Day 2 :
Keynote Forum
Michi Umetani
University of Houston, USA
Keynote: Modulation of estrogen receptor activity by 27-hydroxycholesterol and its mechanisms of action
Time : 09:30-10:05
Biography:
Michi Umetani has completed his PhD at the University of Tokyo and postdoctoral studies from University of Texas Southwestern Medical Center. He is Assistant Professor of the Center for Nuclear Receptors and Cell Signaling, University of Houston. He has been working on the impact of cholesterol metabolites in cardiovascular and metabolic health and disease.
Abstract:
27-Hydroxycholesterol (27HC) is one of the most abundant oxysterols in human circulation, and its levels correlate with that of cholesterol. Previously, we found that 27HC directly binds to estrogen receptors (ER) and works as a selective estrogen receptor modulator (SERM). 27HC is the first identified, endogenous SERM, and in addition to blocking the cardiovascular protective effects by estrogen, it induces vascular inflammation and augments atherosclerosis independently of estrogen action. Thus, 27HC is an important factor involved in the atherosclerosis development in hypercholesterolemia. Using synthetic biological approach, we investigated the mechanism by which 27HC modulates ER activity. This project has the potential to develop a novel therapeutic intervention to prevent hypercholesterolemia-derived cardiovascular dysfunction.
Keynote Forum
Mehmet Sen
Harvard Medical School, United States
Keynote: Structural alterations of leukocyte integrin, LFA-1
Time : 10:05-10:40
Biography:
Mehmet Sen has completed his PhD from the University of Houston, Department of Biology and Biochemistry with Glen Legge, M.D., Ph.D., and postdoctoral studies from Harvard Medical School, Department of Biological Chemistry and Molecular Pharmacology with Timothy Alan Springer, Ph.D.. His research interests lie in the structural and functional basis of receptor/ligand interactions, which are relevant to human health and disease. Structural studies by protein NMR spectroscopy, X-ray Crystallography and Electron microscopy are complemented by functional approaches using molecular biology and protein engineering to dissect structural information, design proteins/peptides/small molecules with modified specificities and activities, and ultimately contributes to the development of biologics with therapeutic potential.
Abstract:
The mechanism through which adhesiveness of lymphocyte function-associated antigen-1 (LFA-1) is the consequence of highly coordinated structural rearrangements within both α and β subunits yet, atomic details of LFA-1 conformations remain to be investigated. Here we determined the crystal structures of the LFA-1 headpiece that shows coordinations of cations in the closed conformation. The external ligand binding (αI) domain provides striking interdomain flexibility, which potentially plays a pivotal role for greater accessibility in ligand recognition. Loops at the αI and β-propeller interface for the αL and αX differ in length, orientation and post-translational modifications. Moreover we also characterize in atomic details of how a shape-shifting pathway of the β2 I-domain regulates its affinity for the internal ligand, which occurs in a different order than in β1, β3 and β6 integrins. Small angle x-ray scattering (SAXS), negative stain electron microscopy (EM) and adhesion assays here show that LFA-1 activation requires headpiece opening event. Mn2+, universal integrin activator together with the high-affinity ICAM-1 ectodomain was examined for the effect on the equilibrium between the open and closed LFA-1 headpiece conformations. Mn2+, relative to the Mg2+ shifted the equilibrium toward the open conformation. Addition of ICAM-1 substantially stabilized the open headpiece. The observed headpiece opening upon addition of Mn2+ and ICAM-1 is consistent with our previous structural studies and headpiece-opening model.. Furthermore, our adhesion assays revealed that an activating and inhibiting βI mutations stabilized the open/extended and bent/closed integrin states, respectively, which provides a structural mechanism into a leukocyte adhesion deficiency (LAD-I) mutation.
- Workshop
Location: Canterbury
Session Introduction
Vsevolod V (Seva) Gurevich
Vanderbilt University, USA
Title: Mass production of high fidelity oligonucleotides as feed stock of Post-genome-sequencing synthetic biology (PGS-SB)
Time : 11:00-12:30
Biography:
Vsevolod V (Seva) Gurevich has completed his PhD in 1989 from Shemyakin Institute of Bioorganic Chemistry, Moscow, Russia, and postdoctoral training (1991-1995) in the laboratory of Dr. J L Benovic, Thomas Jefferson University, Philadelphia, PA. He is Professor of Pharmacology at Vanderbilt University. He has published more than 160 papers in reputed journals and has been serving as an editorial board member of several journals.
Abstract:
Virtually every protein is multi-functional. Over-expression or knockdown/knockout by definition enhance or suppress all functions of targeted proteins. Thus, these methods are inherently unsuitable to elucidate biological role of individual functions. Re-engineered of signaling proteins to enhance or suppress a single function out of many is a viable alternative. However, in contrast to over-expression or knockout, this approach requires in-depth knowledge of the molecular mechanisms of each function of the protein of interest. Relatively few proteins are studied in sufficient detail to make this possible. We will discuss this approach using arrestin proteins as an example. Arrestins are ultimate multifunctional scaffolds, organizing multi-protein signaling complexes and localizing them to particular subcellular compartments. Extensive mutagenesis and structural work revealed the molecular basis of at least some arrestin functions, enabling the construction of signaling-biased arrestins, as well as the use of their mono-functional elements to elucidate the biological role of their functions. Arrestins regulate G protein-coupled receptors, cell adhesion and motility, as well as MAP kinase signaling involved in life-or-death decisions of the cell. Therefore, reengineered “designer” arrestins and their parts are useful tools for experimental manipulation of cell behavior. Practically every disorder is associated with faulty signaling. Signaling-biased arrestins and other regulatory proteins allow us to channel cell signaling in desired direction, which has enormous therapeutic potential.
Weiwen Zhang
Tianjin University
Title: Synthetic photosynthetic Cyanobacteria to produce fine chemicals directly from CO2
Biography:
Dr. Zhang is a Professor of Microbiology and Biochemical Engineering, Head of Laboratory of Synthetic Microbiology in the School of Chemical Engineering and Technology of Tianjin University in China. Prior to joining Tianjin University, Prof. Zhang was a faculty with Biodesign Institute and the School of Life Science of Arizona State University for 2007-2011, and a senior scientist at Microbiology Department with the Pacific Northwest National Laboratory of the U.S. Department of Energy (DOE) for 2002-2007. Dr. Zhang has broad research experience in microbial genetics, physiology and synthetic biology, has authored more than 130 peer-reviewed scientific papers. His recent research focuses on synthetic biology of photosynthetic microorganisms and analysis of microbial consortia.
Abstract:
Photosynthetic Cyanobacteria have attracted significant attention as a “Microbial factory†to produce biofuels and various fine chemicals, mostly due to their abilities to utilize CO2 and sunlight directly as carbon and energy sources, respectively. We will report our recent progress on engineering cyanobacterium Synechocystis sp. PCC 6803 to produce 3-hydroxybutyrate (3HB), 3-hydroxypropionic acid (3HP) and butanol directly from CO2, and on strengthening Synechocystis for better tolerances against toxic products.
Mateus Schreiner Garcez Lopes
Innovation in Renewable Technologies - Braskem – Brazil
Title: Driving innovation with renewable chemicals platform - the role of synthetic biology in the petrochemical industry
Biography:
Mateus Schreiner Garcez Lopes has completed his PhD at the age of 27 years from University of São Paulo/Brazil and University of Erlangen/Germany. His postdoctoral studies were at University of São Paulo and he holds a MBA in Agribusiness also from the University of São Paulo. He had worked as researcher at the Natural Energy Institute (Hawaii- USA) and the Institute of Biotechnology from UNAM (Mexico). He is responsible for the Innovation in Renewable Technologies at Braskem, the largest petrochemical producer in the Americas and the world's leading biopolymer producer. He has published in international journals, has issued patents on new metabolic pathways and is serving as an industrial board member for Synberc (Synthetic Biology Research Center - USA).
Abstract:
Braskem reinforce the leadership in bio-thermoplastic, by signing a development agreenment with Novozymes in 2012 for the development of bio-polupropylene. Both world-class companies are working together for the reinforce Braskem vision in the thermoplastics segment. Going forward into other markets, Braskem design a bio rubber platform in partnership with two synthetic biology start-ups. The company have signed an agreement for the joint development of two new technology for the production of butadiene and isoprene from renewable feedstocks with Genomatica (signed in 2013) and Amyris (signed in 2014), respectively. Through this partnership, Braskem reaffirms its commitment to invest in the research of producing chemicals from renewable feedstocks, effectively strengthening its leadership in synthetic biology. Braskem has invested in one of the most advanced laboratory in South America. A 7000 ft2 facility with a m Braskem vision is to be world leader in sustainable chemistry, innovating to better serve people. To transform this vision into reality, we are believe that synthetic biology will empower new technologies and transform our society. I am current responsible for biotech innovation at Braskem and my lecture will present why we choose synthetic biology as a platform for industrial biotechnology and what is our strategy to move forward in this field. ultidisciplinary and international team in a variety of areas, including synthetic biology, protein engineering, automation, bioinformatics, fermentation and downstream processes. The research program conducted within Braskem seeks not only to find alternatives that are based on renewable feedstocks, but on developing routes that are also competitive in terms of production cost.
Teresa Cristina Zangirolami
Federal University of Goiás, Brazil
Title: Improved ethanol production from D-xylulose through evolutionary engineering of wild baker´s yeast
Biography:
Teresa Cristina Zangirolami has completed her PhD from Technical University of Denmark. She is a Lecturer and Researcher at the Chemical Engineering Department of Federal University of São Carlos and works in close cooperation with researchers from Butantan Institute and from the Department of Biological Engineering (University of Minho, Portugal). She has published more than 30 papers in reputed journals and has been serving as an Editorial Board Member of repute.
Abstract:
For enhanced economic feasibility of second generation bioethanol production, it is important to use all the fermentable fractions present in sugarcane bagasse (cellulose-C6 and hemicelluloses-C5) or other feedstocks. Xylose is the main sugar found in the C5 fraction and is not assimilated by Saccharomyces cerevisiae. However, this yeast is able to ferment D-xylulose, an isomer of xylose which can be obtained using glucose isomerase generating ethanol and xylitol as main products. This study aims to select a more suitable ethanol producer strain exhibiting lower xylitol production. Colonies isolated from industrial yeast were exposed to a sequence of evolutionary engineering procedures: Incubation on complex solid media containing xylulose (aerobic conditions); incubation of selected colonies in minimal solid medium (anaerobic conditions) and cultivation of 20 adapted colonies in cuvettes containing YNB medium and xylulose. Three colonies exhibiting lowest xylitol production were identified and characterized in terms of product formation and substrate uptake under micro aerobic conditions at 30° C. selected mutant and wild-type strains were further compared in terms of morphology and genotype. With a final ethanol concentration of 4.3 g/L for a xylitol production of 0.55 g/L, the adapted yeast showed selectivity values up to 26 mol ethanol/mol xylitol. The morphology analysis revealed that mutant strain was not able to metabolize bromocresol green dye and its colonies exhibited a darker green color on WLN agar. The results of genotypic analysis also confirmed that evolutionary engineering was able to introduce several mutations in the selected yeast.
Masoud Sheidai
Shahid Behesti University, Iran
Title: Olive biotechnology: The assessment of tissue culture induced genetic variability in olive (Olea europaea) by molecular markers
Biography:
MASOUD SHEIDAI has completed his PhD at the age of 28 years from Poona University India and Sabbatical studies from UBC University in Canada. He is now proffessor of Biology in Shahid Beheshti University, Tehran, Iran. He has published more than 125 papers in reputed journals and has been serving as an editorial board member of repute. Gene Conserve, and Bidiversity journals.
Abstract:
Olive is one of the most important tree crops of the country and it is cultivated in many places. Propagation of true to type olive plants is important for many olive companies and farmers in the country and for this reason genetic analysis of randomly selected tissue culture regenerated olive plants of cultivars Kroneiki, Zard, Roughani and X was performed and compared with the mother plant. We used Cp-DNA and RAPD molecular markers to check genetic fidelity versus somaclonal variation. Both RAPD and Cp-DNA analyses revealed that some of the regenerated plants differed extensively in their genetic content. These plants were placed in different clusters far from the mother plants in NJ tree and TNT tree obtained from these two molecular markers. We identified also some regenerated plants that were true to types of the mother plants. These plants also differed in their morphological features. Genetic and morphological changes were more extensive with increase in sub-culture numbers. Different uses of these two types of plants will be discussed.
- Molecular Modeling and Drug Designing | Development of Recombinant-DNA technology | Computational Biology | Animal Cell and Tissue Engineering
Chair
Vsevolod V (Seva) Gurevich
Vanderbilt University Medical Center, USA
Co-Chair
Mehmet Sen
Harvard Medical School, USA
Session Introduction
Stevens M Brumbley
University of North Texas, USA
Title: Metabolic engineering of a high biomass C4 grass for commercial scale production of bioplastics
Time : 12:30-13:00
Biography:
Stevens M Brumbley completed his PhD at the University of Georgia in 1992 and postdoctoral studies at the University of California Riverside in 1993. He worked in the Australian Sugarcane Industry from 1993 to 2006 and at the University of Queensland’s Australian Institute for Bioengineering and Nanotechnology for 2006 to 2011. He is currently a tenured Associate Professor of Plant Metabolic Engineering at the University of North Texas. He has published more than 50 papers in high impact journals.
Abstract:
Sugarcane is an important high biomass crop grown throughout the tropical and subtropical regions of the world. In Brazil, sugarcane has a central role in not only the production of sucrose but also in the production of biofuels and bio-based chemicals. In the future, these bio-based chemicals may be derived not only from bio-refineries but also from genetically engineered sugarcane itself. We have been metabolically engineering sugarcane to function as a bio-factory producing bio-plastics and bio-plastic precursors. Sugarcane was metabolically engineered to produce polyhydroxybutyrate (PHB), a member of the class of biopolymers known as polyhydroxyalkanoates (PHA). Most of our work has focused on PHB using a three enzyme pathway from a soil bacterium (Ralstoniaeutropha) [β-ketothiolase (PhaA), acetoacetyl-reductase (PhaB) and PHB synthase (PhaC)]. Transgenes were expressed in the nucleus of sugarcane and the enzymes were targeted to plastids. In plastids, using a maize polyubiquitin promoter, PHB granules were detected in all cell types in sugarcane stalks and leaves, however, the highest accumulation was in bundle sheath cells. A number of strategies were used to increase the levels of PHA accumulation in sugarcane leaf tissue. An early detection system was developed using Nile Blue. A staining and fluorescent microscopy showing that PHA could be detected in leaf tissue from tissue culture plantlets regenerated from the transgenic callus. When a stronger promoter was used (maize chlorophyll A/B binding protein promoter) PHA accumulation was significantly higher in leaf tissue of sugarcane. Knocking out competition for acetyl-CoA in sugarcane resulted in a 50% increase in PHB levels in leaves. Replacing PhaA with a novel enzyme resulted in another 4 fold increase in PHA levels resulting in leaves producing commercial levels of PHA. In addition to plastid production, PHB and PHB/PHA copolymers were engineered into sugarcane peroxisomes by targeting the PhaA, PhaB, and either PhaC or the Pseudomonas aeruginosa PhaC1 to peroxisomes. We found both PHB and PHB/PHA copolymer granules localized in peroxisomes and vacuoles in sugarcane leaves. The biopolymers we are producing have very high molecular weight with the highest at close to two million daltons. The highest production level of PHA polymer recorded to date in sugarcane is 12% of the total leaf dry weight.
Soumendra Rana
Indian Institute of Technology Bhubaneswar, India
Title: Structural modeling of C5a receptor: Molecular insights into agonism and antagonism
Time : 14:00-14:30
Biography:
Soumendra Rana has completed his PhD at the age of 29 years from IIT Bombay and postdoctoral studies from Washington University School of Medicine in St. Louis, including University of Arizona, Tucson. He is currently an assistant professor (Chemistry and Biosciences) at the IIT Bhubaneswar, one among the 14 premier technical institutes of the country. He has published more than 12 papers in reputed journals and has been serving as an editorial advisory board member in International Journal of Chemical and Pharmaceutical Review and Research.
Abstract:
The complement component fragment 5a receptor (C5aR), also known as the anaphylatoxin receptor is one of the two major G-protein coupled receptor (GPCR) that demonstrates a high affinity interaction with C5a, the most potent pro-inflammatory polypeptide of the complement system, known for its pleiotropic effects in both immune and non-immune cells. No wonder, the C5a-C5aR interaction has been tagged druggable for discovery of targeted therapeutics.1 However, the molecular basis of agonism or antagonism in C5aR is yet to be established clearly, largely due to the unavailability of a structure of C5aR. In addition, while the role of allosterism in C5aR is discussed in the literature, it is completely unheard for C5a, a potential drug target that modulates the downstream signaling of C5aR. It not only hinders the discovery and development of new lead molecules, but also affects the rational optimization of the known lead molecules as potential therapeutics, targeting the C5a-C5aR signaling axes. In our quest toward better understanding of C5a-C5aR interaction, 2 we have recently generated the first set of atomistic model structures of inactive and meta-active C5aR in excellent agreement with the previously reported binding and signaling studies. 3 Further, we have also identified a pair of “allosteric switches”4 on C5a that potentially modulate the C5aR signaling. It is noteworthy that the peptide agonist, C5a and the small molecule antagonist5 NDT demonstrate binding at the exact same site on the meta-active structure of C5aR with distinctly different binding modes.6 In summary, the inactive, agonist and antagonist bound meta-active C5aR structures provide important structural insights, previously not known at an atomistic resolution, in regard to the ligand binding sites, selectivity and activation of C5aR, which will be discussed in detail.
Xiaolian Gao
University of Houston, Houston TX
Title: Mass production of high fidelity oligonucleotides as feed stock of post-genome-sequencing synthetic biology (PGS-SB)
Time : 14:30-15:00
Biography:
Xiaolian Gao co-founded VisiGen Biotechnologies, Inc., in 2000. She is Professor of Chemistry, Biology and Biochemistry and Director of the Keck/IMD NMR Center at the University of Houston. After spending more than two years working at then Glaxo Pharmaceuticals as a Principle Research Investigator in structural biology, she joined the Department of Chemistry at the University of Houston. At the Keck NMR Center that she established at the University, she has collaborated with many groups in Houston and nationwide. Her group has discovered and published unique structures of biologically important DNA and ligand-DNA complexes. Pursuing research in the interdisciplinary areas of chemistry and biology, she developed novel methods for massively parallel synthesis of biomolecule and chemical microarrays, which became the founding technology of Xeotron Co. Ms. Gao holds a BS degree from the Beijing Institute of Chemical Engineering and PhD in Chemistry at Rutgers University. During her postdoctoral work in NMR-based structure biology at the medical school of Columbia University in late 80’s she was among the first few who solved the structures of several important Ligand-DNA complexes using high resolution nuclear magnetic resonance (NMR) spectroscopy.
Abstract:
Rapid progress in genome sequencing and functional genomics has set up the stage for a new era of engineering-based postgenome- sequencing synthetic biology (PGS-SB). As the blueprints, i.e., the DNA coding sequences, of biological devices of specific functions are largely known, engineering these devices, further bio-systems which are made of designed devices are tacks of construction of structures of hundreds and thousands of nucleotides in defined orders and contents. Oligonucleotides thus become the fundamental building blocks of aforementioned bio-devices and systems. We report a micro-fluid microarray chip technology platform, for programmable engineering oligonucleotides in quality and quantity which coupled with a seamless workflow for generation of high fidelity oligonucleotides, we will demonstrate the application of these polynucleotides in a varieties of PGS-SB applications. Our goal was to offer affordable solutions to general laboratories interested in PGS-SB.
Pushpa Agrawal
R. V. College of Engineering, India
Title: Development of green formulation for dual purpose of disease management and plant development
Time : 15:00-15:30
Biography:
Pushpa Agrawal has completed her PhD degree in the year 1984 from DAVV, Indore and Post-doctoral studies from Miami University, USA and India Institute of Science Bangalore, India. She is a Professor of Biotechnology and Dean, Post-graduate studies in Biotech and Chemical Engineering at R. V. College of Engineering, Bangalore India. Many scholars are pursuing and completed PhD under her guidance. She has presented over 100 research papers in conferences and published more than 40 papers in reputed journals. She has co-authored a book on biotechnology and chapters in 2 books and has been serving as Managing Editor and Editorial Board Member of reputed journals. She has also received various awards, honors and fellowships.
Abstract:
The use of chemical fertilizers and pesticides in commercial farming gives the threat of gradual aggravation of soil fertility. Use of agriculturally important microorganisms in different combinations is the only solution for restoration of soils. The bio-formulations using humic acid with suitable bio-control microorganisms namely, Pseudomonas fluorescens and Trichoderma harzianum have been developed to replace chemical fertilizers. Trichoderma harzianum is a major bio-control agent against a wide range of phyto-pathogenic organism of economically important crops and extensively used in various parts of the world for plant disease management. These bio-control agents are also known to degrade complex organic molecules into simpler molecules which help to improve soil fertility. Humic acid is an organic fertilizer which enhances the plant growth. The bio-organic formulations have been developed for controlling or suppressing of fungal diseases (Fusarium species) as well as to increase the plant growth and yield by increasing the soil fertility in an eco-friendly manner. The present report reveals an additive effect of microorganism with humic acid in both disease management and better plant growth. The results revealed that the Trichoderma harzianum with 2% humic acid based formulation was effective in inhibiting Fusarium growth in vitro. The liquid bio-formulation of humic acid along with the Pseudomonas fluorescens was tested and compared for viability as well as its inhibitory characteristics against Fusarium oxysporum, a fungus which cause wilt of tomato. Cell viability tests were carried out for the bio-formulations by plate count method. Field studies were conducted for two crop varieties-radish and tomato. The results of in vivo and in vitro studies revealed better support for the viable cells as well as leafing and fruit
- Posters
Location: Gallery
Session Introduction
Gustavo Stadthagen
BIOASTER, France
Title: Controlled ultrasound cavitation for DNA delivery into bacteria and yeast
Biography:
Gustavo Stadthagen holds PhD in Microbiology from the University of Paris Denis Diderot. He has published more than 10 articles on Molecular Genetics and Biochemistry of Infectious Agents, Cancer Cells and Biotechnology Relevant Microorganisms. He currently works as a Research Scientist at the Unit of Protein and Expression System Engineering of BIOASTER, a new technology research institute on infectiology and microbiology. His research using an integrated approach that includes enhanced recombinant DNA construct design, assembly, delivery and stabilization aims to develop improved protein expression systems for biopharma and biotech applications.
Abstract:
Sonoporation is an ultrasonic cavitation-dependent method for gene delivery, mostly applied to higher eukaryotic cells and tissues. Cavitation in the vicinity of cells induces pore formation in the cell envelope and mechanical internalization of extracellular DNA. Its application to biotechnology useful organisms is less developed. Here we evaluated the CaviBoxR sonoporation device for plasmid delivery into the yeast Kluyveromyces lactis and the bacterium Escherichia coli BL21, commonly employed organisms for bio-production. CaviBoxR tightly controls cavitation through combination of multiple ultrasound beams under strong negative pressures. We evaluated the effect of different cavitation conditions (intensity, duration, duty cycle) and cell suspension media composition on cell viability and functional plasmid uptake. Optimal DNA delivery into K. lactis kept in YPD culture medium and incubated with linearized plasmid before sonoporation was achieved at CI 14 for 60 seconds with minimal effects on cell viability. Transformation efficiency was comparable to conventional chemical transformation of this host. Plasmid uptake by E. coli occurred in LB culture medium supplemented with 100 mM CaCl2 at CI 12-14 for 20 seconds. Under the same cavitation conditions, transformation was 100 times more efficient when bacteria were suspended in a solution of 25 mM MgCl2, 100 mM CaCl2 and 10% Glycerol. These first results demonstrate the feasibility of DNA delivery into bacteria and yeast species of biotechnological interest by sonoporation, directly in culture medium with minimal manipulation. Improvement of this method holds the potential of simple parallel transformation of large strain collections and or recombinant DNA libraries.
Richard McLean
University of Lethbridge, Alberta, Canada
Title: Mitigation of poultry-borne Campylobacteriosis by an engineered enteric commensal
Biography:
Richard McLean is a master student at University of Lethbridge Canada. McLean contributed to a ground-breaking study that was recently published in Nature, the world’s most-cited interdisciplinary science journal with work he accomplished as an undergraduate student in an Applied Study setting with Dr. Wade Abbott, an adjunct chemistry and biochemistry professor.
Abstract:
Campylobacter jejuni is the leading cause of human gastroenteritis in the developed world with cases in of Campylobacteriosis in Canada numbering nearly 50% of all bacterial foodborne illness. Primary infections are typically self-limiting however numerous secondary sequelae can develop including reactive arthritis, irritable bowel disorder, inflammatory bowel disease, and Guillain-Barre syndrome, the leading cause of acute fl accid paralysis in North America. Because as much as 70% of human Campylobacteriosis can be traced to the consumption of contaminated poultry, this project aims to prevent C. jejuni from proliferating in the chicken gut. Numerous approaches have been attempted previously including the addition of bacteriophages or bacteriocins to the feed, chicken vaccination, positive selection of C. jejuni free birds and a variety of methods to limit physical exposure of the birds to the bacterium. This project uses a synthetic biology approach to engineer the human gut commensal Bacteroides thetaiotaomicron to produce and secrete Nano-bodies within the chicken gut. Nano-bodies were raised against C. jejuni flagella and the focus of this project was to establish adequate expression levels, integrate required genes into the genome of B. theta and demonstrate the secretion of functional protein. The benefit of this approach will be the continued production of Campylobacter-active compounds within the chicken gut, which would be an economic and technological advancement over feed supplementation. In addition, this approach will enable the rational selection of therapeutic targets to help avoid the development of resistance.
Zehra Tatli
Hacettepe University, Turkey
Title: Comparison of intracellular and extracellular cellulase production by recombinant bacterium Escherichia coli
Biography:
Zehra Tatli is a Master of Science student in Bioengineering at Hacettepe University, Turkey and she has Bachelor’s of Science degree in Biology. Currently, she is working on her MSc thesis titled “Novel cellulase enzyme production towards biofuels sector by recombinant bacterium Escherichia coli” supported by TUBITAK (The Scientific and Technological Research Council of Turkey). She is interested in Synthetic Biology and wants to improve herself further during PhD studies on recombinant protein biogenesis in microorganisms and on the development of platform technologies for protein expression and engineering.
Abstract:
Low cost, pH and thermo-stable cellulase enzymes are an important factor for commercially viable production of bioethanol which is a renewable source of energy. Nowadays, the cost of cellulase accounts for 40-50% of the total ethanol production cost and it is targeted to be reduced 5-folds for commercial efficiency. Instead of food raw materials such as corn and sugar cane, ethanol obtained from cellulosic biomass by endoglucanase type of cellulase will reduce production costs. In this process, the choice of the host cell is extremely important in order to develop more economical production processes. Escherichia coli bacterium is one of the most preferred hosts for the production of recombinant proteins. On the other hand, enzymes produced in bacterial systems are known to be more economical compared to eukaryotic cells provided that they can be secreted in high amounts. In this context, recently codon optimized novel Cel5A enzyme was expressed intracellularly and extracellularly in E. coli and bioprocess optimization studies were followed by Western blot and spectrophotometric enzyme activity assays. We increased intracellular enzyme activity 50-fold up to 0.74 IU/mL and extracellular enzyme activity 5-fold up to 1.5 IU/mL. The recombinant cellulase enzyme and the bioprocess developed in this study have vital importance for overcoming the
Neema K N
Sri Jayachamarajendra College of Engineering, India
Title: In silico molecular docking of ligands for inhibition of L-amino acid oxidase from Calloselasma rhodostoma
Biography:
Neema K N is a Research scholar in Department of Biotechnology, Sri Jayachamarajendra College of Engineering. She is currently pursuing her PhD from JSS Research Foundation in affiliation with University of Mysore. She has been awarded with Junior Research Fellowship, UGC, Government of India. She has published a paper in reputed journal and has presented poster in 2nd National Conference of Toxicological Society of India.
Abstract:
L-amino acid oxidases (LAAO; EC 1.4.3.2) are flavo-enzymes catalyzing the oxidative deamination of L-amino acids and produces reactive oxygen species (ROS) including Hydrogen peroxide. H2O2 mediates in the process of cell death by necrosis, platelet aggregation, apoptosis etc. Several biological activities of LAAOs from snakes have been reported including apoptosis-inducing activity. Development of a lead with the combination of biological activity and drug like properties is needed. Both features can be estimated primarily by in silico virtual docking strategies in drug discovery and development. In this study, the crystal structure of L-amino acid oxidase 2IID was imported into Maestro 9.3 following subsequent removal of all crystallographic water molecules from the PDB. Bio-actives from several medicinal plants were screened for the inhibition of LAAO from Calloselasma rhodostoma. Th e possibility of binding, precise location of binding sites and mode of binding ligand was carried out using automated docking soft ware Glide XP. The efficacy was evaluated based on scoring function employed in Glide XP (Schrodinger platform). The promising compounds were employed for ADME/Tox studies. Our result showed that Limocitrin bioactive from Citruslimon completely buried deeply inside the catalytic tunnel, blocking the transfer of hydride ion at isoalloxazine ring resulting in obstruction for formation of H2O2, thereby inhibiting LAAO. Furthermore pharmacophore modeling suggests the better binding in presence of hydroxyl moieties.
Shalini Koshle
Sri Jayachamarajendra College of Engineering, India
Title: Screening of phytochemicals for the inhibition of JAK2 by molecular docking
Biography:
Shalini Koshle is a Senior Research Fellow in Department of Biotechnology, Sri Jayachamarajendra College of Engineering. She is currently pursuing PhD from Visvesvaraya Technological University, India. She has been awarded with Rajiv Gandhi National Fellowship, University Grant Commission, Government of India. She has published two papers in reputed journal and given oral presentation National Conference on “Technological Advancements in Chemical and Environmental engineering”.
Abstract:
Janus kinases-2 (JAK2) is a member of the Janus Kinase family and has been implicated in signaling by members of the type II cytokine receptor family. JAK2 is involved in the regulation of various pathways of cell growth, development, differentiation or histone modifications. Inhibition of JAK down-regulates phosphorylation of transcription factors known as STAT and blocks the JAK-STAT signaling pathway which is vital for tumorigenesis. In the present study molecular docking was carried out to elucidate the interaction of phytochemicals with Protein Kinase 2 domain for the inhibition of JAK2 to block cytokine signaling. PDB ID code 4C61 from the Protein Data Bank was used to retrieve JAK2 crystal structure at 2.45- A resolution and utilized for molecular docking under Schrodinger platform. Phytochemical, demethoxycurcumin from Curcuma longa as JAK2 inhibitor to modulate the JAK/STAT pathway. It is found that the phytochemical is potent to block the access at the site of Protein Kinase 2 domain. The efficacy was evaluated based on scoring function employed in Glide XP. Further ADME/ Tox studies validate the efficacy of the phytochemical to possess druggable like characteristics pertaining to standard drugs. Pharmacophore modeling of demethoxycurcumin showed binding energy with substitution of different R-groups.