Genesis of the BioSolarH2 Team

AFOSR MURI June 2005 launch Renewable Bio-solar Hydrogen Production from Roust Oxygenic Phototrophs

Founding Members BioSolarH2Æ Team Charles Dismukes PU photosynthesis/inorganic Edward Stiefel PU enzymology/inorganic Donald Bryant PSU genetics/mol biology cyanos Matthew Posewitz CSM genetics/mol biology algae Eric Hegg MiSU enzymology/inorganic Robert Bidigare UH microbiology/ecology BioSolarH2Æ Team 2007 MURI supported team (other support)

•10 co-PIs LC-MS & NMR research staff: •3 senior researchers J. Eng •10 young postdoctoral coworkers I. Pelczer •14 doctoral candidates

2005 MURI 2006 AF Supplement 2007 Eastern subgroup Western subgroup AF

PU PSU PU Stiefel CSM UH MiSU PU MoSU AFRL GIT Dismukes Bryant Posewitz Bidigare Hegg Austin Peters Pachter Spain

Ananyev, Shen Smith Brown, Keymer Boyd, Trohalaki Kolling(ACS) Vogl (‘08) Drobnini, Messer Hatlevik Galajda McGlynn, Cheng (CSC, Beer (NBTC) Zadvorny, ’08) Ruebush Carrieri Garcia- Carrieri Meuser, Liao Mulder, Ranchou- McNeely Costas McNeely Nagy (DoD) McGlynn Peyruse Bennette Xu Lambert Skizim (DOE) Guerra (Fulbright) What makes the AFOSR BioSolarH2 MURI format successful? The Goal

Quantitative Microbial Tools & Models Ecology

Renewable Bio-solar H2 Production from Robust Oxygenic Phototrophs

Gene Expression Photosynthesis & Manipulation Evolution What makes the AFOSR BioSolarH2 MURI program successful? Motivation The secret to success is communication: triannual reports & teleconferences -BioSolarH2Æ Face-to-Face Annual Meetings

July 2005, Princeton July 2006, Princeton August 2007, Breckenridge

10 distinguished international advisors Our MURI team works together Internal Collaborations

Nomadic PIs and students eager to wander Posewitz Lab, Jon Meuser, Bryant Lab: Yu Xu, Bidigare Lab: Susan Brown, Dismukes Lab: Kelsey McNeely, Damian Carrieri, Peters Lab: Shawn McGlynn •14 publications, 2 in queue, 12 in preparation •4 specialty instruments built for this project •3 LC-MS instruments purchased for the price of one •PU central support for MS/NMR metabolomics & proteomics Educational impact of the BioSolarH2 MURI •10 young PhD coworkers •14 doctoral candidates in training Undergraduate Trainees PU PSU PU CSM UH MiSU PU MoSU GIT Dismukes Bryant Stiefel Posewitz Bidigare Hegg Austin Peters Spain

T. Brown Byrne ‘08 A. Chambers C.Muldoon K.Swanson C. Berne C. Louie W. Stephens C. Howe J. Fox J. Kouch, P. Curtin, Y. Lee, M. Honigberg , A Cortez, A. Ma, S. Kriston, B. Nitchke; L. Fogli, P. Wu,, A. Bautista, D. Chien, J. Chou, N. Slater

MURI spawned new or revised courses Stiefel, Freshman Seminar: Elements of Life Dismukes, GEO255 Astrobiology (Bioprospecting Lab) Dismukes, ENV525: Production of Renewable Fuels and Energy Leveraged $ Support & Synergistic Collaborations “Translational Activities” Austin Lab: Peters Lab: Nikolay Zorin, Pushchino 6.19% Bidigare Lab: Joan Broderick, MSU 7.13% Robert Szilagyi, MSU 9.98% 33.3% Bryant Lab: 4.75e-006% Posewitz Lab: 2.49% John Golbeck, PSU Bonnie Baxter Westminster

Dave Ward, MoSU Arthur Grossman, Stanford 5.77% Igor Brown, NASA Michael Seibert, NREL 2.38% Samples: O. Lenz, K. Kovacs, 4.28% Maria Ghirardi, NREL J. Appel 28.5% Dismukes Lab: Total = $22.6 Rabinowitz, PU Rabitz, PU AFOSR MURI Gajda Ajlani, CEA Saclay AFOSR core Ben Hankamer U. Queensland DOD-DURIP Olaf Kruse, U. Bielfeld, GDR NSF: Photosystems Igor Brown, NASA DOE-BES: Half-Cell Patricia Brletic W&J DOE-JGI: Genomes DOE-JGI: Metagenomes Hegg Lab: DOE-OS+BER+GTL: H2 Networks Jim Fredrickson at PNNL DOE-GTL: Metabolomics Helen Kreuzer-Martin at PNNL DOE-EERE: H2 Program Robert Hausinger MSU DOE-Bioenergy Center C. Peter Wolk MSU Kazem Kashefi MSU Gemma Reguera MSU Claire Vieille MSU Teresa Theil MSU Tim Donohue,UW Madison Daniel Noguera UW Madison Public Website (new site under construction)

Public Announcement for 2007 BioSolarH2 Mid-Term Review 2007 BioSolarH2 Registration Form

RENEWABLE BIO-SOLAR HYDROGEN PRODUCTION FROM ROBUST OXYGENIC PHOTOTROPHS Project Announcement FA9550-05-1-0365

BioSolar Team Milestones Research Presentations July 2006 2nd BioSolarH2 symposium Robert Bidigare - University of Hawaii Feb 1, 2006: BioSolarH2 team expands John Peters - Montana State University Dr. Steven Chu visits Princeton Lab Charles Dismukes - Princeton University July 2005 Goals and Payoff Pacific Rim Meeting Presentation Kick Off Meeting Program and Photos IGERT Meeting Presentation Renewable,Renewable, Clean,Clean, WaterWater--BasedBased

BiosolarBiosolar HH2 Green Algae & Cyanobacteria Yield Hydrogen, Lipids & Sugars Light Reactions PSI and PSII Dark Reactions

Triglyceride (Oil) Biodiesel light chlorophyll _ Lipid Synthesis 4 e CO 4 H+ 2 Carbohydrate Synthesis Ethanol 2 H2O water-splitting enzyme H -generating O2 2 H2 hydrogenase

Algae Cyanobacteria • Hydrogen, carbohydrate and lipids are made by both algae and cyanobacteria • Hydrogen can be made in light and dark • Algae are good at photo-production of H2 and cyanobacteria are good at dark-production of H2 2 um

20 um Renewable Bio-solar Hydrogen Production from Robust Oxygenic Phototrophs H2 AFOSR-MURI

mutants

scr ee The H2 “Z-scheme” ni ng field g en om e

g en es Oxygenic Photosynthesis

HoxE HoxF ORF? HoxU HoxY HoxH Nco I (3375) Nco I (6934) Bam HI (1484) Eco RI (4977) Nco I (1099) Cla I (2981) Cla I (7047)

diaphorase moiety Ni-Fe hydrogenase moiety O genomic sequence around hox genes in S. platensis H O 2 7098 bp 2 GM O OriginalNew PI Objectives Objectives (>2006) s •DirectedDevelop newevolution H2 sensors using for arrayed rapid, microchemostats sensitive, HTP screening & DNA shuffling •Isolation/characterizationScreen CMMED collection of & cyanobacterial field sites for right NAD-linked phototrophs hydrogenase •HydrogenaseExamine hypercarbonate-requiring catalytic mechanism via strain, computational Arthrospira chemistry maxima •Isolate and sequence hydrogenases genes from halophiles •Bioprospecting for symbiotic H2-exchanging microbes •Random & specific mutagenesis for higher H2 production •Genetically combining host phototrophs with optimal hydrogenases