Ashok Pandey, Ph.D.
Scientist F & Head, Biotechnology Division, National Institute for Interdisciplinary Science and Technology-CSIR), Thiruvananthapuram, India
Alternative renewable resources: Issues and perspectives for India – the case of transport fuels
With the increase in the urbanization way of life and also more and more dependence on materialistic life, there is substantial growing demand for the energy. The science and technological policy of the India has looked several avenues to fulfill this demand through alternative resources such as solar energy, wind energy, tidal energy, bioenergy, etc. The demand for the transport sector is largely met through the import (~70%). Biofuels, in particular bioethanol from lignocellulosic biomass offer attractive possibilities in this regard.
The sugar platform which generates ethanol is considered to be the most valuable solution to the transport fuel demand. Bioethanol can be generated from grains as well as from lignocellulosic plant material by their saccharification to sugars and subsequent fermentation of the sugars to produce ethanol. Bio-ethanol as a transportation fuel is attractive since it is more energy efficient than gasoline and produces less emissions. The benefits of developing biomass to ethanol technology(s) include: increased national energy security, reduction in GHG emissions, use of renewable resources, economic benefits and creation of employment and the foundation of a carbohydrate based chemical industry. However, the utilization of lignocellulosic biomass for fuel generation has not been given the sort of attention it ought to receive. It is known that the technology for ethanol production from biomass has to evolve greatly for an economical commercial scale utilization of the renewable biomass resources. Biomass requires extensive processing involving multiple steps for hydrolysis and fermentation of the raw material for producing ethanol. Feed stock availability, pretreatment, saccharification, fermentation and ethanol recovery are all factors which influence the production of ethanol and which needs R&D efforts for overall improvement of the production economics.
Bioconversion of lignocellulosic biomass (LB) can contribute significantly to the production of organic chemicals also. LB is also considered to be the only foreseeable source of energy. LB is mainly composed of (dry wt basis): cellulose, 40-60; hemicellulose, 20-40; and lignin, 10-25%. Most efficient method of biomass hydrolysis is through enzymatic saccharification5 using cellulases and hemicellulases. Fungal cellulases (FCs) have proved to be a better candidate than other microbial cellulases, with their secreted free cellulase complexes comprising all three components of cellulase [endoglucanases, exoglucanases and cellobiases (glucosidases).
The Centre for Biofuels at NIIST, Trivandrum, India aims ultimately to develop technologies and processes which will address the nation’s need for making fuel ethanol from the renewable resource: biomass. It is proposed to direct R&D activities at the major requirements of a biomass-ethanol technology, which include production of cellulases, hydrolysis of biomass, and ethanol fermentation. Viable technologies for each of these processes will contribute to the overall process development for fuel alcohol production from cheap and renewable biomass resources.
The lecture would present perspectives on bioethanol from lignocellulosic feedstocks.
References
- Biofuels- Alternative Feedstocks and Conversion Processes, Editors- Ashok Pandey, C Larroche, SC Ricke, CG Dussap & E Gnansounou, Academic Press, Elsevier Inc; San Diego, USA, p629 (2011) ISBN: 978-0-12-385099-7
- Handbook of Plant-Based Biofuels, Editor- Ashok Pandey, CRC Press, Francis & Taylors, Boca Raton, USA, p 297 (2008) ISBN 978-q-5602-2175-3
- Biofuels II, Special issue of Journal of Scientific & Industrial Research, Guest Editors- E Gnansounou, C Larroche and Ashok Pandey, 67(11), 837-1040 (2008) ISSN: 0022-4456
- Biofuels, Special issue of Journal of Scientific & Industrial Research, Guest Editors- C Larroche and Ashok Pandey, 64(11), 797-988 (2005) ISSN: 0022-4456
Bharat B. Chattoo, Ph.D.
Professor, Faculty of Science M.S.University of Baroda, India
Biology of plant infection by Magnaporthe oryzae
The rice blast disease caused by the ascomycetous fungus Magnaporthe oryzae is a major constraint in rice production. Rice-M.oryzae is also emerging as a good model patho-system to investigate how the fungus invades and propagates within the host. Identification and characterisation of genes critical for fungal pathogenesis provides opportunities to explore their use as possible targets for development of strategies for combating fungal infection and to better understand the complex process of host-pathogen interaction.
We have used insertional mutagenesis and RNAi based approaches to identify pathogenesis related genes in this fungus. A large number of mutants were isolated using Agrobacterium tumefaciens mediated transformation (ATMT). Characterisation of several interesting mutants is in progress. We have identified a novel gene, MGA1, required for the development of appressoria. The mutant mga1 is unable to infect and is impaired in glycogen and lipid mobilization required for appressorium development. The glycerol content in the mycelia of the mutant was significantly lower as compared to wild type and it was unable to tolerate hyperosmotic stress. A novel ABC transporter was identified in this fungus. The abc4 mutant did not form functional appressoria, was non-pathogenic and showed increased sensitivity to certain antifungal molecules implying the role of ABC4 in multidrug resistance (MDR). Another mutant MoSUMO (MGG_05737) was isolated using a Split Marker technique; the mutant showed defects in growth, germination and infection. Immuno-fluorescence microscopy revealed that MoSumo is localized to septa in mycelia and nucleus as well as septa in spores. Two Dimensional Gel Electrophoresis showed differences in patterns of protein expression between Wild Type B157 and MoΔSumo mutant. We also isolated and charaterised mutants in MoALR2 (MGG_08843) and MoMNR2 (MGG_09884). Our results indicate that both MoALR2 and MoMNR2 are Mg2+ transporters, and the reduction in the levels of CorA transporters caused defects in surface hydrophobicity, cell wall stress tolerance, sporulation, appressorium formation and infection are mediated through changes in the key signaling cascades in the knock-down transformants. (Work supported by the Department of Biotechnology, Government of India)
![Invited Talk: Remote Patient Monitoring – Challenges and Opportunities @ Amriteshwari Hall | Vallikavu | Kerala | India](http://www.amritabioquest.org/conference/2013/wp-content/uploads/sites/2/2015/02/biomed.jpg)
Jaydeep Unni, Ph.D.
Sr. Project Manager, Robert Bosch Healthcare Systems, Palo Alto, CA
Remote Patient Monitoring – Challenges and Opportunities
Remote Patient Monitoring (RPM) is gaining importance and acceptance with rising number of chronic disease conditions and with increase in the aging population. As instances of Heart diseases, Diabetes etc are increasing the demand for these technologies are increasing. RPM devices typically collect patient vital sign data and in some case also patient responses to health related questions. Thus collected data is then transmitted through various modalities (wireless/Bluetooth/cellular) to Hospitals/Doctor’s office for clinical evaluation. With these solutions Doctors are able to access patient’s vital data ‘any time any where’ thus enabling them to intervene on a timely and effective manner. For older adult population chronic disease management, post-acute care management and safety monitoring are areas were RPM finds application. That said, there are significant challenges in adoption of Remote Patient Monitoring including patient willingness and compliance for adoption, affordability, availability of simpler/smarter technology to mention a few. But experts contend that if implemented correctly Remote Patient Monitoring can contain healthcare expenditure by reducing avoidable hospitalization while greatly improving quality of care.
![Delegate Talk: Inflammation Induced Epigenetic Changes in Endothelial Cells: Role in Vascular Insulin Resistance @ Acharya Hall | Vallikavu | Kerala | India](http://www.amritabioquest.org/conference/2013/wp-content/uploads/sites/2/2015/02/gtm.jpg)
Aswath Balakrishnan, Kapaettu Satyamoorthy and Manjunath B Joshi
Introduction
Insulin resistance is a hall mark of metabolic disorders such as diabetes. Reduced insulin response in vasculature leads to disruption of IR/Akt/eNOS signaling pathway resulting in vasoconstriction and subsequently to cardiovascular diseases. Recent studies have demonstrated that inflammatory regulator interleukin-6 (IL-6), as one of the potential mediators that can link chronic inflammation with insulin resistance. Accumulating evidences suggest a significant role of epigenetic mechanisms such as DNA methylation in progression of metabolic disorders. Hence the present study aimed to understand the role of epigenetic mechanisms involved during IL-6 induced vascular insulin resistance and its consequences in cardiovascular diseases.
Materials and Methods
Human umbilical vein endothelial cells (HUVEC) and Human dermal microvascular endothelial cells (HDMEC) were used for this study. Endothelial cells were treated in presence or absence of IL-6 (20ng/ml) for 36 hours and followed by insulin (100nM) stimulation for 15 minutes. Using immunoblotting, cell lysates were stained for phosphor- and total Akt levels to measure insulin resistance. To investigate changes in DNA methylation, cells were treated with or without neutrophil conditioned medium (NCM) as a physiological source of inflammation or IL-6 (at various concentrations) for 36 hours. Genomic DNA was processed for HPLC analysis for methyl cytosine content and cell lysates were analyzed for DNMT1 (DNA (cytosine-5)-methyltransferase 1) and DNMT3A (DNA (cytosine-5)-methyltransferase 3A) levels using immunoblotting.
Results
Endothelial cells stimulated with insulin exhibited an increase in phosphorylation of Aktser 473 in serum free conditions but such insulin response was not observed in cells treated with IL-6, suggesting chronic exposure of endothelial cells to IL-6 leads to insulin resistance. HPLC analysis for global DNA methylation resulted in decreased levels of 5-methyl cytosine in cells treated with pro-inflammatory molecules (both by NCM and IL-6) as compared to untreated controls. Subsequently, analysis in cells treated with IL-6 showed a significant decrease in DNMT1 levels but not in DNMT3A. Other pro-inflammatory marker such as TNF-α did not exhibit such changes.
Conclusion
Our study suggests: a) Chronic treatment of endothelial cells with IL-6 results in insulin resistance b) Neutrophil conditioned medium and IL-6 decreases methyl cytosine levels c) DNMT1 but not DNMT3a levels are reduced in cells treated with IL-6.
Sudarslal S, Ph.D.
Associate Professor, School of Biotechnology, Amrita University
Electrospray ionization ion trap mass spectrometry for cyclic peptide characterization
There has been considerable interest in the isolation and structural characterization of bioactive peptides produced by bacteria and fungi. Most of the peptides are cyclic depsipeptides characterized by the presence of lactone linkages and β-hydroxy fatty acids. Occurrence of microheterogeneity is another remarkable property of these peptides. Even if tandem mass spectrometers are good analytical tools to structurally characterize peptides and proteins, sequence analysis of cyclic peptides is often ambiguous due to the random ring opening of the peptides and subsequent generation of a set of linear precursor ions with the same m/z. Here we report combined use of chemical derivatization and multistage fragmentation capability of ion trap mass spectrometers to determine primary sequences of a series of closely related cyclic peptides.
![Delegate Talk: Bioanalytical Characterization of Therapeutic Proteins @ Amriteshwari Hall | Vallikavu | Kerala | India](http://www.amritabioquest.org/conference/2013/wp-content/uploads/sites/2/2015/02/bioanalt.jpg)
Ravindra Gudihal, Suresh Babu C V
Bioanalytical Characterization of Therapeutic Proteins
The characterization of therapeutic proteins such as monoclonal antibody (mAb) during different stages of manufacturing is crucial for timely and successful product release. Regulatory agencies require a variety of analytical technologies for comprehensive and efficient protein analysis. Electrophoresis-based techniques and liquid chromatography (LC) either standalone or coupled to mass spectrometry (MS) are at the forefront for the in-depth analysis of protein purity, isoforms, stability, aggregation, posttranslational modifications, PEGylation, etc. In this presentation, a combination of various chromatographic and electrophoretic techniques such as liquid-phase isoelectric focusing, microfluidic and capillary-based electrophoresis (CE), liquid chromatography (LC) and combinations of those with mass spectrometry techniques will be discussed. We present a workflow based approach to the analysis of therapeutic proteins. In successive steps critical parameters like purity, accurate mass, aggregation, peptide sequence, glycopeptide and glycan analysis are analyzed. In brief, the workflow involved proteolytic digestion of therapeutic protein for peptide mapping, N-Glycanase and chemical labeling reaction for glycan analysis, liquid-phase isoelectric focusing for enrichment of charge variants followed by a very detailed analysis using state of the art methods such as CE-MS and LC-MS. For the analysis of glycans, we use combinations of CE-MS and LC-MS to highlight the sweet spots of these techniques. CE-MS is found to be more useful in analysis of highly sialylated glycans (charged glycans) while nano LC-MS seems to be better adapted for analysis of neutral glycans. These two techniques can be used to get complementary data to profile all the glycans present in a given protein. In addition, microfluidic electrophoresis was used as a QC tool in initial screening for product purity, analysis of papain digestion fragments of mAb, protein PEGylation products, etc. The described workflow involves multiple platforms, provides an end to end solution for comprehensive protein characterization and aims at reducing the total product development time.