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Prof. Jason K. Sello Department of Chemistry Brown University j ason_sello@brown

Towards Sustainable Living: Using Streptomyces Bacteria to Produce Renewable Energy and Commodity Chemicals from Plant Biomass. Prof. Jason K. Sello Department of Chemistry Brown University j ason_sello@brown.edu. Sources of Renewable Energy. WIND. SOLAR. BIOMASS. GEOTHERMAL. HYDRO.

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Prof. Jason K. Sello Department of Chemistry Brown University j ason_sello@brown

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  1. Towards Sustainable Living: Using Streptomyces Bacteria to Produce Renewable Energy and Commodity Chemicals from Plant Biomass Prof. Jason K. Sello Department of Chemistry Brown University jason_sello@brown.edu

  2. Sources of Renewable Energy WIND SOLAR BIOMASS GEOTHERMAL HYDRO

  3. Increasing World Biofuels Production • 15.9 billion gallons of biofuels were produced domestically in 2010 • 13.2 billion gallons of ethanol • 2.7 billion gallons of biodiesel • 138.6 billion gallons of gasoline was consumed in the US during 2010 BP Statistical Review of Energy June 2011. bp.com/statisticalreview

  4. Biotechnology for Conversion of Plant Biomass to Biofuels Rubin E.Genomics of cellulosic biofuels. Nature 454: 841-845, 2008.

  5. Plant BiomassFeedstocks Energy Crops (switch grass) Organic Trash Agricultural Residue Forestry Waste

  6. Biotechnology for Conversion of Plant Biomass to Biofuels Rubin E.Genomics of cellulosic biofuels. Nature 454: 841-845, 2008.

  7. Structural Components of Plant Biomass Lignin Hemicellulose Cellulose Rubin E.Genomics of cellulosic biofuels. Nature 454: 841-845, 2008.

  8. Structural Components of Plant Biomass Lignin Hemicellulose Cellulose Rubin E.Genomics of cellulosic biofuels. Nature 454: 841-845, 2008.

  9. Using Microorganisms for Biofuel Production Fermentation of yeast on plant sugars is currently used to produce bioethanol Engineered bacteria are being developed for the production of biodiesel by fermentation of plant sugars (Steen, Nature, 2010) Image by MarcinZemla and Manfred Auer, JBEI. http://newscenter.lbl.gov

  10. Synthetic Biology in Production of Biofuels Keasling and co-workers have engineered E. coli to convert hemicellulose into biofuels. Steen. Nature463, 559-564, 2010.

  11. Structural Components of Plant Biomass Lignin Hemicellulose Cellulose Rubin E.Genomics of cellulosic biofuels. Nature 454: 841-845, 2008.

  12. Lignin Component of Plant Biomass • Lignin constitute up to 30% of plant biomass • Highly stable and heterogeneous polymer consisting of aromatic building blocks • Lignin interferes with utilization of cellulose for the production of biofuels • Lignin can be enzymaticallydepolymerized by some bacteria and fungi • Bugg TD, Ahmad M, Hardiman EM & R Singh. Current Opinion in Biotechnology. 22:394–400, 2011.

  13. Phanerochaetechrysosporium diark.org P. chryosporium(white rot fungus) can consume lignin.

  14. Lignin Depolymerization • Bugg TD, Ahmad M, Hardiman EM & R Singh. Current Opinion in Biotechnology. 22:394–400, 2011.

  15. What is the fate of depolymerized lignin?

  16. Catabolism of Depolymerized Lignin (e.g., Sphingomonas) Masai E, Katayama Y, Fukuda M. Biosci. Biotechnol. Biochem., 71(1) 1-15, 2007.

  17. Catabolism of Depolymerized Lignin (e.g., Sphingomonas) Masai E, Katayama Y, Fukuda M. Biosci. Biotechnol. Biochem., 71(1) 1-15, 2007.

  18. Commodity Chemicals from TCA Cycle Triglycerides K. N. Timmis (ed.), Handbook of Hydrocarbon and Lipid Microbiology, 2010

  19. Biodiesel Alkyl ester R is methyl, ethyl, or propyl.

  20. Conversion of Triglycerides into Biodiesel 01.- 0.5% Sodium or Potassium Hydroxide Or Sodium Methoxide 80° Celsius Biodiesel (Fatty Acid Methyl Ester) Methanol Glycerin (Glycerol) Triglyceride (Triacylglycerols) Chemical reaction is a “trans-esterification”.

  21. Bioconversion of Lignin to Biofuels Triacyglycerols And Fatty Acids Acetyl-CoA Aromatic Compounds Lignin Succinyl-CoA

  22. An organism that can convert all the components of plant biomass into biofuels would be an efficient “biorefinery”.

  23. Complete Conversion of Lignocellulose to Biofuels Cellulose Triacyglycerols And Fatty Acids Acetyl-CoA Aromatic Compounds Lignin Succinyl-CoA Hemicellulose

  24. Prospecting for Plant Biomass Degraders

  25. “An antibiotic is a chemical substance produced by microbes that inhibits the growth of or even destroys other microbes” Selman Waksman (1888-1973)

  26. Timeline of Antibiotic Discovery

  27. Antibiotics in use as Anti-Bacterial Agents

  28. Antibiotics in use as Anti-Tumor Agents

  29. Antibiotics in Use as Immunosuppresants

  30. Diverse Morphologies and Colors of Streptomyces Species Image courtesy of T. Kieser

  31. Two Evolutionary Oddities Duckbill platypus Streptomycetes

  32. Streptomyces: An Unconventional Genus of Bacteria Multi-cellular Hyphalmorphology and mode of growth like fungi Complex life cycle Linear chromosomes and plasmids >8 Mb chromosomes are common Ubiquitous in terrestrial environments, easily cultured More than 500 species described Non-pathogenic relative of Mycobacterium tuberculosis Prodigious producers of antibiotics

  33. The Majority of Antibiotics are Produced by Streptomycetes

  34. Waksman screened soil samples in search of microorganisms that produce antibiotics. How can we identify microorganisms that degrade plant biomass?

  35. Identification of LigninolyticStreptomyces Strains S. lividans S. griseus S. coelicolor S. avermitilis S. natalensis S. chattanoogensis S. setonii S. badius S. viridosporus LigininolyticStreptomyces species can decolorize the aromatic dye, Azure B.

  36. Streptomyces viridosporus S. viridosporusis a bona fideligninolyticstreptomycete. It also is capable of consuming cellulose and hemicellulose. D.L. Crawford, Appl. Environ. Microbiol, 53: 2754-2760, 1987 D.L. Crawford, Appl. Environ. Microbiol, 41: 442-448, 1981 R L. Crawford, Appl. Environ. Microbiol, 45: 898-904, 1983

  37. Metagenomic-based Enzyme Discovery in LignocellulolyticMicrobial Communities DeAngelis, A. Bioengineering Research, 3, 146-158 (2010)

  38. Biodiversity in Tropical Forest Soil from Puerto Rico Richness (Number of Taxa DeAngelis,A. Bioeng. Res., 3, 146-158 (2010)

  39. Biodiversity in Tropical Forest Soil from Puerto Rico Richness (Number of Taxa DeAngelis,A. Bioeng. Res., 3, 146-158 (2010)

  40. Biodiversity in Lignin-Enriched Compost Richness (Number of Taxa Compost + Alkali Lignin Compost DeAngelis,A. Bioeng. Res., 3, 146-158 (2010)

  41. Biodiversity in Lignin-Enriched Compost Richness (Number of Taxa Compost + Alkali Lignin Compost

  42. Actinobacteria are Populous Soil Bacteria Mahidul University- Osaka University • Large group of terrestrial bacteria with high G+C content • genomes (e.g., Streptomyces, Corynebacteria, Nocardia, • Actinoplanes, and Mycobacteria). • - Many are filamentous like fungi • - Play a critical role in the decomposition of organic matter in soil • - Important organisms in biotechnology source of enzymes • and medicinal antibiotics

  43. Actinobacteria Produce Two-Thirds of the 23,000 Known Antibiotics Streptomyces derived compounds in red boxes

  44. Sir David A. Hopwood

  45. trpRS1 v Overview of Research in the Sello Group Chemical Synthesis and Drug Discovery Renewable Energy Streptomyces Bacteria cmlR Biosynthesis and Metabolomics Chemical Ecology Antibacterial Drug Resistance

  46. trpRS1 v Overview of Research in the Sello Group Chemical Synthesis and Drug Discovery Okandeji, JOC, 2008 Okandeji, JOC, 2009 Socha, BMC, 2010 Okandeji, BMC, 2011 Carney, JOC, 2012 Compton, ACS Chem. Biol. 2013 Nelson, mBio. 2013 Carney, JACS, 2014 Renewable Energy Socha, Energy & Fuels, 2010 Socha, OBC, 2010 Davis, AMB, 2010 Davis, J. Bacteriol., 2012 Davis, NAR, 2013 Davis, Genome Ann. 2013 Streptomyces Bacteria cmlR Antibacterial Drug Resistance Vecchione, J. Bacteriol., 2008 Vecchione, AAC, 2009 Vecchione, AAC, 2009 Vecchione, J. Bacteriol., 2010 Biosynthesis and Metabolomics Sello, J. Bacteriol., 2008 Badu-Nkansah, FEMS Lett., 2010 Totaro, ChemBioChem, 2012 Chemical Ecology Davis, Org. Lett., 2009 Morin, Org. Lett., 2010 Morin, OBC, 2012

  47. Actinobacteriaare Potential “Lignocellulose Biorefineries” • Gram-positive soil-dwelling bacteria • Degrade all components of plant biomass • Cellulose • Hemicellulose • Lignin • Naturally accumulate triacylglycerols, the precursors of biodiesel, and make commodity chemicals • Long history in industrial-scale fermentation for the production of antibiotics E. Wellington

  48. Plant Biomass-Degrading Actinobacteria Streptomyces viridosporus Amycolatopsissetonii A. setoniiand S. viridosporus are bona fideligninolytic bacteria. They also consume cellulose and hemicellulose. D.L. Crawford, Appl. Environ. Microbiol, 53: 2754-2760, 1987 D.L. Crawford, Appl. Environ. Microbiol, 41: 442-448, 1981 R L. Crawford, Appl. Environ. Microbiol, 45: 898-904, 1983

  49. The first bacterial lignin peroxidase was isolated from Streptomyces viridosporus Ramachandranet al. Appl. Environ. Microbiol. 53(12): 2754-2760, 1987.

  50. Lignin Depolymerization • Bugg TD, Ahmad M, Hardiman EM & R Singh. Current Opinion in Biotechnology. 22:394–400, 2011.

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