‘Lipase and protease production of dairy Penicillium sp. on milk‐protein‐based solid substrates’

The lipolytic and proteolytic activity of Penicillium camemberti PC TT033 and Penicillium roqueforti PR G3, cultured on the whey solids or simulated cheese media, were compared under several pH reaction conditions. Lipolytic activity was higher when both strains had been cultured on the whey medium than on the simulated cheese medium, whereas proteolytic activity was less influenced by the culture medium. The relationship between the reaction pH and these enzyme activities was dependent on the culture medium, which suggested that the expression level and balance of isozyme rely on the culture substrate.     

Design of AVS/RS under group constraint

Distribution centers, which are essential to our society, are becoming increasingly important. Although the efficiency of an AVS/RS (Autonomous vehicle storage and retrieval system) makes it a promising system, there is still much to be studied regarding its effective use and design. This paper proposes a method of designing AVS/RSs through a quantitative consideration of group constraint by utilizing probability theory. The proposed algorithm also deals with the problem of designing the sizes of several buffers, and considers the effect of load fluctuations, which are important for real-world operation, by use of queuing network theory. Effectiveness of the proposed algorithm is shown with simulations of various number of groups, those of various safety factors for fluctuations, and those of various evasion rates for buffer overflow. The design results are shown to be much different from those which do not consider those factors. The results prove that the proposed method provides an accurate quantitative assessment of the AVS/RS’s constraints.     

Microcoaxial electrode for in vivo nitric oxide measurement

Nitric oxide (NO) is a gaseous mediator involved in various physiological phenomena, such as vasorelaxation and neurotransmission. Investigation of local cellular responses of NO production in vivo and in vitro requires a measurement method with a high spatial resolution. For selective NO measurement, we therefore developed a microcoaxial electrode whose tip diameter is less than 10 microm. Calibration using various concentrations of NO (0.1-1.0 microM) showed that the electrode has good linearity (r = 0.99) and its detection limit is 0.075 microM (S/N = 3). We verified the applicability of this electrode to in vivo and in vitro local measurement NO released from bovine aortic cultured endothelial cells (BAECs) stimulated by acetylcholine (ACh). After the addition of ACh, a transient increase in NO concentration was detected by the electrode. In the presence of NG-nitro-L-arginine methyl ester (L-NAME), a putative NO synthase inhibitor, NO release (peak NO concentration) from RAECs was significantly less than that in the absence of L-NAME (0.18 +/- 0.04 microM vs 0.47 +/- 0.13; P < 0.01). After removal of L-NAME, NO release partially recovered (0.39 +/- 0.10 microM). In conclusion, the microcoaxial electrode was successfully applied to direct and continuous NO measurement in biological systems.     

A Single Mutation at the Ferredoxin Binding Site of P450 Vdh Enables Efficient Biocatalytic Production of 25‐Hydroxyvitamin D3

Vitamin D₃ hydroxylase (Vdh) from Pseudonocardia autotrophica is a cytochrome P450 monooxygenase that catalyzes the two‐step hydroxylation of vitamin D₃ (VD₃) to produce 25‐hydroxyvitamin D₃ (25(OH)VD₃) and 1α,25‐dihydroxyvitamin D₃ (1α,25(OH)₂VD₃). These hydroxylated forms of VD₃ are useful as pharmaceuticals for the treatment of conditions associated with VD₃ deficiency and VD₃ metabolic disorder. Herein, we describe the creation of a highly active T107A mutant of Vdh by engineering the putative ferredoxin‐binding site. Crystallographic and kinetic analyses indicate that the T107A mutation results in conformational change from an open to a closed state, thereby increasing the binding affinity with ferredoxin. We also report the efficient biocatalytic synthesis of 25(OH)VD₃, a promising intermediate for the synthesis of various hydroxylated VD₃ derivatives, by using nisin‐treated Rhodococcus erythropolis cells containing Vdh_T107A. The gene‐expression cassette encoding Bacillus megaterium glucose dehydrogenase‐IV was inserted into the R. erythropolis chromosome and expressed to avoid exhaustion of NADH in a cytoplasm during bioconversion. As a result, approximately 573 μg mL^?1 25(OH)VD₃ was successfully produced by a 2 h bioconversion.     

Albumin stabilizes (–)‐epigallocatechin gallate in human serum: Binding capacity and antioxidant property

(–)‐Epigallocatechin gallate (EGCg) is the major component of green tea and is known to show strong biological activity, although it can be easily oxidized under physiological conditions. In this study, we indicate that EGCg is stable in human serum and that human serum albumin (HSA) stabilizes EGCg under aerobic condition. Although EGCg is usually decomposed within 1 h in aqueous solution at neutral pH, EGCg in serum and phosphate buffer (pH 7.4) containing HSA was stable over 1 h, even at neutral and slightly alkaline pH. Under these conditions, EGCg binds to HSA non‐covalently. The sulfhydryl group acts as an antioxidant for EGCg oxidation. Incubation of EGCg with HSA is accompanied by the oxidation of a free sulfhydryl group in HSA. These results suggest that the antioxidant property and the binding capacity of HSA contribute to the stabilization of EGCg in human serum.     

Comprehensive analysis of minimum grain size in pure aluminum using friction stir processing

A friction stir processing (FSP) technique has been developed for use with aluminum and magnesium alloys, with the goal of high-strain-rate processing. In this study, we treat the microstructures of aluminum samples, of three levels of purity, manipulating their grain sizes, making them finer, using FSP. Grain sizes decreased with an increase in the Zener-Hollomon parameter, at all purity levels. However, the recrystallized grain of FSP-ed ultra-high-purity (99.999%) aluminum was particularly large, compared to the grain sizes of lower-purity (99% and 99.99%) aluminum, when subjected to the same processing conditions. The grain size reached a certain minimum value at high-strain-rates of processing, which we report for each purity level herein.     

On an implicit stress-calculation algorithm for a multidimensional constitutive law using a skeleton curve

In this paper, we present an implicit stress-calculation algorithm for a multidimensional constitutive law using a one-dimensional skeleton curve and a hysteresis curve. To consider the hysteretic behavior of soils, one-dimension skeleton curves (e.g., the Hardin–Drnevich model, Ramberg–Osgood (RO) model, and general hyperbolic equation (GHE) model) are widely used. In a multidimensional analysis, there are several methods for extending a constitutive law using a skeleton curve into multiple dimensions. However, because these methods are suitable for an explicit integration stress-calculation scheme, a calculated stress does not satisfy the function of skeleton curve when a large incremental strain is imposed in one step. In this paper, to resolve this problem, a nonlinear stress–strain relation using a skeleton curve and an implicit stress-calculation algorithm is presented. In addition, we show an example of an application of the GHE/RO model to the proposed algorithm.