The influence of the ion content of whey on the pH-activity profile of the β-galactosidase from Aspergillus oryzae

The pH-dependence of the reaction kinetics of lactase (β-galactosidase) from Aspergillus oryzae in different reaction media is presented in terms of a two-parameter model. The lactase from A. oryzae seems to have replaced the A. niger lactase on the market owing to a better activity/price ratio and may be utilised for lactose hydrolysis in acid as well as in neutral milk products. Its pH optimum is around pH 4.5. However, in the neutral pH-range its activity depended strongly on the salt content of the substrate solution. For example, its activity in whey (pH 6.5) fell to only 30% of its expected activity in a pure lactose solution at the same pH. The whey effect was the same for both soluble and immobilised lactase. The two parameter kinetic model, which included a term for competitive product inhibition gave excellent agreement with experimental data, and may thus be useful for the prediction of reactor performance with this enzyme.     

Quantitative evaluation of generator power control effect of FACTS controllers for power system stabilization

The advancement of power electronics technologies has significantly developed the power system stabilizing controllers. Quantitative as well as qualitative evaluation of their effectiveness in power systems is a matter of great importance for the feasibility investigation of these apparatus. In this paper, the possible control region of FACTS controllers with series and/or shunt configuration in a single machine to infinite bus system is formulated in the powerangle curve with a set of algebraic equations. The effectiveness of TCPST (Thyristor‐Controlled Phase Shifting Transformer), SSSC (Static Synchronous Series Compensator), and TCSC (Thyristor‐Controlled Series Compensator) for the improvement of the transient stability is evaluated quantitatively as a numerical example. The correctness of the proposed method has been confirmed by analysis based on the electromagnetic transients simulation with a detailed system model. © 2001 Scripta Technica, Electr Eng Jpn, 138(3): 43–51, 2002     

Sensorless control of synchronous reluctance motors using on‐line parameter identification method not affected by position estimation accuracy

This paper presents a novel on‐line parameter identification method for sensorless control of Synchronous Reluctance Motors (SynRMs). Although conventional sensorless control methods based on mathematical models usually need some complex measurements of motor parameters in advance, the proposed identification method does not require them and can be realized on‐line. The proposed method identifies motor parameters under sensorless control, so rotor position and velocity cannot be used to identify these parameters. However, the proposed method does not need rotor position and velocity, and identified parameters are not affected by these estimation errors. The sensorless control using identified motor parameters is realized, and the effectiveness of the proposed method is verified by experimental results. © 2006 Wiley Periodicals, Inc. Electr Eng Jpn, 155(3): 62–69, 2006; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/eej.20258     

A novel robust current control approach using adaptive line enhancer for three‐phase current‐source active power filter

An effective system control method is presented for applying a three‐phase current‐source PWM converter with a deadbeat controller to active power filters (APFs). In the shunt‐type configuration, the APF is controlled such that the current drawn by the APF from the utility is equal to the current harmonics and reactive current required for the load. To attain the time‐optimal response of the APF supply current, a two‐dimensional deadbeat control scheme is applied to APF current control. Furthermore, in order to cancel both the delay in the two‐dimensional deadbeat control scheme and the delay in DSP control strategy, an Adaptive Line Enhancer (ALE) is introduced in order to predict the desired value three sampling periods ahead. ALE has another function of bringing robustness to the deadbeat control system. Due to the ALE, settling time is made short in a transient state. On the other hand, total harmonic distortion (THD) of source currents can be minimized compared to the case where ideal identification of the controlled system can be made. The experimental results obtained from the DSP‐based APF are also reported. The compensating ability of this APF is very high in accuracy and responsiveness although the modulation frequency is rather low. © 2004 Wiley Periodicals, Inc. Electr Eng Jpn, 150(1): 50–61, 2005; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/eej.20014     

Yeast exo-β-glucanases can be used as efficient and readily detectable reporter genes in Saccharomyces cerevisiae

Yeast exo-1,3-β-glucanases are secretable proteins whose function is basically trophic and may also be involved in cell wall glucan hydrolytic processes. Since fluorescein di(β-D -glucopyranoside) is a fluorogenic substrate detectable and quantifiable by flow cytometry, it was used for testing the ability of the EXG1 gene product of Saccharomyces cerevisiae and its homologous gene in Candida albicans to function as reporter genes. These open reading frames were coupled to different promoters in multicopy plasmids, and exoglucanase activity quantified at flow cytometry. Exoglucanases were found to be useful tools for the study of promoter regions in S. cerevisiae. This technique has the advantage over other reporter gene systems—such as β-galactosidase fusions—that it does not require permeabilization of yeast cells and therefore it allows the recovery of viable cells—by sorting—after flow cytometry analysis.     

Yeast sequencing reports. Isolation and sequence analysis of a gene from the linear DNA plasmid pPAC1–2 of Pichia acaciae that shows similarity to a killer toxin gene of Kluyveromyces lactis

The toxin-encoding linear plasmid systems found in Pichia acaciae and Kluyveromyces lactis yeasts appear to be quite similar, both in function and structural organization. By Southern hybridization, a linear plasmid of P. acaciae, pPac1–2, was found to hybridize to the second open reading frame (ORF2) of K. lactis plasmid pGKL1, known to encode the α and β subunits of the K. lactis toxin. A 1·7 kbp segment of pPac1–2 DNA was cloned, sequenced and shown to contain four regions of strong homology to four similarly oriented regions of K. lactis ORF2. This 1·7 kbp fragment also contained an ORF of 1473 bp that could encode a protein of ～ 55·8 kDa. Like the α subunit gene of K. lactis ORF2, a very hydrophobic region occurs at the N-terminus, perhaps representing a signal sequence for transport out of the cell. Unlike K. lactis ORF2, however, the encoded polypeptide is much smaller and lacks a recognizable domain common to chitinases. The structure of a toxin that includes the translation product of this P. acaciae ORF would likely be quite different from that of the K. lactis toxin. Analysis of the upstream region of the P. acaciae ORF revealed an upstream conserved sequence identical to that found before ORFs 8 and 9 of pGKL2. A possible hairpin loop structure, as has been described for each of the four K. lactis pGKL1 ORFs, was found just upstream of the presumed start codon. The similarity of the promoter-like elements found in the linear plasmid genes of these diverse yeasts reinforces the idea of the existence of a unique, but highly conserved, expression system for these novel plasmids. The sequence has been deposited in the GenBank data library under Accession Number U02596.     

Electrical conductivity in iodine-doped ethyl cellulose

The electrical conductivity of solution-grown ethyl cellulose (EC) films, 5–30 μm thick, has been studied in the sandwich configuration (metal–EC–metal) as a function of iodine concentration from 0.5 to 5.0 wt% ratio. The studies were conducted in the temperature range 333–383 K, while the field was varied over the range (3.0–5.5) × 10⁴V/cm. Aluminium was used as the lower electrode, while the upper electrode was of Al, Ag, Cu, Au or Sn. Certain transient effects such as a large burst of current immediately after the application of field were observed. An attempt was made to identify the nature of the current by comparing the observed dependence on electric field, electrode material and temperature with the respective characteristic features of the existing theories of electrical conduction. The results show that the electrical conduction follows Ohm's law at lower fields, while at higher fields, space-charge limited current (SCLC) was observed. It was also found that Richardson–Schottky emission was responsible, to some extent, for the transport of charge carriers in the polymer. The conductivity of the films increased on doping with iodine. The dopant molecules are considered to act as additional trapping centes and provide links between the polymer molecules in the amorphous region, thus resulting in the formation of charge transfer complexes.     

Purification and renaturation of recombinant human lymphotoxin (tumour necrosis factor beta) expressed in Escherichia coli as inclusion bodies

High level expression of recombinant human tumour necrosis factor β (rh TNF-β) in Escherichia coli results in the formation of two portions of protein, namely soluble active protein and insoluble protein which is inactive and aggregates in the form of inclusion bodies (IBs). In this study, a procedure for purification and renaturation of rh TNF-β from inclusion bodies has been designed and verified experimentally with a product purity of more than 90% and a recovery of about 30%. The procedure includes washing of IBs with specific wash buffer (Triton X-100/EDTA/lysozyme/PMSF), their solubilization with 8 mol dm^?3 alkaline urea, purification with ion-exchange columns, refolding with renaturation buffer and finally concentration and desalination with an ultrafiltration membrane. The characteristics of the renatured protein were identical with those of purified protein from the soluble fraction as demonstrated by (1) SDS-PAGE, (2) cytotoxic activity on mouse L929 cells, (3) N-terminal amino acid sequence, and (4) gel filtration chromatography.