristic derived from nano-topography in addition to the hydrophobicity of contact surfaces crucial for cell adhesion and spreading. Of particular, there were predominant nano-actin punches and slender protrusions formed while cells were cultured on the nano-topographic structures. This study shed potential applications of these nano-topographic biomaterials for controlling cell development in tissue engineering or basic cell biology research.

Keywords: Nanosponge, Cell adhesion, Cell morphology

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38 A New Approach In Protein Folding Studies Revealed The Potential Site For Nucleation Center

Authors: Nurul Bahiyah Ahmad Khairudin, Habibah A Wahab

Abstract:

A new approach to predict the 3D structures of proteins by combining the knowledge-based method and Molecular Dynamics Simulation is presented on the chicken villin headpiece subdomain (HP-36). Comparative modeling is employed as the knowledge-based method to predict the core region (Ala9-Asn28) of the protein while the remaining residues are built as extended regions (Met1-Lys8; Leu29-Phe36) which then further refined using Molecular Dynamics Simulation for 120 ns. Since the core region is built based on a high sequence identity to the template (65%) resulting in RMSD of 1.39 Å from the native, it is believed that this well-developed core region can act as a 'nucleation center' for subsequent rapid downhill folding. Results also demonstrate that the formation of the non-native contact which tends to hamper folding rate can be avoided. The best 3D model that exhibits most of the native characteristics is identified using clustering method which then further ranked based on the conformational free energies. It is found that the backbone RMSD of the best model compared to the NMR-MDavg is 1.01 Å and 3.53 Å, for the core region and the complete protein, respectively. In addition to this, the conformational free energy of the best model is lower by 5.85 kcal/mol as compared to the NMR-MDavg. This structure prediction protocol is shown to be effective in predicting the 3D structure of small globular protein with a considerable accuracy in much shorter time compared to the conventional Molecular Dynamics simulation alone.

Keywords: 3D model, Chicken villin headpiece subdomain, Molecular dynamic simulation NMR-MDavg, RMSD.

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37 Hydrolysis of Hull-Less Pumpkin Oil Cake Protein Isolate by Pepsin

Authors: Ivan Živanović, Žužana Vaštag, Senka Popović, Ljiljana Popović, Draginja Peričin

Abstract:

The present work represents an investigation of the hydrolysis of hull-less pumpkin (Cucurbita Pepo L.) oil cake protein isolate (PuOC PI) by pepsin. To examine the effectiveness and suitability of pepsin towards PuOC PI the kinetic parameters for pepsin on PuOC PI were determined and then, the hydrolysis process was studied using Response Surface Methodology (RSM). The hydrolysis was carried out at temperature of 30°C and pH 3.00. Time and initial enzyme/substrate ratio (E/S) at three levels were selected as the independent parameters. The degree of hydrolysis, DH, was mesuared after 20, 30 and 40 minutes, at initial E/S of 0.7, 1 and 1.3 mA/mg proteins. Since the proposed second-order polynomial model showed good fit with the experimental data (R2 = 0.9822), the obtained mathematical model could be used for monitoring the hydrolysis of PuOC PI by pepsin, under studied experimental conditions, varying the time and initial E/S. To achieve the highest value of DH (39.13 %), the obtained optimum conditions for time and initial E/S were 30 min and 1.024 mA/mg proteins.

Keywords: Enzymatic hydrolysis, Pepsin, Pumpkin (CucurbitaPepo L.) oil cake protein isolate, Response surface methodology.

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