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We obtained efficient conversion of xylose to xylitol by transforming Saccharomyces cerevisiae with the gene encoding the xylose reductase (XR) of Pichia stipitis CBS 6054. Comparison of the chromosomal and cDNA copies of the XYL1 gene showed that the genomic XYL1 contains no introns, and an XR monomer of 318 amino acids (35,985 D) is encoded by an open reading frame of 954 bp. The amino acid sequence of the P. stipitis XR is similar to several aldose reductases, suggesting that P. stipitis XR is part of the aldoketo reductase superfamily. S. cerevisiae transformed with the XYL1 gene gave over 95% conversion of xylose into xylitol, a yield not obtainable with natural xylose utilizing yeasts.

The gradual shift of transportation fuels from oil based fuels to the alternative fuel resources and worldwide demand for energy has been the impetus for research to produce alcohol biofuels from renewable resources. Current bioethanol and biodiesel can, however, not cover an increasing demand for biofuels. Hence, there is an extensive need for advanced biofuels with superior fuel properties. The present review is focused on the developments of biobutanol, which is regarded to be superior to bioethanol in terms of energy density and hygroscopicity. Although acetone-butanolethanol (ABE) fermentation is one of the oldest large-scale fermentation processes, butanol yield by anaerobic fermentation remains sub-optimal. For sustainable industrial scale butanol production, a number of obstacles need to be addressed including choice of feedstock, low product yield, product toxicity to production strain, multiple end-products and downstream processing of alcohol mixtures.Metabolic engineering provides a means for fermentation improvements. Different strategies are employed in the metabolic engineering of Clostridia that aim to enhance the solvent production, improve selectivity for butanol production, and increase the tolerance of Clostridia to solvents. The introduction and expression of a non-clostridial butanol producing pathway in E. coli is most promising strategy for butanol biosynthesis. Several rigorous kinetic and physiological models for fermentation have been formulated, which form useful tool for optimization of the process. Due to the lower butanol titers in the fermentation broth, simultaneous fermentation and product removal techniques have been developed to improve production economics. With the use of new strains, inexpensive substrates, and superior reactor designs, economic ABE fermentation may further attract an attention of researchers all over the world. The present review is attempting to provide an overall outlook on discoveries and strategies that are being developed for commercial n-butanol production.

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This report describes the combined use of an enzyme-based glucose release system (EnBase®) and high-aeration shake flask (Ultra Yield Flask™). The benefit of this combination is demonstrated by over 100-fold improvement in the active yield of recombinant alcohol dehydrogenase expressed in E. coli. Compared to Terrific Broth and ZYM-5052 autoinduction medium, the EnBase system improved yield mainly through increased productivity per cell. Four-fold increase in oxygen transfer by the Ultra Yield Flask contributed to higher cell density with EnBase but not with the other tested media, and consequently the product yield per ml of EnBase culture was further improved.

Thermophilic Thermopolyspora flexuosa GH10 xylanase (TfXYN10A) was studied in the presence of biomass-dissolving hydrophilic ionic liquids (ILs) [EMIM]OAc, [EMIM]DMP and [DBNH]OAc. The temperature optimum of TfXYN10A with insoluble xylan in the pulp was at 65-70 °C, with solubilised 1 % xylan at 70-75 °C and with 3 % xylan at 75-80 °C. Therefore, the amount of soluble substrate affects the enzyme activity at high temperatures. The experiments with ILs were done with 1 % substrate. TfXYN10A can partially hydrolyse soluble xylan even in the presence of 40 % (v/v) ILs. Although ILs decrease the apparent temperature optimum, a surprising finding was that at the inactivating temperatures (80-90 °C), especially [EMIM]OAc increases the stability of TfXYN10A indicating that the binding of IL molecules strengthens the protein structure. Earlier kinetic studies showed an increased K with ILs, indicating that ILs function as competitive inhibitors. TfXYN10A showed low increase of K, which was 2-, 3- and 4-fold with 15 % [EMIM]OAc, [DBNH]OAc and [EMIM]DMP, respectively. One reason for the low competitive inhibition could be the high affinity to the substrate (low K ). Xylanases with low K (~1 mg/mL) appear to show higher tolerance to ILs than xylanases with higher K (~2 mg/mL). Capillary electrophoresis showed that TfXYN10A hydrolyses xylan to the end-products in 15-35 % ILs practically as completely as without IL, also indicating good binding of the short substrate molecules by TfXYN10A despite of major apparent IL binding sites above the catalytic residues. Substrate binding interactions in the active site appear to explain the high tolerance of TfXYN10A to ILs.