Day 3 :
- Food Biotechnology, Agricultural Biotechnology, Biotechnology and its Applications, Environmental Biotechnology
Location: Sausalito
Session Introduction
Akhileshwari Nath
S S Hospital and Research Institute, India
Title: Molecular profiling of testis in arsenic induced mice
Biography:
Akhileshwari Nath is recipient of gold medal during B.Sc. Hons and research work associated gold medals, Common Wealth Exchange fellowship and other awards for her research work. She has completed her Ph.D. in 1974 from Patna University, Patna, Bihar (India) and did postdoctoral training on Electron Microscopy at Minneapolis, Minnesota (USA) in 1976. She is retired Professor and Head of the Dept. of Zoology, from Patna University, Patna, Bihar (India). She was the Head of the Research Centre, Mahavir Cancer Institute & Research Centre (a charitable cancer hospital) Patna, Bihar (India) till January 2015. Presently she is Head of Research Institute, S. S. Hospital and Research Institute, Kankarbagh, Patna. She has published more than 130 papers in reputed journals and completed 5 major research projects funded by Govt. of India.
Abstract:
Arsenic is a potent environmental toxicant and affects biological system through food chain causing toxicity and disturbs different signalling pathways, thus suppresses immune system and finally causing various diseases.
In previous study, extensive survey work has been made in arsenic hit area and drinking water and blood samples were collected. Tissue samples have been collected from cancer patients at S. S .Hospital and Research Institute. After the confirmation of significant high level of arsenic in drinking water, blood and tissue samples, present study was undertaken.
Present study was undertaken to observe the effect of arsenic in testicular cells in mice model and its effect on testicular gene expression.
Sodium arsenite was administered into Swiss albino mice as 2mg/kg body wt. for the different durations. Estimation of arsenic was done by atomic absorption spectrophotometer. TUNEL assay was done to observe the DNA damage and microarray analysis was performed to observe the mRNA expression profile in sodium arsenite administered mice model.
High accumulation of arsenic was found in testes of Swiss albino mice. Significant DNA damage was observed in arsenic administered testicular cells of Swiss albino mice. Further, mRNA of few genes shows their altered expression.
In the present study, it can be concluded that arsenic affects testicular cells leading to DNA damage and alter testicular gene expression. Thus, our results suggest that mice with high accumulation of arsenic shows altered gene expression.
Jong H Kim
United States Department of Agriculture-Agricultural Research Service, USA
Title: Chemobiological approaches for enhancing the efficacy of antifungal intervention
Biography:
Jong H. Kim is a researcher in the Foodborne Toxin Detection and Prevention Research Unit, Western Regional Research Center, US Department of Agriculture, Albany, California. His research focuses on the development of intervention strategies for control of pathogenic fungi. He provides chemo-biological expertise, particularly in the identification of cellular targets and compound interaction, and participates in resistance management in collaboration with industry and academia.
Abstract:
Control of fungal pathogens, such as causative agents for aspergillosis, candidiasis, cryptococcosis or producers of mycotoxins, is problematic since effective antifungal agents are often very limited. Also, the expansion of fungal resistance to conventional drugs or fungicides is a global health or food safety/security issue. Therefore, there is persistent need to improve the drug efficacy or to develop new intervention strategies. Fungal drug resistance frequently involves mutations caused by environmental stressors. In fungi, stress signals resulting from oxidative or cell wall stress are integrated into mitogen-activated protein kinase (MAPK) systems that regulate defense genes countering the stress. Of note, mutations in MAPK signaling system could result in tolerance to antifungal agents. Many natural compounds are promising antifungals or leads due to their ability to disrupt fungal defense systems, such as antioxidant pathway. Natural compounds could also serve as chemical probes for identifying new antifungal targets. To enhance drug susceptibility of fungi, the model yeast Saccharomyces cerevisiae was used as a tool for identifying cellular targets of natural compounds, where targeting vulnerable components such as antioxidant system effectively disrupted pathogen growth, overcame antifungal tolerance or inhibited mycotoxin production. Finally, chemo-biological approaches enabled the development of novel antifungal chemosensitization, which significantly improved the drug susceptibility of fungal pathogens.
Lmar Babrak
United States Department of Agriculture-Agricultural Research Service, USA
Title: Antibody engineering from hybridoma-derived monoclonal antibodies
Biography:
Dr. Lmar Babrak is currently a post-doctoral research scientist in the laboratory of Dr. Robert Hnasko at the Agricultural Research Service located in Albany, CA. Her research has focused on the development of immunoassays used for the detection of disease causing pathogens, toxins and other agricultural contaminants. She completed her PhD in Microbiology in 2015 from Oregon State University.
Abstract:
Antibody engineering requires the identification of antigen binding domains (variable regions; VRs) unique to each antibody. This determination can be achieved by sequence analysis of the antibody transcript obtained from the hybridoma; as each clonal hybridoma cell line produces in principle a single antigen specific monoclonal antibody (MAb). However, the polyploidy nature of hybridoma cells often results in the added expression of aberrant immunoglobulin-like transcripts or even production of nondescript antibodies. The occurrence of these transcripts confounds identification of the VRs of immunoglobulin heavy and light chains that correspond to the antigen specific antibody. It is the VRs that define the unique antigen binding properties and proper sequence identification is essential for functional performance of a recombinant engineered antibody. To address this problem, we have: (1) identified and complied a database of aberrant Ig-like transcripts found in myeloma cell lines (SP2/0-Ag14 and P3X62A8U.1) frequently used in the generation of hybridomas; and (2) developed a PCR-based method for the selective amplification of heavy and light chain VRs from a given antigen specific immunoglobulin isotype combined with molecular cloning and DNA based sequence analysis. These methods should increase the certainty regarding the VR sequence structure when evaluating the functional performance of a recombinant antibody. This work serves to facilitate antibody engineering applications with broad interest to biotechnology and pharmaceutical industries.
Nivedita Lal
Jamia Hamdard, India
Title: Deep sequencing of root endophyte Piriformospora indica grown under salt stress
Biography:
I have completed my Ph.D. in “Plant Molecular Biology” at the Jawaharlal Nehru University, New Delhi, India. Currently I am working on my DST young scientist project (plant-P.indica interaction under salt stress) as a PI (postdoctoral fellow) at Jaima Hamdard, New Delhi. I have participitated in various national and international conferences in order to improve my knowledge and skills. I am interested in continuing my research career in plant-microbe interaction where I could employ my skills to understand how the plant-microbe relation regulates plant development and defense response against various stresses.
Abstract:
Background: Piriformospora indica , a filamentous fungus of the order Sebacinales, is able to make symbiotic interaction with root of different plant species and provides better growth and higher yield to the host plant as well as resistance against biotic and abiotic stresses. High soil salinity, excess of NaCl, is one of the important environmental factors that limits distribution and productivity of major crops. The need to produce crops with enhanced tolerance to salt stress has been the stimulus for research. P. indica-mediated salt tolerance mechanism was found to be linked strongly with increase in antioxidants under salt stress in barley which attenuates the NaCl-induced lipid peroxidation, metabolic heat efflux, and fatty acid desaturation in barley leaves. Salt stress studies have indicated promising effect of P. indica in barley. Therefore, it is vital to isolate and functionally characterize salinity stress-related genes to elucidate the mechanisms underlying halotolerance and develop salinity stress-tolerant plants.
Observations: We have compared the transcriptome of P. indica growing under high salt conditions (0.5 M NaCl) with salt free conditions as a control. Approximately 30â€40 million 76bp paired-end reads per sample were obtained using an Illumina NextSeq500. RNA-seq analysis was performed using Bowtie/TopHat/Cufflinks software pipeline. Total 15410 unigenes were generated with n50 value of 3038. A total of 13461 differentially expressed genes (fold change ≧ 2) were identified and 2646 genes were downregulated while 2446 genes were upregulated under high salt condition. We found that the genes involved in different cellular processes,such as metabolism, energy and biosynthetic processes, DNA repair, regulation of protein turnover, transport and saltstress tolerance were changed under high salt condition.
Conclusions: RNA-seq and pathway analyses found that salt stressed P. indica have significant differences in gene expression. Our results showed the complex mechanism of P. indica adaption to salt stress and it was a systematic work for endophyte to cope with the high salinity environmental problems. Thus, these results could be helpful for further investigation of the salt resistance mechanism in microbes.
Inkyu Park
Korea Institute of Oriental Medicine, South Korea
Title: Comparative analysis of complete chloroplast genome sequence of two Aconitum species and in the family Ranunculaceae
Biography:
Inkyu Park is a senior research scientist at the Korea Institute of Oriental Medicine (KIOM). He has completed his PhD from Chungnam National University at Republic of Korea. His research has centerd upon chloroplast genome study with development molecular marker and plastid evolution.
Abstract:
Aconitum species are well known herbaceous medicinal ingredient as well as toxic material and has great economic value in Asian countries. However, genomic information is still limited in Ranunculaceae. In this study, we completed chloroplast genome sequence of two Aconitum species, A. coreanum and A. carmichaelii, based on the Illumina miseq platform. The gene order, gene content and orientation of two Aconitum cp genomes exhibit the general structure of flowering plants, and are similar to other Aconitum species. The two Aconitum chloroplast genomes are 155,880 and 157,040 bp in length, respectively, and contain 131 unique functional genes including 86 protein coding gene, 8 rRNA and 37 tRNA. We established genetic relationship of Aconitum species and Ranunculaceae through phylogenetic tree based on 71 protein condign genes of 19 angiosperms. Comparison of the cp genome structure and gene order to those of Aconitum species revealed general contraction and expansion of the inverted repeat region (IR) and single copy boundary regions. We obtained barcoding target sequence and developed SCAR marker helpful for discrimination of the Aconitum species. These results suggest that the sequence variables of chloroplast genome could provide the useful genetic information and development of molecular marker for discrimination to identify Aconitum species
Zanenhlanhla Gumbi
University of KwaZulu Natal, South Africa
Title: Optimization of biomass and lipid production from a local Chlorella isolates using response surface methodology and artificial neural network
Biography:
Zanenhlanhla has completed a Bachelor of Science Honours degree, Cum Laude at the University of Kwa-Zulu Natal and is currenlty pursuing an MSc at the same insitution. She is a member of the Golden Key Honours Society and her key research interests lie in the fields of microalgal biotechnology and renewable energy.
Abstract:
The exhaustion of the world’s fossil fuel supplies and global warming are driving the search for renewable sources of fuel. Microalgae have received great interest as an alternative to fossil fuels due to their fast growth rates and high photosynthetic efficiencies. This study focuses on the optimization of biomass and lipid yield from an indigenous Chlorella isolate using the Response Surface Method. The input parameters consisted of NaNO3, NaHCO3 and NaCl within the ranges of 0.05-2.0g/l, 0.5-3.0g/l and 0-10mM respectively. Data from seventeen experiments with varied culture conditions was used to develop a polynomial model. Analysis of variance (ANOVA) of the model gave a coefficient of determination (R2) of 0.72. The predicted optimum conditions for biomass formation were 1.55 g/l NaNO3, 3.0 NaHCO3 and 0mM NaCl. The response graphs showing the interaction of NaHCO3 and NaNO3 on algal growth revealed that an increase in NaNO3 and NaHCO3 medium concentration enhanced the biomass formation whereas NaCl did not impact on biomass formation. These findings revealed that under optimal conditions the indigenous Chlorella isolate could be a potential strain for high biomass formation required for biodiesel production.
Preshanthan Moodley
University of KwaZulu Natal, South Africa
Title: A combined salt and acid pretreatment for enhanced enzymatic saccharification of waste sugarcane leaves
Biography:
Preshanthan Moodley has completed his Masters degree cum laude at the age of 27 from the University of KwaZulu-Natal in South Africa. His masters researched entailed exploring acidic pretreatment of waste sugarcane leaves for biohydrogen production by dark fermentation. He is currently studying towards his PhD degree with his research focusing on enhancing enzymatic sacharification of lignocellulosic waste towards bioethanol and biodiesel production.
Abstract:
Zinc cholride and sulfuric acid were employed as chemical catalysts to enhance enzymatic pretreatment of waste suharcane leaves.The effects of salt and acid concentration on the enzymatic digestibility using Novozymes Cellic Ctec 2 were examined at a lab scale. Leaves were pretreated using a combination of 3M ZnCl2 and 1.55 % H2SO4 (v/v) with a solid loading of 10% (w/v) at 121oC for 60 min. After washing, enzymatic saccharification was conducted with an enzyme and solid loading of 10 FPU and 10% respectively. Preliminary results indicated a glucose yield of 9.5 g/L per gram of dry weight sugarcane leaves. This yield showed an improvement over salt treatment and water treatment by 22% and 98% respectively. The next stage in this work will be to optimize the chemical (salt concentration, acid concentration and solid loadind) and enzymatic (enzyme loading, solid loading) pretreatment conditions. These findings illustrate the potential of low-cost chemical pretreatment to enhance glucose recovery from ligncocellulosic materials such as sugarcane leaves.
Yeshona Sewsynker
University of KwaZulu Natal, South Africa
Title: A two-stage hybrid pretreatment for enhanced enzymatic digestibility from corn cobs
Biography:
Yeshona Sewsynker has completed her MSc with cum laude at the age of 24 years from the University of KwaZulu-Natal. She is currently pursuing her PhD at the University of KwaZulu-Natal. She has published two of her Masters thesis chapters in peer-reviewed journals with the remaining two chapters currently under review.
Abstract:
This study focused on the effect of a combination of sulfuric acid and zinc chloride on the pretreatment of Corn cobs for sugar recovery and enzymatic digestibility. The first stage was a combination of zinc chloride and sulfuric acid which was autoclaved at 121ºC for 60 min. A solid to liquid ratio of 10% was used. The second stage was enzymatic hydrolysis using Cellic Ctec 2. Preliminary assessment of this hybrid pretreatment technique under a sulfuric acid concentration of 1.5%, zinc chloride concentration of 3M, enzyme loading of 10 FPU, and reaction time of 48 hr, resulted in a 75% increase in the glucose recovery compared to a single stage enzymatic hydrolysis. In addition, the two-stage method led to a 100% and 81% increase in the glucose recovery compared to the single stage zinc chloride and sulfuric acid pretreatments, respectively. These results evidently support that the combined ZnCl-H2SO4 with enzymatic pretreatment is an effective and feasible method for processing lignocellulosic biomass.
- Industrial and Microbial Biotechnology, Biotechnology and its Applications, Nano Biotechnology
Location: Sausalito
Session Introduction
Vinod Kumar
The University of Nottingham, UK
Title: Production of 1,3-propanediol from glycerol by mutant Klebsiella pneumoniae J2B devoid of 2,3-butanediol formation
Biography:
Currently, I am working as Marie Curie Fellow at Synthetic Biology Research Centre, The University of Nottingham, UK. I am working in the area of Biorefinery using Metabolic Engineering and Synthetic Biology tools for the sustainable production of biofuels and biochemicals through second generation biorefinery. So far I have published 19 research articles, two book chapters and two review articles. I have done my PhD (Biochemical Engineering & Biotechnology) and M.Sc (Chemistry) from Indian Institute of Technology Delhi, India. I have more than 13 years of research experience including my PhD and 5 year Post-Doctoral experience (France, S Korea, UK). In last 13 years, I got exposed to various areas of Biotechnology; Biochemical Engineering, Microbial Fermentations, Plant cell Technology, Agricultural Biotechnology, Metabolic Engineering and Industrial Biotechnology. I worked on different biological systems; fungal, yeast and bacterial and carried out research in multidimensional projects aiming at “Development of low cost, energy efficient and sustainable Bioprocesses for production of Biofertilizers, Biopesticides, Biofuels and Biochemicals”.
Abstract:
The 2,3-butanediol (BDO) is produced as a major byproduct during the production of 1,3- propanediol (PDO) from glycerol under limited aeration conditions by Klebsiella pneumoniae. In thepresent study, The BDO pathway genes, budA, budB, budC and budO (whole-bud operon), were deleted from K. pneumoniae J2B ΔldhA and the mutants were studied for glycerol metabolism and alcohols (PDO, BDO) production. Only the budO deletion mutant could completely abolish BDO production, but it exhibited serious reduction in cell growth and PDO production. By modifying culture medium such as increasing buffering capacity (from 29 mM phosphate to 100 mM phosphate) and adding bicarbonate (50 mM), the performance of the budO deletion mutant could be recovered to a similar level of the base strain (91.1 mM PDO under microaerobic condition) on flask scale. However, in fed-batch bioreactor experiment, the budO deletion mutant produced significantly less PDO (502 mM) than the base strain (753 mM). In addition, the budO deletion mutant produced significant amount of pyruvate (>73 mM) and lactate (>38 mM). The low PDO production in K. pneumoniae J2B ΔldhAΔbudO was attributed to the accumulation of glycolytic intermediates such as dihydroxyacetone and glyceraldehyde-3-phosphate, which are highly inhibitory to glycerol dehydratase.
Yogita Patil-Sen
University of Central Lancashire, UK
Title: Magnetoliposomes for hyperthermia cancer therapy
Biography:
Yogita obtained her PhD from the University of Manchester, UK and gained some postdoctoral experience at the University of Manchester and the University of Central Lancashire (UCLan). Currently, she is a Daphne Jackson Fellow at UCLan and her research is jointly funded by the Royal Society of Chemistry and UCLan. Yogita’s research interests are in the field of synthesis of different types of nanoparticles for targeted drug deliveryand cancer therapy. She has published nine research articles in high impact factor international journals. Yogita is a member of the Royal Society of Chemistry and the American Chemical Society.
Abstract:
Magnetoliposomes, hybrid nanoparticles made of superparamagnetic iron oxide nanoparticles (SPIONs)coated with liposomes, are emerging as new class of bio-nanomaterials due to their potential applications in targeted drug deliveryand hyperthermia cancer therapy. Coating SPIONs with liposomes enhances their biocompatibility anddispersibility and therefore their applicability in biomedical applications.The hyperthermia treatment is based on the fact that SPIONs, when subjected to an oscillating magnetic field generate heat and thus can kill tumor cells which are more sensitive to temperature above 41 °C than the normal cells. The amount of heat generated by SPIONs is strongly dependent upon their magnetic properties, which in turn are deteremined by their physico-chemical properties i.e. size and shape as well as coating material.
Herein we report the heating ability of bare SPIONs and core-shell type magnetoliposomes,which was measured using magnetic hyperthermia kit. SPIONs were coated with mixed lipid systems of phospholipids and cholesterol and the anticancer drug doxorubicin was encapsulated in the core-shell structure. The drug loading and release efficiency of bare and lipid coated SPIONs was also investigated. The results suggest that the drug loading efficiency increased upon lipid coating and drug release is much more controlled under the alternating magnetic field which indicates that magnetoliposomesare promising drug delivery vehicles for magnetic hyperthermia based cancer therapy.
Varsha Mehra
Shaheed Rajguru College of Applied Sciences for Women, India
Title: Synergistic antibacterial potential and total bioactive component determination of Elletaria cardamomum, Piper nigrum and Syzygium aromaticum
Biography:
Varsha Mehra has completed her PhD at the age of 28 years from University of Delhi . She is currently Assistant Professor in the Department of Biomedical Science, Shaheed Rajguru College of Applied Sciences for Women, a premier science college in the University of Delhi .She has a few publications in reputed international journals and currently engaged in Research Project on finding efficacious plant based drugs against Mycobacterium tuberculosis.
Abstract:
Spices are considered as rich source of bio-active antimicrobial compounds and are indispensable components of cuisines worldwide. They have been used since long to enhance the flavor and aroma of our foods. Besides, they also produce several medicinal effects and are used in treating various clinical ailments. To provide a scientific basis to traditional uses of Elletaria cardamomum, Syzygium aromaticum and Piper nigrum, their seed extracts as well as isolated phyto-constituents and combinations were evaluated for their antibacterial and antioxidant potential. Total phenol, flavonoid, condensed tannins and saponin contents were also measured. Organic extracts of all three spices showed good antibacterial activity against all the test strains, which was found to be comparable with standard antibiotics. Minimum inhibitory concentration for aqueous and organic seed extracts ranged from 25 to >50 mg/ml and 2 to 50 mg/ml respectively. Among the different extracts evaluated for DPPH free radical scavenging, ethanolic extract of S. aromaticum exhibited the highest inhibition with the IC50 value of 42 ± 7.4 µg/ml. This high radical scavenging activity can be directly correlated with the presence of high total phenolic content (310 ± 6.87 mg GAEs/g extract) possessed by the extract. Inhibitory activity of all the extracts was found to be increased, when used in combination. These findings suggest that these spices enhanced the functionality of the food in which they are used by effectively influencing their antioxidant and antibacterial potential.
Shahram Teimourian
Iran University of Medical Sciences, Iran
Title: Thwarting PTEN expression by siRNA, augments HL-60 cell differentiation to neutrophillike cells by DMSO and ATRA and reduces NETosis
Biography:
Shaharm Teimourian has completed his PhD at the age of 32 years from Tehran University and postdoctoral studies from Oxford University School of Medicine. He is the Director of medical genetics and biotechnology department. He has published more than 25 papers in reputed journals and has been serving as an Editorial board member of Edorium journal of molecular biology and world journal of hematology.
Abstract:
Abnormal cell differentiation,in particular suppression of terminal cell differentiation, exits in all tumors. Therapeutic interventions to restore terminal differentiation (“differentiation therapy”) are a very attractive way to treat cancer, especially leukemia.A variety of chemicals stimulates differentiation of leukemic cells, such as dimethyl sulfoxide (DMSO) and all-trans retinoic acid (ATRA). Tumor suppressor geneshave a vital role in thegateway to terminal cell differentiation.
In this study we inhibited PTENtumor suppressor gene expression by siRNA to investigate the effect of potentiating cell survival and inhibiting apoptosis on HL-60 cell differentiation by DMSO and ATRA at the same time we looked at Netosis.
Our results show that PTEN siRNA increasesHL-60 cell differentiation in the presence of DMSO and ATRA. At the same time the presence of siRNA hampered accumulation of apoptotic cells during incubation. PTEN siRNA reduced Net formation by differentiated neutrophils
Conclusion: Our study suggests potential usage of differentiation therapy in PTEN mutated AML leukemia.
Ryosuke Yamada
Osaka Prefecture University, Japan
Title: Acceleration of glycolysis and D-lactate production by novel global metabolic engineering in yeast
Biography:
R. Yamada has completed his PhD at the age of 27 years from Kobe university and postdoctoral studies from Kobe university. Then he moved as assistant professor to Osaka prefecture university. He has published more than 35 papers in journals related to applied microbilogy and biochemical enginnering.
Abstract:
The use of renewable feedstocks for producing biofuels and bio-based chemicals by engineering metabolic pathways of yeast Saccharomyces cerevisiae has recently become an attractive option. Many researchers attempted to accelerate glycolysis by overexpressing some glycolytic enzymes because most target bio-based chemicals are derived through glycolysis. However these attempts have met with little success. In this study, to create a S. cerevisiae strain with high glycolytic flux, we used multi-copy integration to develop a novel global metabolic engineering strategy. Then a novel global metabolic engineering strategy was applied for D-lactate production.
Among approximately 350 metabolically engineered strains, YPH499/dPdA3-34 exhibited the highest glucose consumption rate. This strain showed 1.3-fold higher cell growth rate and glucose consumption rate than the control strain YPH499/dPdAW. Real-time PCR analysis revealed that transcription levels of glycolysis-related genes such as HXK2, PFK1, PFK2, PYK2, PGI1, and PGK1 in YPH499/dPdA3-34 were increased. Besides, by using global metabolic engineering strategy, D-lactate was efficinetly produced.
This study successfully developed a novel global metabolic engineering strategy for S. cerevisiae, improving glucose consumption rate through optimizing the expression of glycolysis-related enzymes. The method detailed here is a promising approach to optimize S. cerevisiae metabolic pathways, thereby improving bio-based chemicals production using this organism.
Kamna Srivastava
University of Delhi, India
Title: Renin-angiotensin-aldosterone system genomics in essential hypertension
Biography:
Dr. Kamna Srivastava did her B. Pharm, M.Pharm and Ph.D. from Department of Pharmacology, Institute of Technology, Banaras Hindu University, INDIA. She held her Post-Doctoral positions in National Institute of Immunology; All India Institute of Medical Sciences, New Delhi. Presently, Dr. Kamna is Assistant Professor working in Molecular Cardiology Lab in Dr. B R Ambedkar Centre for Biomedical Research, Delhi University. Her on-going project is focused on the identifying the potential biomarkers for cardiovascular diseases. Dr. Kamna has more than 30 research publications to her credit and recipient of grants from DST, CSIR and ICMR India.
Abstract:
Objectives: Hypertension is one of the major cardiovascular diseases. Candidate genes encoding the Renin-angiotensin-aldosterone system (RAAS), ie, Angiotensin Converting enzyme (ACE), Angiotensinogen, Angiotensin II Type I receptor, Atrial natriuretic peptide (ANP) and Aldosterone synthase (CYP11B2), their expression at genetic and protein levels and their association with essential hypertension, if any, were investigated in a Northern Indian population.
Methods: Genotyping and Gene expression at mRNA and protein levels was carried out by PCR-RFLP, Real time PCR and Western blot respectively.
Results: A significant association was found in the AT1R genotypes (AC+CC) with essential hypertension. The expression of angiotensinogen was also upregulated in patients as compared to controls. The decreased levels of ANP gene expression at mRNA (85%) and protein (72.6%) levels and increased in CYP11B2 protein expression (1.53 fold) in the patient group as compared to controls were found. The individuals with rare allele in Angiotensinogen gene were found to have significant control in blood pressure with ACE inhibitor, Enalapril.
Conclusion: Our findings suggest the association of candidate gene of RAAS with essential hypertension. The increased expression of Angiotensinogen converting enzyme, Angiotensinogen, Angiotensin II Type I receptor gene and decreased levels of ANP gene expression at mRNA and protein levels in the patient group as compared to controls were significantly associated with essential hypertension and could be served as a prognostic biomarker for essential hypertension.
Hui Wang
Southeast University School of Energy and Environment, China
Title: New process for copper migration from contaminated soil and bioelectricity generation in soil microbial fuel cells
Biography:
Hui Wang has been studying for his PhD from Southeast University School of Energy and Environment. He majored in soil science and heavy mental pollution renovation. He has published one paper in reputed journals.
Abstract:
The soil microbial fuel cell (MFC) is a promising biotechnology for the bioelectricity recovery as well as the remediation soil. Moreover there were no studies on the heavy mental pollution in a soil MFC yet. A soil MFC was constructed to remediate the contamination soil and the electric field was generated from the oxidation of the acetate at the anode. We demonstrated that the copper migration, the power generation and the pH variation in the soil and the electrodes. The maximal voltage and the power density of 539 mV and 65.77m W/m2 were obtained in the soil MFC. The chemical fractionation of copper (Cu) was analyzed with a modified BCR sequential extraction method. The soluble Cu form and the total Cu contents from the anode to the cathode increased, and the difference between them kept growing over time. The Cu fractions in the soil and the electrodes were converted with the change of the dramatic pH from the anode to the cathode. There was a focusing effect leading to the change of the copper forms and the extractable acid form content increased in the three-fifths where the acid and the alkali fronts met.
Xian Cao
Southeast University School of Energy and Environment, China
Title: Simultaneous degradation of refractory organic pesticide and bioelectricity generation in a soil microbial fuel cell with different conditions
Biography:
Xian Cao study as a PhD candidate at School of Energy and Environment, Southeast University. His majored in microbial fuel cells (MFCs). He has published one paper in reputed journals.
Abstract:
In this study, the soil microbial fuel cells (MFCs) were constructed based on sandy soil to remove the refractory organic pesticide hexachlorobenzene (HCB) in topsoil by a simple method. The construction of membrane less single-chamber soil MFCs by setting up the cathode and the anode activated carbon, inoculating the sludge and adding the co-substrates can promote HCB removal significantly. The results showed that HCB removal efficiencies in the soils contaminated with 40 mg/kg, 80 mg/kg, and 200 mg/kg were 71.14%, 62.15% and 50.06%, respectively, which were 18.65%, 18.46% and 19.17% higher than in the control, respectively. The electricity generation of soil MFCs in different HCB concentrations were analyzed. The highest power density reached 70.8 mW/m2, and an internal resistance of approximately 960Ω was obtained when an external resistance loading of 1000Ω was connected. Meanwhile, the influences of temperature, substrate species and substrate concentrations on soil MFCs initial electricity production were investigated. The temperature between 25oC and 30oC had no influence on the initial electricity production in the soil MFC while the impacts of the substrate concentration were significant. The addition of the anionic surfactant sodium dodecyl sulphate (SDS) into the soil MFCs system contributed to the improvement of HCB removal efficiency.
Arezou Pakfar
Islamic Azad University, Iran
Title: Chondrogenic and possible pathologic effects of PRP on adipose derived MSCs
Biography:
Arezou Pakfar holds a Master degree in Cellular and Molecular Biology at Islamic Azad University, Iran. Her thesis was about Tissue Engineering and stem cells. She is working as a Researcher at Stem Cell Technology Research Center since 2014.
Abstract:
Introduction: Application of activated Platelet-Rich Plasma (PRP) with its vast range of cytokines and growth factors has achieved a considerable attention for chondrogenic differentiation in tissue engineering fields. Therefore, the aim of this study was to investigate the effects of PRP on human adipose derived MSC chondrogenesis.
Material & Methods: MSCs were differentiated using different PRP concentrations (5% and 15%). Changes in gene expression levels for cartilage and bone specific markers (COLII, AGC, SMAD2, SOX9) and (RUNX, Osteocalcin), respectively, were appraised by real time PCR. Also chondrogenesis was assessed by measuring secreted glucosaminoglycan in the medium or that kept in cell ECM. The expression of pathologic markers was evaluated by measuring the VEGF, TNFα secretion and alkaline phosphatase activity and calcium deposition.
Results: The most secreted VEGF (p<0.05) in 5% and 15% concentration were anti-angiogenesis. The inflammation factor (TNF-α) quantity of 5% PRP was the lowest (p<0.05) on 21st day but chemotaxic characteristics of the mentioned group was the highest. The expression levels of AGC, SOX9, COLII and RUNX were significantly (p<0.05) down-regulated while Osteocalcin was up-regulated. In addition, hypertrophy was seen in chondrogenic differentiation.
Conclusion: Due to having vast range of biologic active factors, PRP based chondrogenesis of human adipose derived MSC is dose dependent and the undesired outcomes due to absence of regulatory factors, should be suppressed by further optimizing the formulation of chondrogenic differentiation media.
- Poster Presentation
Location: Sausalito
Session Introduction
Yildiz Aka Kacar
Cukurova University, Turkey
Title: Determination of genes involved in lignification of pomegranate seeds by transcriptome sequencing
Biography:
Dr. Aka-Kacar has received her MSc. (1994) on plant tissue culture and PhD. (2001) degree on molecular characterization at Horticulture Department of Çukurova University in Adana,Turkey. During her PhD period she studied in Michigan State University, Plant and Soil Science Department, USA (TUBITAK-NATO Fellowship), East Malling Research Institute, UK, Bologna University, Horticulture Department Italy, as a visiting scientist. She spent a postdoctoral period at University of California Riverside, USA on genetic mapping in Citrus (2004-2005). She received Assoc.Prof title in 2006 and she has a full Prof position since 2013 at Cukurova University, Turkey.
Abstract:
Pomegranate (Punica granatum L.) is one of the oldest known edible fruit tree species, originating in Central Asia, but with a wide geographical global distribution. Besides using pomegranate as raw fruit, it has been used as herbal remedy. In consumption of pomegranate soft-hard seededness is very important. Soft seededness arises in a reduction of lignin. Lignin topo chemistry has ultra-structural aspects and lignification results from the enzyme mediated polymerization. Also lignin has three different monomers (coniferyl alcohol, sinapyl alcohol, p-coumaryl alcohol) are synthesized in the cytoplasm. Aim of the present study is to determine initiation time of lignification after pollination and genes involved in lignification mechanism in soft and hard seeded pomegranates. Known as a hard-seeded Hicaznar and soft-seeded 33N26 varieties were used as plant materials. Fruits from the two defined varieties were taken at intervals after pollination and fertilization at different sizes. Seed samples were used for transcriptome sequencing. Primary sequencing were produced by Illumina HiSeqTM 2000, called as raw reads, was subjected to quality control (QC). After QC, raw reads were filtered into clean reads aligned to the reference sequences. De novo analysis was performed to detect genes expressed in seeds of pomegranate varieties. We performed downstream analysis including gene expression, deep analysis based on gene expression, deep analysis based on DEGs, including Gene Ontology (GO) enrichment analysis. This dataset provides valuable information regarding pomegranate transcriptome changes for mechanism of soft-hard seeded pomegranate and may help guide future identification and functional analysis of genes that are important for lignification.
Wook Jin Kim
Korea Institute of Oriental Medicine, South Korea
Title: PNA analysis for authentication of four medicinal Paeonia species based on rDNA-ITS sequence
Biography:
Wook Jin Kim is currently a Senior Research Scientist at the Korea Institute of Oriental Medicine (KIOM). He works a development of DNA marker for discriminating betweenauthentic medicinal plant species and adulterants since 2012.
Abstract:
GennusPaeonia is an important medicinal plant in Asian traditional medicines. Among Paeonia species, P. lactiflora, P. japonica, P. veitchii, and P. suffruiticosa are pharmaceutically defined in different ways in the national pharmacopieias in Korea, Japan, and China.The roots of three Paeonia species, P. lactiflora, P. japonica andP. veitchii, commonly has been used as Paeoniae Radix, and the root bark of P. suffruticosahas been used as MoutanRadicis Cortex in Korean Traditional Herbal Medicine. However, only the roots of P. lactifolora and root bark of P. suffruticosa is pharmaceutically described as Paeoniae Radix and MoutanRadicis Cortex, respectively, in the pharmacopoeia of China and Japan.Since the morphological similarities of root and aerial part of these species, the identification of accurate species is very difficult. In addition, these herbal medicines bistributed as dried root slices or processed medicinal ingredients in the herbal market. Therefore, it is important to authenticate the different species used in these herbal medicines. So, we analyzed DNA barcode seqeunce of rDNA-ITS regionusing 17 samples of four Paeoniaspecies, and thenobtained species-specific marker nucleotides that can be used as genetic markers to identify these four plants at the species levels.Based on rDNA-ITS sequences, peptide nucleic acids (PNA) analysis which of probe-based fluorescence melting curve analysis was carried out to develop a powerful technique for detecting species-specific point mutations, namely marker nucleotide or single nucleotide polymorphism, capable to discriminate the fourPaeonia species without sequence analysis. Also, this method can provide rapid and efficient authentication of four Paeoniaspecies. Therefore, PNA analysis of four herbaciousPaeonia species will help to accurately authenticate each species and standardize the origin and quality of Paeoniae Radix and MoutanRadicis Cortex.
Birhanu Mekuaninte Kinfu
University of Hamburg, Germany
Title: Biocatalytic asymmetric phosphorylation catalyzed by recombinant glycerate-2-kinase
Biography:
Birhanu KINFU studied his BSc at the University of Gondar in Applied Biology and his MSc at Addis Ababa University in Biotechnology. He completed both degrees with great distinction. He has worked as university lecturer for 2 years. Winning the prestigious DAAD research grant award (German academic exchange service) under its ‘young academics and scientists’ program, he is currently doing his PhD at the University of Hamburg, Germany, Microbiology and Biotechnology department. His main work focuses on biocatalytic phosphorylation of selected metabolites, metagenomics and in vitro protein expression systems with manuscripts on pipeline.
Abstract:
D-Glycerate-2-phosphate is an important substrate and crucial metabolite of central carbon metabolism, glycolysis/ gluconeogenesis and pentose phosphate pathway, glycine, serine and threonine metabolism, methane metabolism, biosynthesis of plant secondary metabolites, phenylpropanoids, terpenoids and steroids, alkaloids derived from shikimate pathway, antibiotics, amino acids. Therefore an efficient, robust and scalable route for the preparation of enantio pure D-glycerate-2-phosphate is needed. A straightforward one-step biocatalytic phosphorylation of glyceric acid catalyzed by a recombinant glycerate 2-kinase heterologously expressed as maltose binding protein fusion has been investigated using racemic and the enantiopure D- and L-glycerate as substrate. The reaction was coupled with the phosphoenolpyruvate/ pyruvate-kinase-system for ATP-regeneration and monitored by 31P-NMR spectroscopy. This phosphorylation reaction using recombinant glycerate 2-kinase is highly enantio-selective and sustainable, as it yields enantiomerically pure D-Glycerate-2-phosphate in less reaction steps and with higher purity than chemical routes.
Wojciech Hahnel
Hochschule Kaiserslautern University of Applied Sciences, Germany
Title: Development of an automated portable system for extraction of DNA from difficult samples
Biography:
Wojciech Hahnel, 28 years old, has completed his B.Sc. in Biotechnology in 2011 at Mannheim University of Applied Sciences, Germany, followed by a M.Sc. in Biotechnology with focus on “Bioprocess Development” in 2013 at the same university. Since 2014, Wojciech Hahnel is doing his PhD at University of Applied Sciences Kaiserslautern, Germany, which he is planning to finish in 2017. During his studies he has been working at the Dublin City University (DCU) in Dublin, Ireland, and at the University Hospital in Bochum, Germany.
Abstract:
DNA extraction from difficult samples (e.g. stool, soil) is problematic, as there are inhibitors that influence further processing of DNA and compounds which are mutagenic and destructive against DNA. Especially fecal DNA has diagnostic relevance because it contains DNA from various sources, like blood or intestinal mucosa. Analysis of fecal DNA might allow conclusions on the presence of (intestinal) diseases like tumors and inflammations at early stages in a quick and non-invasive manner.
Kits for DNA extraction from difficult samples are commercially available, but these are neither automated nor quick nor easy to use. Thus, an easy automated portable system for extraction of DNA from difficult samples would be beneficial. It could be used on-site, e.g. bedside in hospitals.
After testing different strategies and kits, the geneMAG-RNA/DNA kit from chemicell using magnetic beads was found to be suitable. Since this kit has been developed for other sample types, it was adapted and scaled down to a portable microchip system. For further evaluations soil was used because of easier availability and handling. Centrifugation was replaced by sedimentation. As a result complete DNA extraction could be performed inside the chip. As pumping is performed by air pressure, most of the tubing is not in contact with liquids and can be reused. The chip itself can be reused as well.
Future plans include further automation of the system, testing of different sample types and evaluation of the sedimentation step. Secondly, we test pre-filled chips in order to provide a ready-to-use system.
Araceli Olivares
Pontifical Catholic University of Valparaiso, Chile
Title: Effect of simulated gastric juice over viability in microencapsulated Lactobacillus casei
Biography:
Araceli Olivares is a Biochemical Engineer at the Pontifical Catholic University of Valparaiso (PUCV), Dr. in Engeniering Science m/Biochemical Engineering in PUCV. Since 20011 is a researcher of the Center for Studies on Healthy Food (CREAS). She has 3 published patents (one in Chilean and two in Unites State Patent Ofice) and 5 application patents in USPTO. She has 8 scientific journals.
Abstract:
Probiotics must be delivered alive to exert a positive health effects in site of action (the intestine). Once reach the intestine, they should establish themselves and exert a positive health effects. They must be survive the stomach acidic, bile acid and others degradative enzime througth intestinal tract.
The aim of current study was to evaluate the performace of microencapsulated Lactobacillus casei using alginate gel matrix to facilitate a suitable carrier system to enhance this objetive and confer protection from the acidity.
A solution with lyophilized L. casei at 5g/L (108 CFU/mL) was prepared and mixed (1:1) with sodium alginate solution at 2%. As a hardening solution, calcium chloride at 0.1M with gently stirring was used. Microencapsulator provide by BÜCHI (Encapsulated B-390) was used.
Simulated gastric juice (SGJ) with 9g/L of sodium chloride and 3g/L of pepsin was prepared. Different pH was adjusted with 1M HCl. 400mg of microspheres containing L. casei were mixed in 20mL of SGJ with pH adjusted at 2.0; 2.5; 3.0; 3.5 and 4.0 and incubated until 120 min a 37oC and 50rpm. Microencapsulated Lactobacillus were filtered and dissolved in sodium citrate to released and count CFU in supernatant.
The best results were obtained at pH 4.0 (over 70% of survival at 90min). At pH 3.5 the survival was over 20% at 60 min, however at pH less than 3.0 there was a rapid loss of viability and the survival was null at 90 min. Then, a microencapsulated is an available technique to maintain viability of probiotics to protect the passage through intestinal tract.
Acknowledgment: Project Fondecyt No 11130460
Jee-Yeon Ryu
Ajou University, South Korea
Title: Nanoliposome particle possessing protein based CRISPR/Cas9 system for the therapeutic applications
Biography:
Jee-Yeon Ryu is a PhD candidate from Ajou University College of Pharmacy. She is the director of Tae-Jong Yoon. She is studying synthesis of nanomateials aimed at biological applications..
Eun Yi Cho has completed her PhD from Chonnam National University, South Korea in 2011 where she majored biochemistry and then she is post-doctoral researcher at the Ajou University College of Pharmacy through the korea research institute of chemical technolgy (KRICT). She is currently the study of gene edting using nanotechnology.
Abstract:
Gene editing (CRISPR/Cas9) technology has been spotlighted as a superior therapeutic tool that is capable of treating the fundamental causes of disease induced by genetic abnormalities, which have to date been considered as incurable diseases. Thus many researchers have focusing establishment of gene editing moiety delivering, because efficient and safe delivery in the body remains one of the major challenges of biomedical and nano-pharmaceutical research. A plasmid based CRISPR/Cas9 system has shown several critical limitations such as off-targeting, integration of DNA segment and toxicity of transfection agent. To overcome these problem, protein based CRISPR/Cas9 system was recently co-opted to the therapeutic or gene editing method. However, the protein system has still remained the stability problems, especially in in vivo system as like degradation by enzymatic reaction or low efficiency.
Herein, we elucidate novel method for the delivery system of protein based CRISPR/Cas9 with high efficient and bio-compatibility. The CRISPR/Cas9 complex was successfully encapsulated into the nanometer sized liposome (nanoliposome), which was composited with bioapplicable phospholipid chemicals through metal coordiation reaction. After chemical surface modifcation, the nanoliposome with gene editing materials was shown long term solution stability without agglomeration and penetrated well into the cell cytosol. We suggest optimization of preparation for the nanoliposomal protein based CRISPR/Cas9 system as a platform particle to the therapeutic application. As a proof of concept, the nanoliposome with CRISPR/Cas9 system was exploited to apply for the type 2 diabetes therapy and exhibited effectiveness of glucose control without off-targeting, acute toxicity and degradation. More importantly, the our particle platform system was shown higher regulation effect than a clinical used chemical drug without various adverse reactions effect including renal disorder or allergic reaction.
Biography:
The presenting author, Gourab Ghosh is currently pursuing his Ph.D. in transgenic crop science from St. Xavier’s College, Kolkata affiliated under University of Calcutta, India. He is 27 years of age. He has two publications in reputed, peer-review journals to his name.
Abstract:
Pigeonpea is one of the major grain legumes of tropics and subtropics, covering vast regions of developing countries from Africa, Asia to Latin America. It ranks fifth in area among pulses after soybean, common bean, peanut, and chickpea.Globally, pigeonpea is cultivated on 4.92 million hectares with an annual production of 3.65 metric tons and productivity of 898 kg/ha2.As they are grown in harsh environments and exposed to a variety of biotic and abiotic stresses, their productivity has not increased conspicuously for the last 50 years.Among many insect pests, the pod borer Helicoverpa armigera causes significant damage to this crop. It is the most devastating Lepidopteran pest and causes extensive economic losses to the tune of US$ 300 million annually. The present study seeks to protect pigeonpea plants from H. armigera infestation by incorporating cry1Ac and cry2Aa genes, through a unique and efficient gene transfer method. An Agrobacterium tumefaciens-mediated transformation strategy was implemented using in vitro transgenic shoot-grafting technique. A. tumefaciens harbouring different binary vectors containing cry1Ac and cry2Aa geneswere used for transgenic pigeonpea development. An overall 7-9% of transformation frequency was recorded. After monitoring transgene integration by Southern hybridization, transgenic T1 and T2 lines were further analyzed by western blot, ELISA and insect bioassay. Transgenic lines obtained,exhibited optimum expression of Cry1Ac and Cry2Aa proteins. This study was further extended to the development of selectable marker (nptII) free cry1Ac expressing transgenic lines using cre-lox mediated marker elimination system
Miyu Nishiguchi
Nara Institute of Science and Technology, Japan
Title: Ergothioneine fermentative production in Escherichia coli
Biography:
Miyu Nishiguchi is second year master’s student in NARA INSTITUTE of SCIENCE and TECHNOLOGY. She study applied microbiology.
Abstract:
Ergothioneine (ERG) is sulfur-containing amino acid synthesized by certain bacteria and fungi. Recently, findings point to critical functions in human physiology. Human takes ERG from food and concentrates it in specific tissues or cells such as liver, kidney, central nervous system, and red blood cells. ERG is marketed as dietary supplement or nutraceutical so that acts as anti-oxidant. It has been recognized that filamentous fungi or Actinomycetes produce ERG. However, in 2010, the ERG biosynthetic gene was identified for the first time. Here, we challenged to produce ERG from glucose with our constructed cysteine producer. E. coli has a regulation system that synthesized cysteine from energetically-favored thiosulfate, as the assimilation of sulfate spends 2 ATP and 4 NADPH. This cysteine producer produces 16 g/L of cysteine from thiosulfate. Therefore, we established world-first ERG fermentation and challenged production of more cheaper ERG.
We cloned ERG biosynthetic genes from Micobacteria smegmatis, and performed heterologous expression of cloning ERG genes in E. coli. The analysis of the culture medium by LC-MS/MS detected ERG peak. When a plasmid carrying these ERG biosynthetic genes was introduced into cysteine producer with enhanced biosynthesis, weakened degradation, and improved export of L-cysteine, the transformants slightly produced ERG into medium from thiosulfate (30mg/L of ERG). Interestingly, this transformants produced 200 mg/L of ERG from sulfate. We propose that the spend of NADPH is important for production of ERG.
(This work was supported by the Programme for Promotion of Basic and Applied Researches for Innovations in Bio-oriented Industry to I.O.)
Rayssa Almeida Garcia
Federal University of Brasilia, Brazil
Title: Strategy to overcome nucleic acid degrading enzymes in insect pests for the RNAi application
Biography:
Rayssa Almeida Garcia is a PhD student in the Molecular Biology Program of the Federal University of Brasilia – Brazil, at the age of 23. She does her research work at in the Plant-Pest Interaction laboratory at Embrapa Genetic Resources and Biotechnology- Brasilia/ Brazil, under supervising of Dr Maria Fatima Grossi-de-Sa, have published a large number of papers in reputed journals and is a fellow member of the World Academy of Science.
Abstract:
Gene silencing through RNA interference as a biotechnological approach for the control of crop insect-pests have been intensively applied in the last few years. dsRNA microinjection and in vitro feeding are the most wildly used approaches for administering RNAi in insects. However, RNAi efficiency appears to be variable among different insect groups, especially when applied by feeding - for some insect groups the oral delivery of the dsRNAs has been reported highly ineffective. In initial studies, our gene silencing data for cotton boll weevil (Anthonomus grandis) were unclear when dsRNA administration was done by feeding. The purpose of this work was to assess the possibilities of RNAi as a tool for the control of this insect pest using oral delivery of dsRNAs and to investigate the reason for the low efficiency in gene silencing, aiming to develop a strategy to deal with the efficiency and usage of dsRNA by feeding. Data showed an optimal nucleasic activity of the A. grandis gut nucleases at acid pH, ranging from 5.5 to 6.5, and the A. grandis' gut homogenate significative degraded both dsRNA and dsDNA. Three nuclease sequences were found in A. grandis transcriptome, named AgNuc1, AgNuc2, and AgNuc3; in which AgNuc2 and AgNuc3 showed to be highly expressed in the insect gut. The silencing of the three nuclease genes strongly diminished dsRNA degradation when dsRNA were incubated with homogenate from silenced insects. On the other hand, when dsRNAs were protected with a Cell Penetrating Peptide (CPP) fused with a dsRNA Binding Domain (DRBD), no dsRNA degradation was found. Furthermore, dsRNAs complexed with CPP-DRBD were found to enter into A. grandis gut cells. The dsRNA complexed administered in the diet caused a greater gene silencing, compared to naked dsRNA. All data point out to the relevance for overcoming the gut nucleases with/or in parallel with the RNAi applications for the control of crop insect-pests.
Eman Bellah Aly Ramadan
The American University in Cairo, Egypt
Title: Molecular adaptation of a metagenome-derived mercuric reductase from Kebrit Deep brine environment in the Red Sea to high salinity
Biography:
Eman Ramadan finished Bachelor of Medicine, Bachelor of Surgery (M.B., B.C) Ain shams university faculty of Medicine in 1997. Then finished M.Sc. Degree in pediatrics from Ain shams university faculty of Medicine 2002. In 2011, received M.Sc. in Biotechnology from American university in Cairo. Currently PhD Candidate: American university in Cairo Egypt, expected graduation fall 2016.
Abstract:
Red SeaKebrit Brine (1490m)possess unique environmental conditions, characterized by high salinity 4 M, temperature 23.4°C, elevated concentration of heavy metals, no oxygen and high hydrostatic pressure. In order to highlight the structural-functional relationship of enzymes adaptation to such extreme environmental conditions, DNA isolated from the microbial community of Kebrit Brine is subjected to 454-pyrosequencing and a metagenomic dataset is established; and looked for enzymes involved in mercury detoxifications. An operon containing the genes essential for mercury detoxification was identified in our 454-pyrosequencing metagenomic dataset. A total of 28merAOrthologs were identified in Kebrit Brine metagenomics library, choosing two merA genes: one representing the consensus sequence (K35-NH) and the other (K09-H) has amino acid substitutions replacing non-polar with acidic amino acids. Kinetic parameters were measured at the NaClconcentration that gave maximum activity for the respective enzyme. K09-Hmaximal activation is observed at 2Molar NaCLand retains 65% of its activity at 3 Molar. K35-NH showed maximum activity at 0 Molar NaCl which is equvlent to 52% activity of K09-H at 4 M. Both and K35-NH retained 90%of their activity after 10 minutes incubation at 65ËšC.Comparing the kinetics of both enzyme suggested, that the amino acids difference between the two orthologs are acquired evolutional structural adaptation to confer site-specific level of halophilicity to survive in such extreme environment as KebritDeep.Thishalophilic feature if used properly is a potential for many industrial and bioremediation applications in detoxification of mercury.
Shota Kajiwara
Osaka Prefecture University, Japan
Title: Development of lipase-sucrose complex to improve of transesterification activity and stability in the organic solvents
Biography:
Shota Kajiwara was recieved the Master of Engineering degrees in Osaka Prefecture University, Osaka, Japan, in 2016. He is now a doctoral course student of Osaka Prefecture University. His research interests includes the stabilization of enzyme in the presence of organic solvents.
Abstract:
Lipases are one of the most important enzymes as industrial catalysts. When lipases are used in the presence of organic solvents, the reaction rate is remarkably increased by solubilizing the poorly soluble substrates. Furthermore, lipases can catalyze ester synthetic reactions and transesterification reactions in the presence of organic solvents. However, enzymes including lipases are generally inactivated in the presence of organic solvents by direct contact with molecules of organic solvents. Some coordinated water molecules exist around sugars such as sucrose. Thus when lipase was complexed with sucrose and used in the presence of organic solvents, coordinated water molecules around the lipase-sucrose complex might reduce direct contact between lipase and organic solvent molecules. In this study, the transesterification activity of the lipase-sucrose complex in the presence of organic solvents was examined. The lipase-sucrose complex was prepared by freeze-dry of a lipase solution containing 1% (w/v) of sucrose (mass ratio of lipase:sucrose = 1:130). The lipase-sucrose complex showed the higher transesterification activity than freeze-dried lipase without sucrose in the presence of n-hexane and n-hexane containing 50% (v/v) of n-octane, 1-propanol, 1-pentanol, or 1-octanol. After incubation in 100% of n-hexane, n-octane, or 1-propanol at 30ËšC for 24 hours, the residual transesterification activity of lipase-sucrose complex was higher than that of the freeze-dried lipase without sucrose. Furthermore, after incubation in 100% of 1-pentanol or 1-octanol, the transesterification activity of lipase-sucrose complex was also increased. In conclusion, the transesterification activity and organic solvent-stability of the lipase were successfully improved by complexation with sucrose.
Mohamad Maged Galal
The American University in Cairo, Egypt
Title: Functional adaptation of mercuric reductases from the deep brine environment of Atlantis II in the Red Sea to high temperature
Biography:
MohamadMaged has graduated from the Faculty of pharmacy, Cairo University in 2007. He has completed his MSc.in Biotechnology at the American university in Cairo (AUC) in 2011 and expected to finish his PhD from the AUC in December 2016. He is also a nutritionist and a personal trainer.
Abstract:
The lower convective layer (LCL) of the Atlantis II brine pool of the red sea is a unique environment characterized by high salinity of 4.4 Molar, temperature of 68οC and very high concentrations of heavy metals. Mercuric reductase enzymes functional in such extreme conditions could be used inthe environmental detoxification of mercurial poisoning. This would be of use in the Egyptian Gold mines, where mercuryused in the extraction process imposes a serious ecological hazard.We constructed an environmental Atlantis II mercuric reductase library, where we have identified two classes of mercuric reductases. One is the halophilic/thermostablemerA. It is designated ATII-LCL-H. It retains 50% of its activity after 10 minutes incubation at 75ËšC. The other is a non-halophilic /thermostablemerAdesignated as ATII-LCL-NH. Itretains 61% of its activity at 65ËšC.The ATII-LCL-HmerAhas two characteristic signature boxes and a short motif composed of 4 Aspartic acids (4D414-417). In order to understand how two enzymes from the same environment have evolved to withstand heat, we mutated the isoform ATII-LCL-NH.Substitution of 2D at positions 415/416 enhanced the thermal stability while other mutants did the opposite effect. The 2D mutant retains 88.6% of its activity at 65ËšC. Three-dimensional structure prediction revealed newly formed salt-bridges and H-bonds in the 2D mutant as compared to the parent molecule.
Biography:
Abril Gomez-Guzman is currently a graduate student in the University of Guadalajara Jalisco, Mexico. She is doing her Ph.D studies in the program of Science of Biotechnological Processes. She has completed her master studies from the Autonomous University of Baja California at the age of 25 year and her bachelor degree studies from the Technological Institute of Tepic, in Biochemical Engineering. She was a professor of the Autonomous University of Guadalajara in the period of 2011-2013. She has published one paper in reputed journals and she is working in another ones.
Abstract:
Microalgae and some bacteria offer a promising technology for the removal of nutrients such as Nitrogen and Phosphorus in municipal wastewater. In this investigation the microalgae Chlorella vulgaris, the bacteria Pseudomonas putida, Bacillus cereus and the consortium microalga/bacteria of these microorganism were used as model in the removal of nutrients using a model wastewater. The results of removal of nutrientes with Chlorella vulgaris was 24%, 80.6% and 4.30% for NO3-N, NH3-N and PO4-P, respectivley. For Bacillus cereus the removal of nutrients was 8.4%, 28.8% and 3.8% of NO3-N, NH3-N and PO4-P. With Pseudomonas putida was 5%, 41.8% and 4.3% of NO3-N, NH3-N and PO4-P (in 168 hours respectively). The consortium of Chlorella vulgaris/Bacillus cereus/Pseudomonas putida were able to remove the 29.4%, 4.2% and 0% of NO3-N, NH3-N and PO4-P in 240 hours. The highest biomass production was found with Bacillus cereus 450 mg/L followed by Pseudomonas putida 444 mg/L, the consortium 205 mg/L and Chlorella vulgaris 88.9 mg/L. This study showed that Chlorella vulgaris consume first NH3-N as a source of Nitrogen and when it is exhausted then consume NO3-N, it happen when both compounds are available in the medium. In the removal of phosphate (PO4-P) the microorganisms Chlorella vulgaris and Bacillus cereus were able to remove a higher amount because they can assimilate this nutrient even in stress conditions (like changes in pH) such as happening in the municipal wastewater.