Enhanced glucan formation of filamentous fungi by effective mixing,oxygen limitation and fed-batch processing

Summary Glucan formation ofSchizophyllum commune andSclerotium glucanicum were investigated. Process data obtained during batch cultivation are presented. Glucan release can be improved by oxygen limitation. Thus, growth and glucan release are influenced by oxygen in opposite ways. Possible pathways of this oxygen-dependent regulation are discussed. A draft-tube/propeller system, rushtonturbine-, fan- and helicon-ribbon-impeller as well as a fundaspi and intermig agitator were tested. The 4-bladed fan impeller withd ^*=0.64 yielded the best results, since effective bulk mixing is much more important than bubble break up (micromixing) with regard to this system. Fed-batch cultivation always resulted in higher rates of glucan formation than the batch process.     

Some factors influencing the proportion of periplasmic hepatitisB virus pre-S2 antigen in the recombinant yeastHansenula polymorpha

Summary A central composite design (CCD) was used to evaluate, for the purpose of future process optimization, the influence of pH, yeast extract and ammonium chloride concentrations on the proportion of periplasmic hepatitisB pre-S2 antigen in the recombinant yeastHansenula polymorpha. Each factor was tested at five levels, and a second order polynomial model for the proportion of periplasmic antigen was fitted to the treatment combinations. pH showed the greatest effect: the proportion of periplasmic antigen was greatly increased at the higher pH levels. At the higher pH levels used, the proportion of periplasmic antigen was enhanced by a high concentration of ammonium chloride. Additional experiments have confirmed both the validity of the selected model and the optimal conditions found. A significant correlation was found between the proportion of periplasmic antigen and the total yield of antigen. These results indicated that is should be possible to modulate the distribution of the pre-S2 antigen between the periplasm and the cytoplasm of the yeast.     

The Exact Distribution of the Anderson-Kannemann Statistic

The Anderson-Kannemann test is a rank test for treatment effects in a randomized block design with K treatments and N blocks. In this paper, an algorithm for computing the exact distribution of the Anderson-Kannemann test statistic under the null hypothesis is deduced. Then, the exact distribution is compared with the asymptotic χ²-distribution, and it is shown that the exact distribution is approximated fairly well by the asymptotic distribution. Tables of the exact distribution are given for K = 3, N = 3(1)15; K = 4, N = 3(1)11; K = 5, N = 3(1)7; and K = 6, N = 3(1)5.     

Optimization of integrated alkaline–extrusion pretreatment of barley straw for sugar production by enzymatic hydrolysis

In this work, an integrated one-step alkaline–extrusion process was tested as pretreatment for sugar production from barley straw (BS) biomass. The influence of extrusion temperature (T) and the ratio NaOH/BS dry matter (w/w) (R) into the extruder on pretreatment effectiveness was investigated in a twin-screw extruder at bench scale. A 2³ factorial design of experiments was used to analyze the effect of process conditions [T: 50–100 °C; R: 2.5–7.5% (w/w)] on composition and enzymatic digestibility of pretreated substrate (extrudate). The optimum conditions for a maximum glucan to glucose conversion were determined to be R = 6% and T = 68 °C. At these conditions, glucan yield reached close to 90% of theoretical, while xylan conversion was 71% of theoretical. These values are 5 and 9 times higher than that of the untreated material, which supports the great potential of this one-step combined pre-treatment technology for sugar production from lignocellulosic substrates. The absence of sugar degradation products is a relevant advantage over other traditional methods for a biomass to ethanol production process since inhibitory effect of such product on sugar fermentation would be prevented.     

Preparation and evaluation of soluble epoxide hydrolase inhibitors with improved physical properties and potencies for treating diabetic neuropathic pain

Soluble epoxide hydrolase (sEH), a novel therapeutic target for neuropathic pain, is a largely cytosolic enzyme that degrades epoxy-fatty acids (EpFAs), an important class of lipid signaling molecules. Many inhibitors of sEH have been reported, and to date, the 1,3-disubstituted urea has the highest affinity reported for the sEH among the central pharmacophores evaluated. An earlier somewhat water soluble sEH inhibitor taken to the clinic for blood pressure control had mediocre potency (both affinity and kinetics) and a short in vivo half-life. We undertook a study to overcome these difficulties, but the sEH inhibitors carrying a 1,3-disubstituted urea often suffer poor physical properties that hinder their formulation. In this report, we described new strategies to improve the physical properties of sEH inhibitors with a 1,3-disubstituted urea while maintaining their potency and drug-target residence time (a complementary in vitro parameter) against sEH. To our surprise, we identified two structural modifications that substantially improve the potency and physical properties of sEH inhibitors carrying a 1,3-disubstituted urea pharmacophore. Such improvements will greatly facilitate the movement of sEH inhibitors to the clinic.     

Computational Modeling of Protein Stability: Quantitative Analysis Reveals Solutions to Pervasive Problems

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