Photochemistry of a retinal protonated schiff-base analogue mimicking the opsin shift of bacteriorhodopsin

A retinal Schiff base analogue which artificially mimics the protein-induced red shifting of absorption in bacteriorhodopsin (BR) has been investigated with femtosecond multichannel pump probe spectroscopy. The objective is to determine if the catalysis of retinal internal conversion in the native protein BR, which absorbs at 570 nm, is directly correlated with the protein-induced Stokes shifting of this absorption band otherwise known as the "opsin shift". Results demonstrate that the red shift afforded in the model system does not hasten internal conversion relative to that taking place in a free retinal-protonated Schiff base (RPSB) in methanol solution, and stimulated emission takes place with biexponential kinetics and characteristic timescales of approximately 2 and 10.5 ps. This shows that interactions between the prosthetic group and the protein that lead to the opsin shift in BR are not directly involved in reducing the excited-state lifetime by nearly an order of magnitude. A sub-picosecond phase of spectral evolution, analogues of which are detected in photoexcited retinal proteins and RPSBs in solution, is observed after excitation anywhere within the intense visible absorption band. It consists of a large and discontinuous spectral shift in excited-state absorption and is assigned to electronic relaxation between excited states, a scenario which might also be relevant to those systems as well. Finally, a transient excess bleach component that tunes with the excitation wavelength is detected in the data and tentatively assigned to inhomogeneous broadening in the ground state absorption band. Possible sources of such inhomogeneity and its relevance to native RPSB photochemistry are discussed.     

Fragrance release profile from sonochemically prepared protein microsphere containers

Protein microspheres have been prepared by sonicating a mixture of pure fragrant oil (amyl acetate (AA)) with an aqueous protein (bovine serum albumin) solution. The prepared protein spheres are nano- to micrometer sized with an encapsulation efficiency of approx. 97% for the AA present on the surface and inside the BSA capsule. Containers were found stable for more than 6 months when stored sealed at 4 °C and 20 °C. For the release profile measurements, we used a simple, automated and direct method. We continuously weighed the encapsulated microspheres and measured the evaporation rates. The release profiles at 15 °C and 25 °C display two different evaporation rates. The higher rate is the sum of a few evaporation rates, including water molecules, while the slower rate is due to the evaporation of pure AA. The changes in the evaporation rates occur upon the collapse of the container. This event coincides with the full evaporation of water. For morphological characterization we dyed the AA with Nile red, and used SEM, ESEM, Cryo-SEM, light microscopy, and confocal laser scanning microscopy measurements.     

Tracking excited state dynamics with coherent control: automated limiting of population transfer in LDS750

Closed loop automated pulse shaping experiments are conducted to investigate population transfer in solutions of the laser dye LDS750 in acetonitrile and ethanol. Guided by a genetic algorithm, the optical phases of broadband noncollinear parametric amplifier pulses are modulated by a micromachined deformable mirror to minimize sample fluorescence. The objectives were to test if nonlinearly chirped pulses could reduce population transfer below levels attained by their linearly chirped analogues, and if so, whether the resulting pulse shapes could be rationalized in terms of the photoinduced molecular dynamics. We further aimed to discover how the optimal solutions depend on the pulse fluence, and on the nature of the solvent. Using frequency resolved optical gating, the optimal field is shown to consist of a transform limited blue portion, which promotes population to the excited state, and a negatively chirped red tail, which follows the Stokes shifting of the excited density and dumps it back down to the ground state through stimulated emission. This is verified by comparing the optimal group delay dispersion with multichannel transient absorption data collected in acetonitrile. The optimal pulse shape was not significantly affected by variation of pulse fluence or by the change of solvent for the two polar liquids investigated. These results are discussed in terms of accumulated insights concerning the photophysics of LDS750 and the capabilities of our learning feedback scheme for quantum control.     

A new spectral window on retinal protein photochemistry

A VIS pump/hyperspectral NIR probe study of all-trans-retinal protonated Schiff base (RPSB) in ethanol is presented. Upon irradiation, a short-lived absorption band covers the recorded range of λ = 1-2 μm. It decays to reveal the tail of S(1) emission at λ < 1.3 μm, along with a residual absorption at longer wavelengths, both of which decay with the known kinetics of internal conversion to S(0). The existence of this hitherto unrecorded excited-state absorption deep in the NIR will require a revision of current models for RPSB electronic structure. The phenomenological similarity of these observations with ultrafast NIR studies of carotenoids raises the question of whether three, and not two, electronic states participate in RPSB photochemistry as well. The relevance of these observations to retinal protein photochemistry is discussed.