Freeze concentration for enrichment of nutrients in yellow water from no-mix toilets.

Separately collected urine ("yellow water") can be utilized as fertilizer. In order to decrease storage volumes and energy consumption for yellow water transport to fields, enrichment of nutrients in yellow water has to be considered. Laboratory-scale batch freeze concentration of yellow water has been tested in ice-front freezing apparatus: a stirred vessel and a falling film freeze concentrator (coolant temperatures: -6 to -16 degrees C). With progressing enrichment of the liquid concentrate, the frozen ice was increasingly contaminated with yellow water constituents (ammonia, total nitrogen, total phosphorus, TOC, and salts determined as conductivity). The higher the initial salinity of the yellow water and the lower the mechanical agitation of the liquid phase contacting the growing ice front, the more the frozen ice was contaminated. The results indicate, that in ice-front freezing devices multistage processes are necessary, i.e. the melted ice phase has to be purified (and the concentrates must be further enriched) in a second or even in a third stage. Energy consumption of this process is very high. However, technical scale suspension freeze concentration is reasonable in centralized ecological sanitation schemes if the population exceeds 0.5 million and distance of yellow water transportation to fields is more than 80 km.

Resonance Raman spectroscopy of sensory rhodopsin II from Natronobacterium pharaonis.

Sensory rhodopsin II (pSRII), the photophobic receptor from Natronobacterium pharaonis, has been studied by time-resolved resonance Raman (RR) spectroscopy using the rotating cell technique. Upon excitation with low laser power, the RR spectra largely reflect the parent state pSRII(500) whereas an increase of the laser power leads to a substantial accumulation of long-lived intermediates contributing to the RR spectra. All RR spectra could consistently be analysed in terms of four component spectra which were assigned to the parent state pSRII(500) and the long-lived intermediates M(400), N(485) and O(535) based on the correlation between the C = C stretching frequency and the absorption maximum. The parent state and the intermediates N(485) and O(535) exhibit a protonated Schiff base. The C = N stretching frequencies and the H/D isotopic shifts indicate strong hydrogen bonding interactions of the Schiff base in pSRII(500) and O(535) whereas these interactions are most likely very weak in N(485).

Sensory rhodopsin II from the haloalkaliphilic natronobacterium pharaonis: light-activated proton transfer reactions.

In the present work the light-activated proton transfer reactions of sensory rhodopsin II from Natronobacterium pharaonis (pSRII) and those of the channel-mutants D75N-pSRII and F86D-pSRII are investigated using flash photolysis and black lipid membrane (BLM) techniques. Whereas the photocycle of the F86D-pSRII mutant is quite similar to that of the wild-type protein, the photocycle of D75N-pSRII consists of only two intermediates. The addition of external proton donors such as azide, or in the case of F86D-pSRII, imidazole, accelerates the reprotonation of the Schiff base, but not the turnover. The electrical measurements prove that pSRII and F86D-pSRII can function as outwardly directed proton pumps, whereas the mutation in the extracellular channel (D75N-pSRII) leads to an inwardly directed transient current. The almost negligible size of the photostationary current is explained by the long-lasting photocycle of about a second. Although the M decay, but not the photocycle turnover, of pSRII and F86D-pSRII is accelerated by the addition of azide, the photostationary current is considerably increased. It is discussed that in a two-photon process a late intermediate (N- and/or O-like species) is photoconverted back to the original resting state; thereby the long photocycle is cut short, giving rise to the large increase of the photostationary current. The results presented in this work indicate that the function to generate ion gradients across membranes is a general property of archaeal rhodopsins.

The photophobic receptor from Natronobacterium pharaonis: temperature and pH dependencies of the photocycle of sensory rhodopsin II.

The photocycle of the photophobic receptor sensory rhodopsin II from N. pharaonis was analyzed by varying measuring wavelengths, temperature, and pH, and by exchanging H2O with D2O. The data can be satisfactorily modeled by eight exponents over the whole range of modified parameters. The kinetic data support a model similar to that of bacteriorhodopsin (BR) if a scheme of irreversible first-order reactions is assumed. Eight kinetically distinct protein states can then be identified. These states are formed from five spectrally distinct species. The chromophore states Si correspond in their spectral properties to those of the BR photocycle, namely pSRII510 (K), pSRII495 (L), pSRII400 (M), pSRII485 (N), and pSRII535 (O). In comparison to BR, pSRII400 is formed approximately 10 times faster than the M state; however, the back-reaction is almost 100 times slower. Comparison of the temperature dependence of the rate constants with those from the BR photocycle suggests that the differences are caused by changes of DeltaS. The rate constants of the pSRII photocycle are almost insensitive to the pH variation from 9.0 to 5.5, and show only a small H2O/D2O effect. This analysis supports the idea that the conformational dynamics of pSRII controls the kinetics of the photocycle of pSRII.

Heterologous coexpression of the blue light receptor psRII and its transducer pHtrII from Natronobacterium pharaonis in the Halobacterium salinarium strain Pho81/w restores negative phototaxis.

The photophobic receptor (psRII) and its transducer pHtrII from Natronobacterium pharaonis were heterologously coexpressed in the phototaxis-deficient Halobacterium salinarium strain Pho81/w which lacks all four bacterial rhodopsins, i.e. the two ion pumps bacteriorhodopsin and halorhodopsin as well as the two sensory pigments SRI and SRII. This genetically transformed Pho81/w strain showed a photophobic response upon illumination with blue light. The action spectrum of the psRII/pHtrII mediated phototactic behavior was determined in the range of 420-600 nm. The shape of the action spectrum was similar to the absorption spectrum of psRII, clearly indicating that the psRII-specific photophobic response in Pho81/w was restored. These results suggest that the pharaonis photoreceptor-transducer complex (psRII/pHtrII) is functionally competent to substitute the corresponding salinarium receptor system. Although the two archaea are phylogenetically quite distant from each other the two signal transduction chains are homologous systems which can replace each other.