Dataset Information

Production of hydrogen from ?-1,4- and ?-1,4-linked saccharides by marine hyperthermophilic Archaea.

ABSTRACT: Nineteen hyperthermophilic heterotrophs from deep-sea hydrothermal vents, plus the control organism Pyrococcus furiosus, were examined for their ability to grow and produce H? on maltose, cellobiose, and peptides and for the presence of the genes encoding proteins that hydrolyze starch and cellulose. All of the strains grew on these disaccharides and peptides and converted maltose and peptides to H? even when elemental sulfur was present as a terminal electron acceptor. Half of the strains had at least one gene for an extracellular starch hydrolase, but only P. furiosus had a gene for an extracellular ?-1,4-endoglucanase. P. furiosus was serially adapted for growth on CF11 cellulose and H? production, which is the first reported instance of hyperthermophilic growth on cellulose, with a doubling time of 64 min. Cell-specific H? production rates were 29 fmol, 37 fmol, and 54 fmol of H? produced cell?¹ doubling?¹ on ?-1,4-linked sugars, ?-1,4-linked sugars, and peptides, respectively. The highest total community H? production rate came from growth on starch (2.6 mM H? produced h?¹). Hyperthermophilic heterotrophs may serve as an important alternate source of H? for hydrogenotrophic microorganisms in low-H? hydrothermal environments, and some are candidates for H? bioenergy production in bioreactors.

SUBMITTER: Oslowski DM

PROVIDER: S-EPMC3126455 | biostudies-literature | 2011 May

REPOSITORIES: biostudies-literature

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Production of hydrogen from α-1,4- and β-1,4-linked saccharides by marine hyperthermophilic Archaea.

Oslowski Daniel M DM Jung Jong-Hyun JH Seo Dong-Ho DH Park Cheon-Seok CS Holden James F JF

Applied and environmental microbiology 20110318 10

Nineteen hyperthermophilic heterotrophs from deep-sea hydrothermal vents, plus the control organism Pyrococcus furiosus, were examined for their ability to grow and produce H₂ on maltose, cellobiose, and peptides and for the presence of the genes encoding proteins that hydrolyze starch and cellulose. All of the strains grew on these disaccharides and peptides and converted maltose and peptides to H₂ even when elemental sulfur was present as a terminal electron acceptor. Half of the strains had a ...[more]

PMID: 21421788

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Project description:Thermococcus kodakarensis is a hyperthermophilic archaeon that harbors a complete set of genes for chitin degradation to fructose 6-phosphate. However, wild-type T. kodakarensis KOD1 does not display growth on chitin. In this study, we developed a T. kodakarensis strain that can grow on chitin via genetic and adaptive engineering. First, a chitinase overproduction strain (KC01) was constructed by replacing the chitinase gene promoter with a strong promoter from the cell surface glycoprotein gene, resulting in increased degradation of swollen chitin and accumulation of N-,N'-diacetylchitobiose in the medium. To enhance N-,N'-diacetylchitobiose assimilation in KC01, genes encoding diacetylchitobiose deacetylase, exo-?-d-glucosaminidase, and glucosamine-6-phosphate deaminase were also overexpressed to obtain strain KC04. To strengthen the glycolytic flux of KC04, the gene encoding Tgr (transcriptional repressor of glycolytic genes) was disrupted to obtain strain KC04?t. In both KC04 and KC04?t strains, degradation of swollen chitin was further enhanced. In the culture broth of these strains, the accumulation of glucosamine was observed. KC04?t was repeatedly inoculated in a swollen-chitin-containing medium for 13 cultures. This adaptive engineering strategy resulted in the isolation of a strain (KC04?tM1) that showed almost complete degradation of 0.4% (wt/vol) swollen chitin after 90 h. The strain produced high levels of acetate and ammonium in the culture medium, and, moreover, molecular hydrogen was generated. This strongly suggests that strain KC04?tM1 has acquired the ability to convert chitin to fructose 6-phosphate via deacetylation and deamination and further convert fructose 6-phosphate to acetate via glycolysis coupled to hydrogen generation.IMPORTANCE Chitin is a linear homopolymer of ?-1,4-linked N-acetylglucosamine and is the second most abundant biomass next to cellulose. Compared to the wealth of research focused on the microbial degradation and conversion of cellulose, studies addressing microbial chitin utilization are still limited. In this study, using the hyperthermophilic archaeon Thermococcus kodakarensis as a host, we have constructed a strain that displays chitin-dependent hydrogen generation. The apparent hydrogen yield per unit of sugar consumed was slightly higher with swollen chitin than with starch. As gene manipulation in T. kodakarensis is relatively simple, the strain constructed in this study can also be used as a parent strain for the development and expansion of chitin-dependent biorefinery, in addition to its capacity to produce hydrogen.

Dataset Information

Production of hydrogen from ?-1,4- and ?-1,4-linked saccharides by marine hyperthermophilic Archaea.

Publications

Production of hydrogen from α-1,4- and β-1,4-linked saccharides by marine hyperthermophilic Archaea.

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