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Goal: Develop bacterial biocatalysts for consolidated bioprocessing (CBP) of lignocellulosis

Engineering Bacillus subtilis biocatalysts for production of biofuels and chemical feedstocks and biochemicals for pharmaceutical and nutraceutical applications. Mun Su Rhee, Lusha Wei, James F. Preston Department of Microbiology and Cell Science, University of Florida .

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Goal: Develop bacterial biocatalysts for consolidated bioprocessing (CBP) of lignocellulosis

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  1. Engineering Bacillus subtilis biocatalysts for production of biofuels and chemical feedstocks and biochemicals for pharmaceutical and nutraceutical applications. Mun Su Rhee, Lusha Wei, James F. PrestonDepartment of Microbiology and Cell Science, University of Florida Collaborators: L. O. Ingram, K. T. Shanmugam, W. Vermerris, J. Erickson, Support: FESC, USDA Biomass Research & Development Initiative Goal: Develop bacterial biocatalysts for consolidated bioprocessing (CBP) of lignocellulosis Secretion of cellulases and xylanases to process cellulose and xylans Fermentation of glucose and xylose for production of biofuelsand chemicals Objective: Engineer Bacillus subtilisfor CBP of methylglucurono(arabino)xylans (MeG(A)Xn) Engineer B. subtilis to convert MeGAXn to lactate for production of bioplastics Engineer B. subtilisto convert MeGXn to xylooligosaccharides : acidic (U-XOS) for production of anti-inflamatory agents and neutral (XOS) for production of prebiotics

  2. Interaction of Major Polymeric Sugars in Lignocellulosic Biomass Monocots: grasses, e.g. sorghum, sugarcane Methylglucuronoarabinoxylan, MeGAXn • Cellulose fibers form through hydrogen bonding interactions between individual cellulose stands • The noncarbohydrate polymer lignin embeds the interacting cellulose and glucuronoxylan through ester linkages to glucuronoxylan MeGAXn 18-35% MeGXn Cellulose fiber 32-48% Dicots: hardwoods, e.g.eucalyptus, sweetgum Methylglucuronoxylan, MeGXn

  3. Pretreatment of O-acetyl-glucuronoarabinoxylanand enzymatic depolymerization of 4-O-methylglucuronoxylan

  4. Schematic for bioconversion of MeGXn to U-XOS by B. subtilis strains.MeGX3, MeGX4or MeGX4-18 accumulate in the cultures of B.s. 168, MR42(ΔxynC) or MR44(ΔxynA), allowing production of XOS and/or U-XOS MeGXn (Sweetgumxylan) XynA (GH11) XynC (GH30) X2, X3 XOS MeGX4 MeGX4-12 MR44 (168, ΔxynA) MR42 (168, ΔxynC) XynA (GH11) XynC (GH30) XOS X2,X3 MeGX3 MeGX3 168

  5. Dicot : MeGXn Monocot : MeGAXn A, XOS

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