Patents and literature

Protein engineering and site-directed mutagenesis is becoming immensely important in both fundamental studies and commercial applications involving proteins and enzymes in biocatalysis. Protein engineering has become a powerful tool to help biochemists and molecular enzymologists elucidate structure-function relationships in enzymic active sites, to understand the intricacies of protein folding and denaturation, and to alter the selectivity of enzymatic catalysis. Commercial applications of engineered enzymes are being developed to increase protein stability, widen or narrow substrate specificity, and to develop novel approaches for use of enzymes in organic synthesis, drug design, and clinical applications. In addition to protein engineering, novel expression systems have been designed to prepare large quantities of genetically engineered proteins. Recent US patents and scientific literature on protein engineering, site-directed mutagenesis, and protein expression systems related to protein engineering are surveyed. Patent abstracts are summarized individually and a list of literature references are given.     

Zur Theorie der Chiralitätsfunktionen

The Algebraic Theory of Chirality Functions is derived by means of exclusively qualitative considerations. Hence, the significance of quantitative results is questionable. Moreover the construction of “Näherungsansätze” (”Approximation-Ansatz”), which may be interpreted as semiempirical methods, is achieved on the basis of plausibility and mathematical simplicity. Since physical arguments are not included, the consistency or inconsistency of “Näherungsansätze” with experimental results do not justify direct physical conclusions.     

The 6Li,6Li-INADEQUATE Experiment: A New Tool for 1D- and 2D-NMR Studies of Organolithium Compounds

The 1D- and 2D-⁶Li, ⁶Li-INADEQUATE experiments are described as new tools for the detection of scalar coupled nonequivalent ⁶Li nuclei in organolithium clusters. Practical applications of these sequences are demonstrated for the ⁶Li-NMR spectra of (E)-1-lithio-2-(2-lithiophenyl)-1-phenylhex-1-ene ( 1 ) and (E)-2-lithio-1-(2-lithiophenyl)-1-phenylpent-1-ene ( 2 ), where signals due to dimers and monomers can be distinguished. The performance of the 2D-⁶Li, ⁶Li-INADEQUATE and the ⁶Li, ⁶Li-COSY-45-LR experiment are compared. The ⁶Li chemical shifts of 1 and 2 are discussed.     

HPLC-NMR of fatty alcohol ethoxylates

The application of HPLC-NMR for the analysis of a mixture of fatty alcohol ethoxylates (FAEs) is described. The use of the new generation, cryogenically cooled probes is investigated in respect of the sensitivity advantage that they provide. The FAE mixture is separated using liquid chromatography at the critical point of adsorption. The ability of the method to differentiate between the different end groups and the degree of polymerization of the mixture constituents is investigated. Both on-flow and stop-flow HPLC-NMR techniques were used together with two-dimensional NMR spectroscopy. The results are compared with those obtained by using an evaporative light scattering detector for the HPLC.     

CHEMICALLY MODIFIED PHOTOSYNTHETIC BACTERIAL REACTION CENTERS: CIRCULAR DICHROISM, RAMAN RESONANCE, LOW TEMPERATURE ABSORPTION, FLUORESCENCE AND ODMR SPECTRA AND POLYPEPTIDE COMPOSITION OF BOROHYDRIDE TREATED REACTION CENTERS FROM Rhodobacter sphaeroides R26

Abstract— Reaction centers from Rhodobacter sphaeroides have been modified by treatment with sodium borohydride similar to the original procedure [Ditson et al., Biochim. Biophys. Acta 766 , 623 (1984)], and investigated spectroscopically and by gel electrophoresis.
(1) Low temperature (1.2 K) absorption, fluorescence, absorption- and fluorescence-detected ODMR, and microwave-induced singlet-triplet absorption difference spectra (MIA) suggest that the treatment produces a spectroscopically homogeneous preparation with one of the 'additional' bacteriochlorophylls being removed. The modification does not alter the zero field splitting parameters of the primary donor triplet (^TP870).
(2) From the circular dichroism and Raman resonance spectra in the1500–1800 cm^-1 region, the removed pigment is assigned to Bchl_M, e.g. the "extra" Bchl on the "inactive" M-branch.
(3) A strong coupling among all pigment molecules is deduced from the circular dichroism spectra, because pronounced band-shifts and/or intensity changes occur in the spectral components assigned to all pigments. This is supported by distinct differences among the MIA spectra of untreated and modified reaction centers, as well as by Raman resonance.
(4) The modification is accompanied by partial proteolytic cleavage of the M-subunit. The preparation is thus spectroscopically homogeneous, but biochemically heterogenous.     

Der chemische Transport von Platin mit Chlor. Experimentell bearbeitet mit Marita Trenkel

The Chemical Transport of Platinum with Chlorine Experiments show that the chemical transport of platinum by means of chlorine within a temperature gradient at temperatures below ≈ 1000°K goes into the hot temperature region, but at higher temperatures in the reverse direction. From the thermodynamic discussion it can be seen, that the platinum content of the gas phase at low temperatures is governed by the exothermic formation of Pt₆Cl_12,g, and at higher temperatures by the endothermic formation of PtCl_3,g and PtCl_2,g. The platinum content of the gas phase passes a minimum at ≈ 1000°K, if P(Cl₂) = 3.5 atm. This result is in agreement with the observed inversion of the transport direction.     

Massenspektrometrische Beobachtungen und chemische Transportexperimente mit den Systemen VCl3/Al2Cl6 und VCl2/Al2Cl6

Mass Spectroscopic Observations and Chemical Transport Experiments with the Systems VCl₃/Al₂Cl₆ and VCl₂/Al₂Cl₆ By mass spectrometry the equilibrium VCl_3,s + 0.5 Al₂Cl_6,g ? VAlCl_6,g has been determined: ΔH°(298) = 25.6(±0.5) kcal; ΔS°(298) = 23.0(±3) cal/K, ΔCp (assumed) = ?4 cal/K. This is approximately in agreement with results determined by ligand field spectroscopy by ANUNDSKÅS and ØYE (A. and Ø.). For the dimerization of VCl_3,g values for ΔH and ΔS have been derived. The molecule VAl₂Cl₉ assumed by A. and Ø. could not been observed by mass spectrometry. For the VCl₂/Al₂Cl₆ complex, observed by chemical transport, A. and Ø. give the formula VAl₃Cl₁₁. This molecule could not been observed by mass spectrometry. This suggests a smaller concentration, compared with the results of A. and Ø. Stabilization of VCl_2,s (by metal-nietal-bonds) shifts the reaction to the left, whith explains the lower complex concentration as well as the larger molecular weight of the complex. With chlorides stabilized by stronger metal-metal bonds (MoCl₃, MoCl₂, Nb₃Cl₈) AlCl₃ complexes are not formed in observable concentrations. The chemical transport of VCl₂ with Al₂Cl₆ needs relatively high temperatures (973 → 873 K). In this case the addition of SiCl₄ hinders the attack of the quartz ampoule by Al₂Cl₆. Using a VCl₃ + VCl₂ mixture, VCl₃ is transported by Al₂Cl₆ (673 → 623 K) into the colder region. If afterwords the ampoule is reversed, VCl₃ again moves into the colder region, but the thermal decomposition of VCl₃ at the same time causes that a VCl₂-residue remains in the hot region.