Characteristics of continuous hydrogen production and fatty acid formation by an active hydrogen-producing anaerobic bacterium, Clostridium butyricum strain SC-E1, was examined under vacuum and non-vacuum culture systems. The continuous cultures were performed using 1040 ml anaerobic glass bottles containing 600 ml of medium including glucose and polypeptone at a concentration of 0.5 or 1.0% as substrate, and were conducted at pH 6.7, hydraulic retention time (HRT) 8h, and 30°C on a reciprocal shaker. The non-vacuum cultures at 16 days of incubation showed 2.0 to 2.3 mol-H2/mol-glucose and 1.4 to 2.0 mol-H2/mol-glucose of hydrogen productivity at 0.5 and 1.0% of substrate concentration, respectively. The vacuum cultures conducted at 0.28 atm gave 1.8 to 2.3 mol-H2/mol-glucose and 1.3 to 2.2 mol-H2/mol-glucose of hydrogen productivity at 0.5 and 1.0% of substrate concentration, respectively. The fatty acid production from the vacuum cultures exhibited approximately the same yield of fatty acids as those of the non-vacuum cultures. It was concluded that the maximal hydrogen production potential by anaerobic bacteria is 1.3 to 2.2 mol-H2/mol-glucose, which is less than 50% of theoretical. In addition, the total hydrogen production rate by a two-stage bioreactor consisting of a 1-litre anaerobic fermenter (HRT 10h) and a 4-litre photobioreactor (HRT 36h) feeding at 2.4-litre of 1.0% glucose per day was estimated at 1.4 to 5.6 mol-H2/mol-glucose, which is 12 to 47% theoretical.
Two cDNAs similar to aquaporins (AQPs) from other insect species were identified and characterized from the silkworm larva, Bombyx mori. The first cDNA (AQP-Bom1) cloned from the anterior silk gland encodes a 25 900 Da protein similar to insect AQPs isolated from several liquid-feeding insects. The second cDNA (AQP-Bom2) cloned from the posterior midgut encodes a 27 694 Da protein. Northern blot analysis has revealed that the AQP-Bom1 mRNA (2.3 kb) is expressed predominantly in the hindgut (colon and rectum), and moderately or minimally in the silk gland, midgut and Malpighian tubules, while the AQP-Bom2 mRNA (1.3 kb) is mainly expressed in the posterior midgut and Malpighian tubules. Functional analysis in Xenopus oocytes microinjected with the cRNA of these AQPs revealed that the AQP-Bom1 mRNA encodes a water-specific aquaporin, likely involved in the water retrieval function of the hindgut, while the AQP-Bom2 mRNA encodes an aquaglyceroporin, increasing glycerol and urea uptake.
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