Day 2 :
University of Guelph, Canada
Keynote: Production of antimicrobial peptide, protegrin-1 in Pichia pastoris and its protection against DSSinduced colitis
Time : 10:00-10:25
Julang Li is a Professor at the Department of Animal Bio-Science, University of Guelph, Canada. Her research program balances both the fundamental and applicable aspects of animal and biomedical science. Her fundamental research focuses on studying the regulation of follicular development and oocyte maturation. The more napplicable research program in her laboratory aims to enhance animal and human health via biotechnological approaches. In this regard, they have been using food grade bacteria as a bioreactor and vehicle to produce and deliver recombinant proteins such EGF and antimicrobial peptide to the intestine for enhancing early weaned pig development.
Bacterial resistance to conventional antibiotics is a growing public health concern. One of the means to cope with this issue is to use antibiotic alternatives. Protegrin-1 (PG-1) is a porcine cathelicidin Antimicrobial Peptide (AMP) that can exert its activity against a broad range of microorganisms including bacteria and enveloped viruses, allowing it to be an attractive candidate for therapeutic use. In addition, as the peptide is an integral part of the innate immune system, protegrins may have other functions such as immune stimulating or modulating effect, similar to its human cathelicidin counterpart. Little is known about the involvement of porcine cathelicidin in this regard. We have generated codon-optimized proform PG-1 and mature PG-for expression in Pichia pastoris yeast. In this study, we further investigated the potential inflammatory modulating and protective role of PG-1 in well-established Dextran Sulfate Sodium (DSS)-induced colitis model in mice. Body weight, clinical symptoms,histology and gene expression of colonic tissues were assessed. Relative expression of inflammatory cytokines (COX-2 and TNF-a) was significantly reduced in protegrin treatment groups (p<0.05). Protegrin treatment prevented significant body weight loss and improved Disease Activity Index (DIA) scoring (p<0.05) compared to the untreated DSS-control mice. Histological analyses indicate reduced mucosal erosion and sub-mucosa inflammation in protegrin-treated groups. Overall, oral administration nof protegrin was demonstrated to be protective against colitis induction in mice. Histological and gene expression results are reflective of the phenotype observed in protegrin DSS-treated mice and the controls. Resulting data establishes the potential use of protegrins to modulate intestinal health in vivo.
- Environmental BiotechnologyBiotechnology in Healthcare IndustryIndustrial Microbiology and Enzyme Technology
University of Guelph, Canada
T. B. Karegoudar
Gulbarga University, India
National Institute of Pharmaceutical Education and Research, India
Title: Improving organophosphate-hydrolyzing activity of human paraoxonase 1 by mutagenesis approach
Time : 10:25-10:50
Rajan K Tripathy has completed his MSc degree from Annamalai University, Chidambaram (India) and is currently pursuing Doctoral studies from National Institute of Pharmaceutical Education and Research (India). He is also an employee of this organization. He has published more than 10 papers in reputed journals and is an inventor in 2 patents.
Organophosphates OPs) are extremely toxic compounds that are commonly used as pesticides and also as chemical warfare nerve agents. OP compounds irreversibly inhibit neurotransmitter metabolizing enzyme, acetylcholinesterase (AChE). Inhibition of AChE results in severe cholinergic crisis caused by increased concentration of acetylcholine at the cholinergic nerve–nerve and nerve–muscle synapses. Some of the examples of toxic OPs are pesticides and chemical warfare nerve agents like malathion, chlorpyrifos-oxon, parathion, soman, sarin, etc. The current treatment of OP-poisoning includes combination of anti-cholinergic drugs together with quaternary oxime reactivators which is not much effective. Human paraoxonase 1 (hPON1), a 40 kDa enzyme predominantly found associated with HDL in the blood circulation, can potentially hydrolyze numerous OPs and is thus considered as a promising catalytic scavenger candidate for the therapy of OP-poisoning. However, the OP-hydrolyzing capacity of native hPON1 is low. In an attempt to increase the OP-hydrolyzing capacity, random mutagenesis of recombinant human PON1 (rh-PON1) was carried out by both XL-1 Red mutator stain and error-prone PCR methods for two rounds of molecular evolution. The protein was expressed, purified and characterized. Some of the mutant in the second round of evolution showed 20-30 folds increase in OP-hydrolyzing activity. These novel mutants can be of therapeutic use against OP-poisoning.
Gulbarga University, India
Title: Purification and characterization of amidase from Paracoccus sp. SKG: Utilization of amidase inhibited whole cells for bioconversion of acrylonitrile to acrylamide
Time : 11:10-12:10
T.B.Karegoudar, Professor of Biochemistry, working in the university for the past 28 years. He has focused his research on bioremediation of xenobiotics, microbial bioconversions and isolation of industrially important microbial enzymes and their applications. He has isolated and characterized several potential strains and deposited in the national cultural collections. He has successfully completed major projects funded by many national funding agencies. He has published more than 65 research publications in reputed journals. He has successfully guided 13 PhD students. He served as Chairman, Department of Biochemistry of Gulbarga University. He is in the review panel of several microbiology and biotechnology journals. He is an editorial member of few scientific journals.
A bacterial strain Paracoccus sp. SKG capable of utilizing aliphatic nitriles as a sole source of carbon and nitrogen was isolated. Degradation of nitriles follows the bi-enzymatic pathway with the successive action of nitrile hydratase (NHase) and amidase. The inducible amidase from cell-free extract of Paracoccus sp. SKG was purified and characterized. The amidase was purified to 93.0 fold with a recovery of 46.5% yield and an estimated molecular mass of 90 kDa. The purified amidase exhibits highest activity at pH 7.5 and temperature at 50oC. This enzyme is highly specific to aliphatic amides with highest activity to acetamide, propionamide and acrylamide, but not to aromatic amides. Among the metal ions studied, Mg2+, Mn2+ and Ni2+ ions increase the enzyme activity, whereas Cu2+ completely inhibited the amidase activity. In vitro inhibition of amidase was confirmed by preincubation of the whole cells Paracoccus sp. SKG with Cu2+. The use of amidase inhibited whole cells of Paracoccus sp. SKG was demonstrated as a biocatalyst for the conversion of acrylonitrile to acrylamide in a batch reaction. The bioconversion of acrylonitrile to acrylamide resulted in 27g/L of acrylamide with 65% of conversion under optimal conditions of pH (7.0) and temperature (30oC).
University of Reading, UK
Time : 12:10-13:10
Vimal Karani S is a Lecturer in Nutrigenetics at the University of Reading, UK. He did his Post-doctoral training at the MRC Epidemiology unit (Cambridge, UK) and University College London (UK). He has an interdisciplinary academic background, with qualifications from Medical Genetics, Bioinformatics, Molecular Biology and Genetic Epidemiology. His primary research interests focus on the investigation of gene-nutrient interactions on metabolic- and CVD-related outcomes using combined approaches from genetic epidemiology, statistical genetics and molecular biology. His long term goal is to use the findings from observational studies to carry out human intervention studies with a view towards developing industrial collaborations to facilitate ‘Personalized Nutrition’..
The concept of “Personalized” Medicine is now being extended to the field of Nutrigenetics, which investigates the impact of gene variation responses to intake of different nutrients. The ability of Nutri-genetics to determine what nutrients will produce the desired impact on metabolic balance (as influenced by individual genetic make-up) is at the core of personalized nutrition. Obesity is a heritable trait that arises from the interactions between multiple genes and lifestyle factors such as diet and physical inactivity. Dietary factors play an important role in the development of obesity because of the variation in the food that is being consumed in different parts of the world. Although several studies have examined the gene x nutrient interactions, the findings have been quite inconsistent and hence, unable to develop an optimum diet for each ancestral population. Some of the challenges in performing nutrigenetics research are 1) genetic heterogeneity, 2) lack of understanding of the metabolic pathways and 3) insufficient sample size. With genome-wide association study (GWAS) data now available on numerous large cohorts, it has become possible to embed candidate gene studies within GWASs, testing for association on a much larger number of candidate genes than previously possible. The talk will highlight three main aspects: 1) why do we do gene-diet interaction analysis? – Findings from DiOGenes study, 2) why large samples are required to conduct genetic epidemiological studies? –Findings from D-CarDia Collaboration and 3) Nutrigenetics in developing countries – Findings from GeNuIne Collaboration
University of Calabar, Nigeria
Title: Process optimization of bio-ethanol production from cassava peels using different microbial innoculants
Time : 13:50-14:15
Edak Aniedi Uyoh is a Professor Department of Genetics and Biotechnology, University of Calabar, Nigeria.
Cassava peel is one of the major biomass wastes in Nigeria, obtained from processing of cassava tuber for human consumption, starch production and industrial uses. A few reports are available of its potential in bio-ethanol production which is a promising alternative energy source for crude oil. In this study, an attempt was made to optimize the production of bio-ethanol using different microbial inoculum for the simultaneous saccharification and fermentation of cassava peels in three cassava varieties at an optimal pH of 5.0 and a temperature of 28oC in 21 days. The cultures were distilled on the 7th, 14th and 21st days. The combinations of inoculum used were: A=Rhizopus nigricans+Saccharomyces cerevisae; B=Aspergillus niger+Saccharomyces cerevisae; C=Rhizopus nigricans+Aspergillus niger+Saccharomyces cerevisae; D=Rhizopus nigricans+Spirogyra africana+Saccharomyces cerevisae; E=Aspergillus niger+Spirogyra africana+Saccharomyces cerevisae and Control=Saccharomyces cerevisae. Results obtained showed significant differences (P<0.05) in the amount and concentration of ethanol produced among the five innoculants but no significant difference (P>0.05) among the three cassava varieties. Cassava peels from variety TME 4779 gave the highest amount of ethanol with a yield of 19.72 g/cm3 when pH of the culture was optimized using Rhizopus nigricans+Spirogyra africana+Saccharomyces cerevisae on the 7th day. It gave ethanol yield of 18 g/cm3 when pH of the culture was optimized on the 21st day but gave 13.33 g/cm3 when the pH of the culture conditions was left in its natural state. The lowest amount of ethanol yield obtained were 4.17 g/cm3, 5.28 g/cm3 and 6.66 g/cm3 from the cassava peels of TME 419, TME 0505 and TME 4779 respectively in an optimized condition using Saccharomyces cerevisae alone. The implications of these findings in optimizing production of bio-ethanol from cassava peels and the prospects for the petroleum industry are highlighted.
King Abdulaziz City for Science and Technology, Saudi Arabia
Time : 14:15-14:40
Sami A Al Yahya is a Professor at King Abdulaziz City for Science and Technology, Saudi Arabia and a Honorary lecturer.
The aim of the work reported was to investigate the role of a range of micro-organisms in the functioning of the environment, particularly in relation to the biogeochemical cycles. At this point, a study was made of the microbial activity of phosphate soils in comparison with that displayed by a fertile agricultural soil. The microbial transformations studied included phosphate solubilisation, the oxidation of elemental sulphur to sulphate, nitrification and the hydrolysis of urea. These processes were collectively used to study the biogeochemical activity of soils. Bacterial population densities in the soils, fertile soils and phosphate soil, desert vegetated soil samples were also determined. A variety of bacterial isolates from phosphate soils, desert vegetated soil and desert non-vegetated soil samples have been identified using molecular identification techniques like DNA extraction, PCR amplification, determinations of 16SrRNA gene sequences.
Tallinn University of Technology, Estonia
Time : 14:40-15:05
Kristjan Piirimäe, PhD, studies as a researcher on surface water quality management in Tallinn University of Technology, Estonia. He also teaches international master courses on eco-design and environmental impact assessment. Piirimäe has studied microbiology, hydrobiology and GIS modelling. He participates in a EU-funded BONUS programme project “Cost efficient algal cultivation systems – A source of emission control and industrial development (Microalgae)”. Piirimäe has worked on spatial planning of biogas stations and decision support in environmental management. He has worked on large-scale quantification of nutrient pollution, wastewater amounts and biogas sources
The EU aims to reduce dependence on fossil fuels. One possible mean for that might be high rate algal ponds (HRAPs) which photosynthesize biofuel, e.g. algae. HRAPs might effectively replace fossil fuels in future if economic viability is achieved. Today, the only identified economically viable solution for HRAP is wastewater treatment. The EU Urban Waste Water Directive, in turn, has set strict requirements for N and P concentration in wastewater effluent. These requirements remain challenging for many settlements. We studied the potential of HRAPs to meet both waste water and energy targets in Estonia, a EU member state. We identified pollutants limiting the achievement of water quality requirement while assessing also the effluent conditions for the growth of microalgae in HRAPs. Of 979 wastewater outlets, 56% indicated P concentration higher than 2 mg/l, a norm for most settlements while N concentration exceeded the norm of 15 mg/l in 35% of the outlets. Hence, the main problem was pollution with P. Considering that algal cells may contain, depending on conditions, N/P molar ratio between 4:1 and 40:1, in total 78% of all outlets served suitable nutrient conditions for algal growth. In total 8% of analyzed outlets had critical N deficit while 15% showed P deficit. Considering both selection criteria – high N or P content as well as suitable N/P ratio – in total 51% of all outlets matched for the possible treatment with HRAP. However, of the total Estonian consumption of solid fuels, such wastewater-fed HRAPs cannot theoretically cover more than 0.06%. We conclude that while they might possibly contribute towards better water quality, they cannot significantly reduce the demand for fossil fuels.