PEGylation of membrane proteins like bacteriorhodopsin as a tool to increase their stability toward ethanol.

Protection of biological compounds, for example, enzymes, viruses, or even whole cells, against degradation is very important for many applications. Embedding of such compounds into polymer matrices is a straightforward common method. However, in biotechnology and medicine there is a great interest to prepare micro- and nanosized shells around the biocomponents in order to protect them and having only a minor increase in size. The PEGylation of biological macromolecules has gained attention because degradation by proteolytic enzymes is significantly retarded and, in turn, their bioavailability is enhanced. We found that PEGylation is also a powerful tool to protect biomaterials from degradation by small organic solvent molecules, in particular, ethanol. Methoxy-polyethylene glycol (MPEG) modified BR survives exposure to significant concentrations of ethanol, up to 30%, and preserves its photochromism, whereas unmodified PM is instantaneously denatured at such concentrations. This is useful for potential technical applications of BR but is of relevance for many other applications where biomaterials and, in particular, biomembranes may be exposed to solvents.

Photoreversibility and photostability in films of octopus rhodopsin isolated from octopus photoreceptor membranes.

In this work, a new biomaterial resulting from the isolation of octopus rhodopsin (OR) starting from octopus photoreceptor membranes is presented. Mass spectroscopic characterization was employed in order to verify the presence of rhodopsin in the extract. Photoreversibility and photochromic properties were investigated using spectrophotometric measurements and pulsed light. Thin films of OR were realized using the gel-matrix entrapment method in polyvinyl alcohol solution. The results indicate that the photoreversibility and the photostability of the OR in gel-matrices are maintained. Several measurements were performed to test the stability of the resulting biomaterial in time and at room temperature. Preliminary tests demonstrate that the photoreversibility and the photostability are still found after few days from the biomaterial preparation and after the exposure for several hours at room temperature.

Thermochromism of bacteriorhodopsin and its pH dependence.

Purple membranes (PMs), which consist of the photochromic membrane protein bacteriorhodopsin (BR) and lipids only, show complex thermochromic properties. Three different types of reversible temperature-dependent spectral transitions were found, involving spectral states absorbing at 460, 519, and 630 nm. These thermochromic absorption changes were analyzed in the range from 10 to 80 degrees C. In dependence on the bulk pH value, hypsochromic or bathochromic shifts in the BR absorption spectra are observed in BR gels as well as in BR films. The thermochromic changes between both purple and blue or purple and red were quantified in the CIE color system. The molecular changes causing these effects are discussed, and a model is presented in terms of intramolecular protonation equilibriums. The thermochromic properties of BR may be of interest in applications like security tags, as this feature may complement the well-known photochromic properties of BR.

Biomolecular optical data storage and data encryption.

The use of bacteriorhodopsin (BR) as an active layer in write-once-read-many optical storage is presented. This novel feature of BR materials may be used on a wide variety of substrates, among them transparent substrates but also paper and plastics. The physical basis of the recording process is polarization-sensitive two-photon absorption. As an example for this new BR application, an identification card equipped with an optical recording strip is presented, which has a capacity of about 1 MB of data. The recording density currently used is 125 kB/cm2, which is far from the optical limits but allows operation with cheap terminals using plastic optics. In the examples given, data are stored in blocks of 10 kB each. A special optical encryption procedure allows the stored data to be protected from unauthorized reading. The molecular basis of this property is again the polarization-sensitive recording mechanism. The unique combination of optical storage, photochromism, and traceability of the BR material is combined on the single-molecule level. BR introduces a new quality of storage capability for applications with increased security and anticounterfeiting requirements.