Bioproduction of cis‐(1S,2R)‐indandiol,a chiral pharmaceutical intermediate,using a solid–liquid two‐phase partitioning bioreactor for enhanced removal of inhibitors

BACKGROUND: A solid‐liquid two‐phase partitioning bioreactor (TPPB) was used in the biotransformation of indene to cis‐(1S,2R)‐indandiol by Pseudomonas putida 421‐5 (ATCC 55687). Metered substrate feeding in single‐phase operation, or delivery from an immiscible liquid, have previously been employed to regulate the exposure of the biocatalyst to inhibitory concentrations of the substrate. In contrast, the solid‐liquid platform provided in situ substrate addition (ISSA) as well as simultaneous it in situ product removal (ISPR) as a means of overcoming substrate and product toxicity. Three different modes of operation were compared for their effects on the performance of this biotransformation: single‐phase, fed‐batch operation was carried out as a benchmark in 2.75 L aqueous medium, and subsequently with the inclusion of either 700 g liquid silicone oil or 700 g solid polymer beads. RESULTS: Biphasic modes achieved a 3‐fold productivity improvement with respect to single‐phase (30 to 90 mg L^?1 h^?1), and solid‐liquid productivity was similar to liquid‐liquid operation while achieving more extensive removal of inhibitory compounds resulting in a slightly higher product titer (1.29 vs 1.16 g L^?1). The operability of the reactor was improved by the phase stability of the solid polymer beads relative to immiscible organic solvents, preventing emulsion formation and facilitating analytics. CONCLUSION: Solid polymer beads replaced the immiscible liquid auxiliary phase for substrate delivery while performing simultaneous inhibitory molecule sequestration. Copyright © 2011 Society of Chemical Industry     

Optimization of Robust Loss Functions for Weakly-Labeled Image Taxonomies

The recently proposed ImageNet dataset consists of several million images, each annotated with a single object category. These annotations may be imperfect, in the sense that many images contain multiple objects belonging to the label vocabulary. In other words, we have a multi-label problem but the annotations include only a single label (which is not necessarily the most prominent). Such a setting motivates the use of a robust evaluation measure, which allows for a limited number of labels to be predicted and, so long as one of the predicted labels is correct, the overall prediction should be considered correct. This is indeed the type of evaluation measure used to assess algorithm performance in a recent competition on ImageNet data. Optimizing such types of performance measures presents several hurdles even with existing structured output learning methods. Indeed, many of the current state-of-the-art methods optimize the prediction of only a single output label, ignoring this ‘structure’ altogether. In this paper, we show how to directly optimize continuous surrogates of such performance measures using structured output learning techniques with latent variables. We use the output of existing binary classifiers as input features in a new learning stage which optimizes the structured loss corresponding to the robust performance measure. We present empirical evidence that this allows us to ‘boost’ the performance of binary classification on a variety of weakly-supervised labeling problems defined on image taxonomies.     

Spectroscopic and Theoretical Study on Electronically Modified Chromophores in LOV Domains: 8‐Bromo‐ and 8‐Trifluoromethyl‐Substituted Flavins

Two chemically synthesized flavin derivatives, 8‐trifluoromethyl‐ and 8‐bromoriboflavin (8‐CF₃RF and 8‐BrRF), were photochemically characterized in H₂O and studied spectroscopically after incorporation into the LOV domain of the blue light photoreceptor YtvA from Bacillus subtilis. The spectroscopic studies were paralleled by high‐level quantum chemical calculations. In solution, 8‐BrRF showed a remarkably high triplet quantum yield (0.97, parent compound riboflavin, RF: 0.6) and a small fluorescence quantum yield (0.07, RF: 0.27). For 8‐CF₃RF, the triplet yield was 0.12, and the fluorescence quantum yield was 0.7. The high triplet yield of 8‐BrRF is due to the bromine heavy atom effect causing a stronger spin–orbit coupling. Theoretical calculations reveal that the decreased triplet yield of 8‐CF₃RF is due to a smaller charge transfer and a less favorable energetic position of T₂, required for intersystem crossing from S₁ to T₁, as an effect of the electron‐withdrawing CF₃ group. The reconstitution of the LOV domain with the new flavins resulted in the typical LOV photochemistry, consisting of triplet state formation and covalent binding of the chromophore, followed by a thermal recovery of the parent state, albeit with different kinetics and photophysical properties.     

SUPERCRITICAL EXTRACTION OF LINSEED OIL: ECONOMICAL VIABILITY AND MODELING EXTRACTION CURVES

The aim of the present study was to use supercritical technology to recover linseed oil (Linum usitatissimum L.) using carbon dioxide (alone or modified with ethanol as solvent) to determine the influence of the technique on the chemical composition of the oil obtained, model the kinetic curves of extraction, and estimate the manufacturing cost of the process. The experiments were conducted at 323 K, pressure of 25 MPa, constant solvent flow of 1.7 × 10^?5 kg/s, and extraction time of 5 h. The highest yield was obtained with the addition of cosolvent (28.8%). The SFE process of linseed oil manufacture proved to be economically viable, resulting in a product with a specific cost of 13.21 US$/kg_oil. As to oil composition, the main fatty acids detected were linolenic and oleic acid.     

Microstructured Fischer‐Tropsch Reactor Scale‐up and Opportunities for Decentralized Application

Current projects focusing on the energy transition in traffic will rely on a high‐level technology mix for their commissioning. One of those technologies is the Fischer‐Tropsch synthesis (FTS) that converts synthesis gas into hydrocarbons of different chain lengths. A microstructured packed‐bed reactor for low‐temperature FTS is tested towards its versatility for biomass‐based syngas with a high inert gas dilution. Investigations include overall productivity, conversion, and product selectivity. A 60‐times larger pilot‐scale reactor is further tested. Evaporation cooling is introduced which allows to increase the available energy extraction from the system. From that scale on, an autothermal operation at elevated conversion levels is applicable.     

Cry Woolf

The Woolf Reforms, recently instituted to make civil litigation more user friendly, targeted, amongst other things, the investigator’s best friend, the Anton Piller Order (AP). The Order was targeted directly, by having its name changed to the Search Order, and indirectly, in that Woolf aimed to make claimants expend more effort in finding solutions before resorting to the “nuclear option” of the AP.     

Selection of catalytically active biotin ligase and trypsin mutants by phage display

Phage display has been shown to facilitate greatly the selectionof polypeptides with desired properties by establishing a directlink between the polypeptide and the gene that encodes it. However,selection for catalytic activities displayed on phage remainsa challenge, since reaction products diffuse away from the enzymeand make it difficult to recover catalytically active phage–enzymes.We have recently described a selection methodology in whichthe reaction substrate (and eventually the reaction product)is anchored on calmodulin-tagged phage–enzymes by meansof a calmodulin binding peptide. Phage displaying a catalyticactivity are physically isolated by means of affinity reagentsspecific for the product of reaction. In this study, we investigatedthe efficiency of selection for catalysis by phage display,using a ligase (the Escherichia coli biotin ligase BirA) andan endopeptidase (the rat trypsin His57     

Inhibition of lipid oxidation by encapsulation of emulsion droplets within hydrogel microspheres

The purpose of this study was to assess whether the oxidation of polyunsaturated lipids could be inhibited by encapsulating them within protein-rich hydrogel microspheres (size range 1-100 μm). Filled hydrogel microspheres were fabricated as follows: (i) high methoxy pectin, sodium caseinate, and casein-coated lipid droplets were mixed at pH 7, (ii) the mixture was acidified (pH 5), (iii) casein was cross-linked using transglutaminase, (iv) the pH was adjusted to pH 7. Samples were stored in the dark at 55 °C and were monitored for lipid hydroperoxide formation and headspace propanal. Oxidation of fish oil (1% vol/vol) in the microspheres was compared with that in oil-in-water emulsions stabilised by either sodium caseinate or Tween 20. Emulsions stabilised by Tween 20 oxidised faster than either microspheres or emulsions stabilised by casein, while microspheres and the casein stabilised emulsion showed similar oxidation rates. Results highlight the natural antioxidant properties of food proteins.