Increasing cell biomass in Saccharomyces cerevisiae increases recombinant protein yield:the use of a respiratory strain as a microbial cell factory

BACKGROUND: Recombinant protein production is universally employed as a solution to obtain the milligram to gram quantities of a given protein required for applications as diverse as structural genomics and biopharmaceutical manufacture. Yeast is a well-established recombinant host cell for these purposes. In this study we wanted to investigate whether our respiratory Saccharomyces cerevisiae strain, TM6*, could be used to enhance the productivity of recombinant proteins over that obtained from corresponding wild type, respiro-fermentative strains when cultured under the same laboratory conditions. RESULTS: Here we demonstrate at least a doubling in productivity over wild-type strains for three recombinant membrane proteins and one recombinant soluble protein produced in TM6* cells. In all cases, this was attributed to the improved biomass properties of the strain. The yield profile across the growth curve was also more stable than in a wild-type strain, and was not further improved by lowering culture temperatures. This has the added benefit that improved yields can be attained rapidly at the yeast's optimal growth conditions. Importantly, improved productivity could not be reproduced in wild-type strains by culturing them under glucose fed-batch conditions: despite having achieved very similar biomass yields to those achieved by TM6* cultures, the total volumetric yields were not concomitantly increased. Furthermore, the productivity of TM6* was unaffected by growing cultures in the presence of ethanol. These findings support the unique properties of TM6* as a microbial cell factory. CONCLUSIONS: The accumulation of biomass in yeast cell factories is not necessarily correlated with a proportional increase in the functional yield of the recombinant protein being produced. The respiratory S. cerevisiae strain reported here is therefore a useful addition to the matrix of production hosts currently available as its improved biomass properties do lead to increased volumetric yields without the need to resort to complex control or cultivation schemes. This is anticipated to be of particular value in the production of challenging targets such as membrane proteins.

Simultaneous measurement of strain and temperature using the first- and second-order diffraction wavelengths of Bragg gratings

A method of discriminating between temperature and strain effects in fibre sensing using a conventionally written, in-fibre Bragg grating is presented. The technique uses wavelength information from the first and second diffraction orders of the grating element to determine the wavelength dependent strain and temperature coefficients, from which independent temperature and strain measurements can be made. The authors present results that validate this matrix inversion technique and quantify the strain and temperature errors which can arise for a given uncertainty in the measurement of the reflected wavelength.

Simultaneous measurement of strain and temperature using the first- and second-order diffraction wavelengths of Bragg gratings

A method of discriminating between temperature and strain effects in fibre sensing using a conventionally written, in-fibre Bragg grating is presented. The technique uses wavelength information from the first and second diffraction orders of the grating element to determine the wavelength dependent strain and temperature coefficients, from which independent temperature and strain measurements can be made. The authors present results that validate this matrix inversion technique and quantify the strain and temperature errors which can arise for a given uncertainty in the measurement of the reflected wavelength.

Bragg grating in polymer optical fibre for strain, bend and temperature sensing

We report on a Bragg grating written in an eccentric-cored polymer optical fibre for measurement of strain, bend and temperature. The strain sensitivity achieves 1.13 pm µe -1. The temperature response shows a negative sign with the thermal sensitivity of -50.1 pm ?C-1. For bend sensing, this device exhibits a strong fibre orientational dependence, wide bend curvature range of ±22.7 m-1 and a high bend sensitivity of 63.3 pm/m-1.

22km distributed strain sensor using Brillouin loss in an optical fibre

A novel distributed strain sensor is presented utilizing the strain dependence of the frequency at which the Brillouin loss is maximized in the interaction between a cw laser and a pulsed laser. A strain resolution of 20 µ with a spatial resolution of 5 m has been achieved with a 22 km sensing length.

Simultaneous measurement of temperature and strain: cross sensitivity considerations

Interferometric sensors using optical fibers as a transduction medium have been shown to be sensitive to a variety of physical measurands. A result of this is that the resolution of a system designed to sense strain, for example, may be compromised by fluctuations in the temperature of the environment. The possibility of simultaneously determining the strain and temperature applied to the same piece of highly birefringent fiber is discussed. Second-order effects are shown to be important for long sensing lengths or in the presence of high strains or temperature changes. The results of experiments carried out to verify the theoretical predictions are also described.

The mechanics of creep crack growth in a Magnox alloy

Magnox AL80 has been used for a study of creep crack propagation. A number of variables have been considered such as specimen geometry,notch root radius, material thickness, creep prestrain and stress level.The work has covered the material behaving under two values of the creep exponent, n=3.5 and n=7, according to the stress level. As well as observing initiation times and crack growth rates, scribed grids have been used to examine the near crack tip strain levels and distributions. It was shown that estimations of COD from notch flank opening can give misleading indications of material behaviour and that a more informative method was to monitor displacements in the material surrounding the crack tip. Strong evidence was found for crack advance being displacement controlled, however it was shown that the COD approach should be considered geometry dependant. The summation of ∈xx and ∈yy provided the most successful description of crack advance as it produced a single value that described propagation in all the cases concidered. The strain distributions indicates that σyy was related to distance from a point ahead of the crack tip by the exponent - (l/n+l) and that σxx is proportional to σyy. The constraint stresses arising in the DEN and CN specimens were evaluated. Initiation time was found to be principally affected by the stress level but was modified by the constraints arising from specimen geometry. Crack growth was found not to obey either the empirical K or σpett relationships but was reviewed in context of the observed strain behaviour.

Fibre Bragg grating strain sensors

The fabrication of in-fibre Bragg gratings (FBGs) and their application as sensors is reported. The strain and temperature characteristic results for a number of chirped and uniform gratings written into three different host fibres are presented. The static and dynamic temperature response of a commercially available temperature compensated grating is reported. A five sensor wavelength division multiplexed fibre Bragg grating strain measurement system with an interrogation rate of 25 Hz and resolution of 10 was constructed. The results from this system are presented. A novel chirped FBG interrogation method was implemented in both the 1.3 and 1.5 m telecommunication windows. Several single and dual strain sensor systems, employing this method, were constructed and the results obtained from each are reported and discussed. These systems are particularly suitable for the measurement of large strain. The results from a system measuring up to 12 m and with a potential measurement range of 30 m are reported. This technique is also shown to give an obtainable resolution of 20 over a measurement range of 5 000 for a dual sensor system. These systems are simple, robust, passive and easy to implement. They offer low cost, high speed and, in the case of multiple sensors, truly simultaneous interrogation. These advantages make this technique ideal for strain sensing in SMART structures. Systems based on this method have been installed in the masts of four superyachts. A system, based on this technique, is currently being developed for the measurement of acoustic waves in carbon composite panels. The results from an alternative method for interrogating uniform FBG sensors are also discussed. Interrogation of the gratings was facilitated by a specifically written asymmetric grating which had a 15 nm long linearly sloped spectral edge. This technique was employed to interrogate a single sensor over a measurement range of 6 m and two sensors over a range of 4.5 me. The results obtained indicated achievable resolutions of 47 and 38 respectively.

Photonic skin for pressure and strain sensing

In this paper, we report on the strain and pressure testing of highly flexible skins embedded with Bragg grating sensors recorded in either silica or polymer optical fibre. The photonic skins, with a size of 10cm x 10cm and thickness of 1mm, were fabricated by embedding the polymer fibre or silica fibre containing Bragg gratings in Sylgard 184 from Dow Corning. Pressure sensing was studied using a cylindrical metal post placed on an array of points across the skin. The polymer fibre grating exhibits approximately 10 times the pressure sensitivity of the silica fibre and responds to the post even when it is placed a few centimetres away from the sensing fibre. Although the intrinsic strain sensitivities of gratings in the two fibre types are very similar, when embedded in the skin the polymer grating displayed a strain sensitivity approximately 45 times greater than the silica device, which also suffered from considerable hysteresis. The polymer grating displayed a near linear response over wavelength shifts of 9nm for 1% strain. The difference in behaviour we attribute to the much greater Young's modulus of the silica fibre (70 GPa) compared to the polymer fibre (3 GPa).

22km distributed strain sensor using Brillouin loss in an optical fibre

A novel distributed strain sensor is presented utilizing the strain dependence of the frequency at which the Brillouin loss is maximized in the interaction between a cw laser and a pulsed laser. A strain resolution of 20 µ with a spatial resolution of 5 m has been achieved with a 22 km sensing length.

Bend sensing by measuring the resonance splitting of long-period fiber gratings

An application of long-period fiber gratings (fabricated in standard fiber) as bend sensors is reported. A simple model, taking account of the strain and compression in the fiber cladding, is used to calculate the splitting of the cladding modes and is found to be in excellent agreement with the experimental results. Over 80 nm resonance splitting was observed under a 5.6 m-1 bend curvature giving a bend sensitivity of 14.5 nm/(m-1), the best obtained data so far. © 2001 Elsevier Science B.V.

Strain and temperature sensitivity of a singlemode polymer optical fibre

We report experimental measurements of the strain and temperature sensitivity of the optical phase in a singlemode polymer optical fibre. These values were obtained by measuring optical path length change using a Mach-Zender interferometer.

Controlling the properties of microstructured plastic optical fiber Bragg gratings using acousto-optic excitation

A fine control of the mPOF Bragg grating spectrum properties, such as maximum reflected power and 3dB bandwidth, through acousto-optic modulation (AOM) using flexural regime is presented. A numerical comparison of the strain field along mPOFBG - AOM and the similar structure with SMFBG-AOM was presented, showing that the strain field amplitude is higher along the mPOFBG due to its smaller mechanical stiffness. The obtained results can be used in the development of fine-tuned optical filters using low voltage sources and low frequency regimes, to obtain tunable optical filters and to control the shape of the spectrum. Studies of the behavior in different gratings (such as phase shifted and long period gratings) for photonic applications, such as tunable notch filters or tunable cavities, are in progress. It can potentially be applied on tunable optical filters for POF transmission. © 2012 IEEE.

Annealing effects on strain and stress sensitivity of polymer optical fibre based sensors

The annealing effects on strain and stress sensitivity of polymer optical fibre Bragg grating sensors after their photo-inscription are investigated. PMMA optical fibre based Bragg grating sensors are first photo-inscribed and then they were placed into hot water for annealing. Strain, stress and force sensitivity measurements are taken before and after annealing. Parameters such as annealing time and annealing temperature are investigated. The change of the fibre diameter due to water absorption and the annealing process is also considered. The results show that annealing the polymer optical fibre tends to increase the strain, stress and force sensitivity of the photo-inscribed sensor.