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High-yield hydrogen production from biomass by in vitro metabolic engineering: Mixed sugars coutilization and kinetic modeling.

Proceedings of the National Academy of Sciences of the United States of America (2015-04-08)
Joseph A Rollin, Julia Martin del Campo, Suwan Myung, Fangfang Sun, Chun You, Allison Bakovic, Roberto Castro, Sanjeev K Chandrayan, Chang-Hao Wu, Michael W W Adams, Ryan S Senger, Y-H Percival Zhang
ABSTRACT

The use of hydrogen (H2) as a fuel offers enhanced energy conversion efficiency and tremendous potential to decrease greenhouse gas emissions, but producing it in a distributed, carbon-neutral, low-cost manner requires new technologies. Herein we demonstrate the complete conversion of glucose and xylose from plant biomass to H2 and CO2 based on an in vitro synthetic enzymatic pathway. Glucose and xylose were simultaneously converted to H2 with a yield of two H2 per carbon, the maximum possible yield. Parameters of a nonlinear kinetic model were fitted with experimental data using a genetic algorithm, and a global sensitivity analysis was used to identify the enzymes that have the greatest impact on reaction rate and yield. After optimizing enzyme loadings using this model, volumetric H2 productivity was increased 3-fold to 32 mmol H2⋅L(-1)⋅h(-1). The productivity was further enhanced to 54 mmol H2⋅L(-1)⋅h(-1) by increasing reaction temperature, substrate, and enzyme concentrations--an increase of 67-fold compared with the initial studies using this method. The production of hydrogen from locally produced biomass is a promising means to achieve global green energy production.

MATERIALS
Product Number
Brand
Product Description

Xylose, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Glycerol solution, puriss. p.a., 86-89% (T)
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Glycerol solution, puriss., meets analytical specification of Ph. Eur., BP, 84-88%
USP
Glycerin, United States Pharmacopeia (USP) Reference Standard
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Glycerin, meets USP testing specifications
Supelco
Glycerin, Pharmaceutical Secondary Standard; Certified Reference Material
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Glycerol solution, 83.5-89.5% (T)
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Glycerol, FCC, FG
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D-(+)-Xylose, ≥99%
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Glycerol, ReagentPlus®, ≥99.0% (GC)
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Glycerol, ≥99.5%
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Glycerol, BioXtra, ≥99% (GC)
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Glycerol, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for electrophoresis, ≥99% (GC)
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Glycerol, for molecular biology, ≥99.0%
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D-(+)-Xylose, ≥99%
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D-(+)-Xylose, BioXtra, ≥99%
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Glucose Isomerase from Streptomyces murinus, ≥350 U/g
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Glycerol, BioUltra, for molecular biology, anhydrous, ≥99.5% (GC)
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Glycerol, tested according to Ph. Eur., anhydrous
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D-(+)-Xylose, BioUltra, ≥99.0% (sum of enantiomers, HPLC)
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Glycerol, puriss. p.a., ACS reagent, anhydrous, dist., ≥99.5% (GC)
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Glycerol, analytical standard
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Glycerol, ACS reagent, ≥99.5%
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Glycerol, puriss., meets analytical specification of Ph. Eur., BP, USP, FCC, E422, anhydrous, 99.0-101.0% (alkalimetric)