Rewiring cellular metabolism for heterologous biosynthesis of Taxol

Abstract

Taxol is one of the anticancer drugs synthesized naturally in the evergreen Taxus brevifolia forest tree belonging to the yew family (Taxaceae) growing on the Pacific. There are reportedly evidence for treating ovarian, breast and lung cancers through this drug given its unique structural and functional features. Extraction of this drug from yew trees bark is one of the most common ways of producing this drug, but 3000 trees are needed to obtain a kilogram of Taxol. Hence, further attention has recently been attracted to the metabolic engineering strategies, including, engineering cellular metabolism of microorganisms and their optimization. Accordingly, the present paper article was aimed to review recent advances in elevating the production and commercialization of Taxol through metabolic engineering techniques.     

Optimal Control for Switched Systems with Pre-defined Order and Switch-Dependent Dynamics

The optimal control problem for switched systems, with a predefined order of switches, is considered. The differential equations may depend explicitly on previous switching instants, and the latter may be state dependent. The solution is based on the Calculus-of-Variations, which leads to a single two-point boundary-value problem. A new condition for the Hamiltonian jump at the switching instants is obtained. Simple numerical examples demonstrate the results.     

Spin Selectivity in Electron Transfer in Photosystem I

Photosystem I (PSI) is one of the most studied electron transfer (ET) systems in nature; it is found in plants, algae, and bacteria. The effect of the system structure and its electronic properties on the electron transfer rate and yield was investigated for years in details. In this work we show that not only those system properties affect the ET efficiency, but also the electrons’ spin. Using a newly developed spintronic device and a technique which enables control over the orientation of the PSI monolayer relative to the device (silver) surface, it was possible to evaluate the degree and direction of the spin polarization in ET in PSI. We find high‐spin selectivity throughout the entire ET path and establish that the spins of the electrons being transferred are aligned parallel to their momenta. The spin selectivity peaks at 300 K and vanishes at temperatures below about 150 K. A mechanism is suggested in which the chiral structure of the protein complex plays an important role in determining the high‐spin selectivity and its temperature dependence. Our observation of high light induced spin dependent ET in PSI introduces the possibility that spin may play an important role in ET in biology.     

The bimolecular hydrogen-deuterium exchange behavior of protonated alkyl dipeptides in the gas phase

As part of an ongoing characterization of the intrinsic chemical properties of peptides, thermal hydrogen-deuterium exchange has been studied for a series of fast-atom-bombardment-generated protonated alkyldipeptides and related model compounds in the reaction with D₂O, CH₃OD, and ND₃ in a Fourier transform ion cyclotron resonance mass spectrometer. Despite the very large basicity difference between the dipeptides and the D₂O and CH₃OD exchange reagents, efficient exchange of all active hydrogen atoms occurs. From the kinetic data it appears that exchange of the amino, amide, and hydroxyl hydrogens proceeds with different efficiencies, which implies that the proton in thermal protonated dipeptides is immobile. The selectivity of the exchange at the different basic sites is governed by the nature of both the dipeptide and the exchange reagent. The results indicate that reversible proton transfer in the reaction complexes, which effectuates the deuterium incorporation, is assisted by formation of multiple hydrogen bonds between the reagents. Exchange is considered to proceed via the intermediacy of different competing intermediate complexes, each of which specifically leads to deuterium incorporation at different basic sites. The relative stabilization of the competing intermediate complexes can be related to the relative efficiencies of deuterium incorporation at different basic sites in the dipeptide. For all protonated dipeptides studied, the exchange in the reaction with ND₃ proceeds with unit efficiency, whereas all active hydrogen atoms are exchanged equally efficiently. Evidently specific multiple hydrogen bond formations are far less important in the reversible proton transfers with the relatively basic ammonia, which allows effective randomization of all active hydrogen atoms in the reaction complexes.     

Real-time analysis of partially polarized light with a space-variant subwavelength dielectric grating

A novel method for real-time polarization measurement is presented. The method is based on a space-variant wave plate that we realized as a computer-generated space-variant subwavelength dielectric grating. The Stokes parameters of the incident beam are determined by Fourier analysis of the space-variant intensity transmitted through the grating and an analyzer. We discuss the design and realization of such wave plates and demonstrate our technique with polarization measurements of both polarized and partially polarized CO(2)-laser radiation at a wavelength of 10.6 mum.     

Space-variant Pancharatnam-Berry phase optical elements with computer-generated subwavelength gratings

Space-variant Pancharatnam-Berry phase optical elements based on computer-generated subwavelength gratings are presented. By continuously controlling the local orientation and period of the grating we can achieve any desired phase element. We present a theoretical analysis and experimentally demonstrate a Pancharatnam-Berry phase-based diffraction grating for laser radiation at a wavelength of 10.6microm.     

Formation of helical beams by use of Pancharatnam-Berry phase optical elements

Spiral phase elements with topological charges based on space-variant Pancharatnam-Berry phase optical elements are presented. Such elements can be achieved by use of continuous computer-generated space-variant subwavelength dielectric gratings. We present a theoretical analysis and experimentally demonstrate spiral geometrical phases for infrared radiation at a wavelength of 10.6microm .     

Critical behavior of light in mode-locked lasers

Light is shown to exhibit critical and tricritical behavior in passively mode-locked lasers with externally injected pulses. It is a first and unique example of critical phenomena in a one-dimensional many-body light-mode system. The phase diagrams consist of regimes with continuous wave, driven parapulses, spontaneous pulses via mode condensation, and heterogeneous pulses, separated by phase transition lines that terminate with critical or tricritical points. Enhanced non-Gaussian fluctuations and collective dynamics are present at the critical and tricritical points, showing a mode system analog of the critical opalescence phenomenon. The critical exponents are calculated and shown to comply with the mean field theory, which is rigorous in the light system.     

Study of the membrane effect on turbulent mixing measurements in shock tubes

The effect of the thin membrane on the time evolution of the shock wave induced turbulent mixing between the two gases initially separated by it is investigated using two different sets of experiments. In the first set, in which a single-mode large-amplitude initial perturbation was studied, two gas combinations (air/SF and air/air) and two membrane thicknesses were used. The main conclusion of these experiments was that the tested membrane has a negligible effect on the evolution of the mixing zone, which evolves as predicted theoretically. In the second set, in which similar gas combinations and membrane thicknesses were used, small amplitude random-mode initial perturbation, caused by the membrane rupture, rather than the large amplitude single-mode initial perturbation used in the first set, was studied. The conclusions of these experiments were: (1) The membrane has a significant effect on the mixing zone during the initial stages of its growth. This has also been observed in the air/air experiment where theoretically no growth should exist. (2) The membrane effect on the late time evolution, where the mixing zone width has reached a relatively large-amplitude, was relatively small and in good agreement with full numerical simulations. The main conclusion from the present experiments is that the effect of the membrane is important only during the initial stages of the evolution (before the re-shock), when the perturbations have very small amplitudes, and is negligible when the perturbations reach relatively large amplitudes. Received 29 August 1998 / Accepted 25 December 1998