Microbial Metabolism Systems Microbiology
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1 1 Microbial Metabolism Systems Microbiology Ching-Tsan Huang ( 黃慶璨 ) Office: Agronomy Hall, Room 111 Tel: (02) cthuang@ntu.edu.tw MIT OCW
2 Systems Microbiology aims to integrate basic biological information with genomics, transcriptomics, metabolomics, glycomics, proteomics and other data to create an integrated model of how a microbial cell or community functions. (Nature Review Microbiology) Systems Microbiology seeks to identify how microbial functions evolve and how emergent properties in cells and communities arise from seemingly simple, linear genetic sequences. 2
3 Source: U.S. Department of Energy Genome Programs 3
4 Source: U.S. Department of Energy Genome Programs 4
5 5 Why Systems Microbiology? Microbial systems are important and ideal models Microbes offer important lessons about all life forms
6 6
7 Disciplines Involving Systems Microbiology Microbiologists, Biochemists, Evolutionary biologists Mathematicians, Computer scientists, Physicists, Chemists, Control theorists, Systems engineers, Geochemists, Atmospheric chemists, Chemical and physical oceanographers, Earth scientists, and Biostatisticians. 7
8 8 Potential Applications of Systems Microbiology Agricultures Medicines Novel energy production systems Metabolic engineering Biocontrol Pollution and bioremediation Bioterrorism and decontamination Microbiological detection systems Global monitoring
9 9 Integrated Systems Microbiology Microbiology Multi-Disciplinary Microbial - Host System Genetics Chemistry Computer Science Biomedical Environmental Industrial Drug Discovery Antibiotic Resistance Diagnostics Vaccines Mathematics Biocontrol/ Bioremediation Bioreactors Bioenergy Biosensors Ecology Molecular Evolution
10 Utilization of Microbial Inoculants to Promote Sustainable Agriculture 10 Nutrients Signals Microbial Cell Factory Gene expression Primary metabolism Secondary Primary metabolism Secondary metabolism Phytostimulation Production of IAA ACC deaminase Increase rooting Increase Nitrogen uptake Reduced fertilizer usage Reduced nitrate pollution Bioremediation Clean-up of contaminated soils Biocontrol Antimicrobial Metabolites Reduced pathogens Reduced disease Reduced fungicide usage Biofertilization Nitrogen fixation Phosphate solubilization Reduced fertilizer usage Reduced water pollution
11 11 An Integrated Systems Biology Approach to Elucidating the Nature of Molecular Signalling in Microbial-Host Interactions Environmental signals Microbial signals Quorum sensing Overall Aim: Secondary metabolites Signal molecules Signalling and regulatory circuits Pseudomonas aeruginosa Cytokines Host signals Planktonic cells Biofilm formation To establish the importance of microbial and host signalling in determining the outcome of microbial-host interactions. Eukaryotic Host cells in culture
12 From Metagenomes to Ecosystem Functioning 12
13 Strain-resolved proteomics from environmental samples 13
14 From Proteins to Environments 14
15 From Proteins to Environments 15
16 16 Experimental Approaches in Systems Biology Sequencing technologies A capillary-based Sanger method on sequencing platforms. Massively higher throughput, such as 454 pyrosequencing, SOLiD and Solexa DNA microarrays and high-throughput sequencing Slides containing spatially organized DNA probes with known sequences for the identification and quantification of fluorescently labelled nucleic acids samples.
17 Next Generation Sequencing 17
18 18
19 19
20 Applications of Next Generation Sequencing 20 Category Complete genome resequencing Reduced representation sequencing Targeted genomic resequencing Paired end sequencing Metagenomic sequencing Small RNA sequencing Examples of applications Comprehensive polymorphism and mutation discovery in individual human genomes Large-scale polymorphism discovery Targeted polymorphism and mutation discovery Discovery of inherited and acquired structural variation Discovery of infectious and commensal flora microrna profiling
21 Applications of Next Generation Sequencing 21 Category Transcriptome sequencing Sequencing of bisulfitetreated DNA Chromatin immunoprecipitation sequencing (ChIP-Seq) Nuclease fragmentation and sequencing Molecular barcoding Examples of applications Quantification of gene expression and alternative splicing; transcript annotation; discovery of transcribed SNPs or somatic mutations Determining patterns of cytosine methylation in genomic DNA Genome-wide mapping of protein- DNA interactions Nucleosome positioning Multiplex sequencing of samples from multipleindividuals
22 22 Experimental Approaches in Systems Biology Proteomics technologies Mass spectrometry (MS)-based approaches for identification Array-based platforms Metabolomics technologies Powerful and accurately predictive approaches Bench-scale assays to high-throughput methods
23 23 Evolution of mass spectrometry-based proteomics from microbial isolates to communities
24 LC/MS-based Proteomics 24
25 Mass Spectrometry 25
26 Matrix-assisted laser desorption/ionization-based resequencing 26
27 27 Technical Challenges In Systems Microbiology Technical bottlenecks Single cell measurements Identifying and accurately measuring metabolites Cultivation Data accessibility Computational limitations Annotation and functional characterization of genes Proteomics High-throughput technologies
28 Tracking and segmenting single cells 28
29 29 Technical Challenges in Systems Microbiology Information and data gaps Acquiring information and data Assuring data quality Making systems data applicable and available A proposal for a systems microbiology database
30 30 Information and Data Gap Continued genome sequencing Data on the functions of the proteins or nucleic acids encoded by these sequences Accurate gene annotation Accurate quantitative information on a variety of biological parameters Data that indicate spatiotemporal relationships between system components
31 31 Information and Data Gap Data on community architecture Data that can reveal design principles and biological networks Data that expand the phenotypic characterizations of cells and communities Data on the natural variability of microbial systems Data on cell cycle or spatial events
32 32 Research in Systems Microbiology Regulation of biological systems Applying a systems biology approach to microbial communities Hypothesis-generating research Measuring noise in biological systems
33 Convergence of systems and synthetic biology 33
34 34 Reconstruction, validation and utilization of a metabolic reconstructi
35 Phases and data used to generate a metabolic reconstruction 35
36 Procedure to generate a biomass objective function 36
37 Network integration: the interface between different types of reconstruction 37
38 Reconstruction of transcriptional and translational networks 38
39 Timeline of events that resulted in the development of systems biology and synthetic biology 39
40 40
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