Purpose
A closed system that includes human metabolism Existing systems Apollo and Shuttle carried bottled oxygen and lithium-hydroxide
fine for a few days supplies too great to last months
Mir and ISS use electrolysis of water, recycled from dehumidifier and urine collection, CO2 removed with reusable sorbent
only half of oxygen breathed is recycled other half of oxygen breathed is sequestered in CO2 and dumped
Carbohydrate model
Carbohydrates are relatively simple to model. Starch, pectin, and dextrose are polysaccharides (C6H10O5)nC6H12O6 Human metabolism starts by hydrating to break into monosaccharide: (C6H10O5)nC6H12O6 + n H2O (n+1) C6H12O6 We can use formuli for monosaccharides to analyze all carbohydrates. Cellular respiration of monosaccharides is: 6 O2 + C6H12O6 6 H2O + 6 CO2
Replicating photosynthesis
Photosynthesis in plants occurs in an chloroplasts. This is a two step process: 1. Light reaction (Photophosporylation)
Capture light with chlorophyll Convert ADP into ATP, and NADP+ into NADPH Water is broken up, and oxygen released
2.
Chloroplast structure
highly structured biochemical machine Light reaction on surface of thylakoid Dark reaction in stroma and intermembrane space
Chlorophyl act as antennae to collect light pheophytin cleaves 2 H2O into O2, 4 H+, 4 ecytochrome complex pumps 4 more H+ into thylakoid interior NADP reductase converts NADP+, H+, and 2 e- into NADPH ATP synthase converts ADP and Pi into ATP, releases H+ from thylakoid
ATP Synthesis
H+ to ATP ratio 8:3 F0 portion is a stepper-motor F1 has 3 binding sites: produces 3 ATP per rotation Chloroplast F0 has 8 c-subunits:
releases 8 H+ ions per rotation
CO2 is added to RuBP That is broken into two molecules of 3PG ATP and NADPH used to attach phosphates and H to 3PG to create G3P From 12 molecules of G3P, two are removed to make glucose. The other ten are converted by ATP to reform 6 RuBP molecules.
Calvin-Benson cycle
The Calvin-Benson cycle can be summarized as:
6 CO2 + 18 ATP + 12 H2O + 12 NADPH + 12 H+ C6H12O6 + 18 Pi + 18 ADP + 12 NADP+
3 Pi 3 FDP 3 G3P 3 DHAP 3 F6P G6P Glucose 12 ADP 12 Pi 12 NADP+ 12 G3P 2 G3P 2 F6P 2 G3P Pi
12 ATP 12 NADPH 12 H+
2 DHAP
2 X5P 2 E4P
12 3PG
2 G3P
2 E4P 2 DHAP
6 CO2
6 ATP
6 Ru5P
2 R5P 2 X5P
Key: 3PG = 3-phosphogycerate G3P = glyceraldehydes 3-phosphate DHAP = dihydroxyacetone phosphate FDP = fructose 1,6-diphosphate F6P = fructose 6-phosphate G6P = glucose 6-phosphate E4P = erythrose 4-phosphate X5P = xylulose 5-phosphate SDP = sedoheptulose 1,7-diphosphate S7P = sedoheptulose 7-phosphate R5P = ribose 5-phosphate Ru5P = ribulose 5-phosphate RuBP = ribulose 1,5-biphosphate
4 X5P
Harvesting Chloroplasts
Grow pea seedlings in compost for 7-10 days at 18-22C
Light intensity should be relatively low (40-50 E/m2/s) Only young tissue (2-3 days after leaf emergence) should be used
Grid medium (0.35 M sucrose, 25 mM Hepes-NaOH, pH 7.6, 2mM EDTA) Sorbitol medium (50 mM Hepes-KOH, pH 8.4, 0.33 M sorbitol) 40% Percoll in sorbitol buffer 80% aqueous acetone Harvest leaves from pea seedlings and mix with semi-frozen grinding medium at a ratio of 20 g leaves per 100 ml medium. Homogenize the leaves with two 3 sec bursts of the polytron at 75% full speed. Strain the homogenate gently through eight layers of muslin to remove debris. Pour the suspension into 50 ml or 100 ml centrifuge tubes and centrifuge at 4000 g for 1 min. Discard the supernat in one motion (the pellets are quite firm at this stage) and wipe the inside of tubes. Resuspend the pellet gently in a small volume (4-8 ml) of sorbitol medium using a cotton swab or small paint brush, and layer the suspension on to an equal volume of 40% Percoll (Pharmacia) in sorbitol buffer. Centrifuge at 2500 g for 7 min (with the brake off). Intact chloroplasts are pelleted whereas lysed organelles fail to penetrate through the Percoll pad. Wash the pellet in 5 ml sorbitol medium and resuspend the pellet in 1 ml sorbitol medium. Check the intactness of the organelles under phase-contrast microscopy; intact organelles appear bright green, often with a surrounding halo, whereas broken chloroplasts appear darker and more opaque. The majority of the organelles (up to 95%) should be intact.
Capacity
2 light quanta to move one electron from H2O to NADP+ For each O2 molecule, 4 electrons, so 8 light quanta To evolve six molecules of O2, 48 light quanta must be absorbed 1 mole of light quanta = 72kcal @ 400nm, or 41kcal @ 700nm 6 moles of O2 requires 2496 kcal @ 52kcal / mole of light quanta 2496 kcal = 2.8981 kWh Humans require 0.84kg O2 per day, O2 masses 31.9988g per mole 12.68 kWh of light per person per day 1 mole of a subunit of polysaccharide (C6H10O5) masses 162.142g 4.256kg of carbohydrate produced per person per day
Equipment
Air filter Cabin air CO2 Sorbent Blower Return to cabin Compressor CO2 Storage
Chloroplast bag
Water pump
Starch outflow
References
U.S. Department of Energy, Federal Technology Alerts, http://www.pnl.gov/fta/13_glazings/13_glazings.htm Estrella Mountain Community College, http://www.emc.maricopa.edu/faculty/farabee/BIOBK/BioBookPS.html Photosynthesis: The Role of Light, http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/L/LightReactions.ht ml Lecture 10, ATP synthase, University of Illinois, http://www.life.uiuc.edu/crofts/bioph354/lect10.html Principles of Biochemistry, Albert L. Lehninger, Worth Publishers Inc. ISBN: 0-87901-136-X Plant Cell Biology, Harris and Oparka, 1994 Isolation of membranes and organelles from plant cells, Hall and Moore, 1983 A Nuclear-encoded RNA Polymerase in Corn Chloroplasts, Rachel Howard Yeast Extracts: Production, Properties and Components, 9th International Symposium on Yeasts, Sydney, August 1996, Rolf Sommer, Deutsche Hefewerke GmbH & Co. oHG