Fermentation of D-xylose to Ethanol by Saccharomyces cerevisiae CAT-1 Recombinant Strains.

Ethanol production by the D-xylose fermentation of lignocellulosic biomass would augment environmental sustainability by increasing the yield of biofuel obtained per cultivated area. A set of recombinant strains derived from the industrial strain Saccharomyces cerevisiae CAT-1 was developed for this purpose. First, two recombinant strains were obtained by the chromosomal insertion of genes involved in the assimilation and transport of D-xylose (Gal2-N376F). Strain CAT-1-XRT was developed with heterologous genes for D-xylose metabolism from the oxo-reductive pathway of Scheffersomyces stipitis (XYL1-K270R, XYL2); and strain CAT-1-XIT, with D-xylose isomerase (xylA gene, XI) from Streptomyces coelicolor. Moreover, both recombinant strains contained extra copies of homologous genes for xylulose kinase (XK) and transaldolase (TAL1). Furthermore, plasmid (pRS42K::XI) was constructed with xylA from Piromyces sp. transferred to CAT-1, CAT-1-XRT, and CAT-1-XIT, followed by an evolution protocol. After 10 subcultures, CAT-1-XIT (pRS42K::XI) consumed 74% of D-xylose, producing 12.6 g/L ethanol (0.31 g ethanol/g D-xylose). The results of this study show that CAT-1-XIT (pRS42K::XI) is a promising recombinant strain for the efficient utilization of D-xylose to produce ethanol from lignocellulosic materials.

Evaluation of sweet potato for fuel bioethanol production: hydrolysis and fermentation.

The enzymatic starch hydrolysis and bioethanol production from a variety of sweet potato developed for bioenergy purposes (K 9807.1) on the basis of its high starch yields, was studied. Drying at 55°C and 95°C of sweet potato neither affected the sugar content nor the starch enzymatic hydrolysis efficiency. Simultaneous saccharification and ethanol fermentations for dry matter ratio of sweet potato to water from 1:8 to 1:2 (w/v) were studied. Fresh sweet potato and dried at 55°C (flour) were assayed. At ratios of 1:8, similar results for fresh sweet potato and flour in terms of ethanol concentration (38-45 g/L), fermentation time (16 h) and sugar conversion (~ 100%) were found. At higher dry matter content, faster full conversion were observed using flour. A higher ratio than that for fresh sweet potato (1:2.2) did not improve the final ethanol concentration (100 g/L) and yields. High ethanol yields were found for VHG (very high gravity) conditions. The sweet potato used is an attractive raw matter for fuel ethanol, since up to 4800 L ethanol per hectare can be obtained.

Energy consumption evaluation of fuel bioethanol production from sweet potato.

The energy consumption for different operative conditions and configurations of the bioethanol production industrial process from an experimental variety of sweet potato (Ipomea batatas) K 9807.1 was evaluated. A process simulation model was developed using SuperPro Designer® software. The model was based on experimental data gathered from our laboratory experiments and technology and equipment suppliers. The effects of the dry matter ratio of sweet potato to water, the fermentation efficiency, and sweet potato sugar content, on the energy consumption (steam and electricity) were respectively evaluated. All factors were significant. The best ratio of dry matter to total water to work with fresh sweet potato was 0.2 kg dry sweet potato/kg water, as for greater ratios was not found a significant reduction in energy consumption. Also, the drying of the sweet potato previous its processing was studied. It presented an energy consumption greater than the energetic content of the bioethanol produced.

Extracellular enzymes produced by microorganisms isolated from maritime Antarctica.

Antarctic environments can sustain a great diversity of well-adapted microorganisms known as psychrophiles or psychrotrophs. The potential of these microorganisms as a resource of enzymes able to maintain their activity and stability at low temperature for technological applications has stimulated interest in exploration and isolation of microbes from this extreme environment. Enzymes produced by these organisms have a considerable potential for technological applications because they are known to have higher enzymatic activities at lower temperatures than their mesophilic and thermophilic counterparts. A total of 518 Antarctic microorganisms, were isolated during Antarctic expeditions organized by the Instituto Antártico Uruguayo. Samples of particules suspended in air, ice, sea and freshwater, soil, sediment, bird and marine animal faeces, dead animals, algae, plants, rocks and microbial mats were collected from different sites in maritime Antarctica. We report enzymatic activities present in 161 microorganisms (120 bacteria, 31 yeasts and 10 filamentous fungi) isolated from these locations. Enzymatic performance was evaluated at 4 and 20°C. Most of yeasts and bacteria grew better at 20°C than at 4°C, however the opposite was observed with the fungi. Amylase, lipase and protease activities were frequently found in bacterial strains. Yeasts and fungal isolates typically exhibited lipase, celullase and gelatinase activities. Bacterial isolates with highest enzymatic activities were identified by 16S rDNA sequence analysis as Pseudomonas spp., Psychrobacter sp., Arthrobacter spp., Bacillus sp. and Carnobacterium sp. Yeasts and fungal strains, with multiple enzymatic activities, belonged to Cryptococcus victoriae, Trichosporon pullulans and Geomyces pannorum.