Anhydrobiosis in yeast: FT-IR spectroscopic studies of yeast grown under conditions of severe oxygen limitation.

Anhydrobiosis is a unique state of living organisms when metabolism is temporarily and reversibly delayed in response to the extreme desiccation of cells. The production of dry active preparations of yeast grown under anaerobic conditions is not currently possible because preparations are extremely sensitive to the dehydration procedure, though they could be very helpful in different biotechnological processes, including bioethanol production. To characterize mechanisms responsible for such sensitivity to the dehydration procedure, Fourier transform infrared spectroscopy was used to study the composition of aerobically grown yeast Saccharomyces cerevisiae resistant to dehydration and grown under conditions of severe oxygen limitation and sensitive to dehydration. Results indicated that significantly lower amounts of lipids in cells, grown under conditions of severe oxygen limitation, may be related to the mechanisms of sensitivity. Dehydration of both resistant and sensitive S. cerevisiae cells was accompanied by similar changes in main cellular compounds. Amounts of nucleic acids and proteins decreased slightly, whereas that of lipids and carbohydrates increased. Artificially reduced sensitivity to dehydration in S. cerevisiae cells, grown under conditions of severe oxygen limitation, led to the increase in the lipid concentration. The chemical composition of S. cerevisiae membranes is proposed to dictate the resistance to dehydration in resistant and sensitive cells.

Hydrogen-producing Escherichia coli strains overexpressing lactose permease: FT-IR analysis of the lactose-induced stress.

The lactose permease gene (lacY) was overexpressed in the septuple knockout mutant of Escherichia coli, previously engineered for hydrogen production from glucose. It was expected that raising the lactose transporter activity would elevate the intracellular lactose concentration, inactivate the lactose repressor, induce the lactose operon, and as a result stimulate overall lactose consumption and conversion. However, overexpression of the lactose transporter caused a considerable growth delay in the recombinant strain on lactose, resembling to some extent the "lactose killing" phenomenon. Therefore, the recombinant strain was subjected to selection on lactose-containing media. Selection on plates with 3% lactose yielded a strain with a decreased content of the recombinant plasmid but with an improved ability to grow and produce hydrogen on lactose. Macromolecular analysis of its biomass by means of Fourier transform-infrared spectroscopy demonstrated that increase of the cellular polysaccharide content might contribute to the adaptation of E. coli to lactose stress.

Lactobacillus acidophilus La5 and Bifidobacterium lactis Bb12 cell surface hydrophobicity and survival of the cells under adverse environmental conditions.

Changes in the cell surface hydrophobicity (CSH) of probiotic bacteria Lactobacillus acidophilus La5 and Bifidobacterium lactis Bb12 and the survival of these cells were examined in response to varied cultivation conditions and adverse environmental conditions. An inverse linear relationship (P < 0.01) was detected between the CSH of intact L. acidophilus La5 and B. lactis Bb12 and survival of cells subjected to subsequent freezing/thawing, long-term storage or exposure to mineral and bile acids. The observed relationships were supported by significant correlations between the CSH and changes in composition of the cell envelopes (proteins, lipids and carbohydrates) of L. acidophilus La5 and B. lactis Bb12 examined using FT-IR spectroscopy and conventional biochemical analysis methods. The results also suggest that the estimates of hydrophobicity, being a generalized characteristic of cell surfaces, are important parameters to predict the ability of intact probiotic bacteria to endure extreme environments and therefore should be monitored during cultivation. A defined balance of cell components, which can be characterized by the reduced CSH values, apparently helps to ensure the resistance, improved viability and hence the overall probiotic properties of bacteria.

Application of FT-IR spectroscopy for control of the medium composition during the biodegradation of nitro aromatic compounds.

Previous studies showed that cabbage leaf extract (CLE) added to the growth medium can noticeably promote the degradation of nitro aromatic compounds by specific consortium of bacteria upon their growth. For further development of the approach for contaminated soil remediation it was necessary to evaluate the qualitative and/or quantitative composition of different origin CLE and their relevance on the growth of explosives-degrading bacteria. Six CLE (different by species, cultivars and harvesting time) were tested and used as additives to the growth medium. It was shown that nitro aromatic compounds can be identified in the FT-IR absorption spectra by the characteristic band at 1,527 cm(-1), and in CLE by the characteristic band at 1,602 cm(-1). The intensity of the CLE band at 1,602 cm(-1) correlated with the concentration of total nitrogen (R2=0.87) and decreased upon the growth of bacteria. The content of nitrogen in CLE differed (0.22-1.00 vol.%) and significantly influenced the content of total carbohydrates (9.50-16.00% DW) and lipids [3.90-9.90% dry weight (DW)] accumulated in bacterial cells while the content of proteins was similar in all samples. Though this study showed quantitative differences in the composition of the studied CLE and the response of bacterial cells to the composition of the growth media, and proved the potential of this additive for remediation of contaminated soil. It was shown that analysis of CLE and monitoring of the conversion of nitro aromatic compounds can be investigated by FT-IR spectroscopy as well as by conventional chemical methods.