Expression of a gene encoding chitinase (pCA 8 ORF) from Aeromonas sp. no. 10S-24 in Escherichia coli and enzyme characterization

A gene encoding chitinase from Aeromonas sp. no. 10S-24 was expressed using pTrc99A in Escherichia coli JM 105 which yielded a 5-fold higher activity than when pUC19 was used. Three different truncated enzymes (SA-1, SA-2 and SA-3) were obtained after purification. Their isoelectric points were 7.0, 6.9, and 6.7, respectively. The enzymes showed two optimum pHs, 4.0 and 7.0, when incubated with ethylene glycol chitin as the substrate, and were stable over a wide pH range (3.0-9.0). The optimum temperature was 60 degrees C and the enzymes were stable up to 50 degrees C. The chitinases exhibited wide substrate specificities for chitin-related compounds.     

Direct ethanol production from barley beta-glucan by sake yeast displaying Aspergillus oryzae beta-glucosidase and endoglucanase

Three beta-glucosidase- and two endoglucanase-encoding genes were cloned from Aspergillus oryzae, and their gene products were displayed on the cell surface of the sake yeast, Saccharomyces cerevisiae GRI-117-UK. GRI-117-UK/pUDB7 displaying beta-glucosidase AO090009000356 showed the highest activity against various substrates and efficiently produced ethanol from cellobiose. On the other hand, GRI-117-UK/pUDCB displaying endoglucanase AO090010000314 efficiently degraded barley beta-glucan to glucose and smaller cellooligosaccharides. GRI-117-UK/pUDB7CB codisplaying both beta-glucosidase AO090009000356 and endoglucanase AO090010000314 was constructed. When direct ethanol fermentation from 20 g/l barley beta-glucan as a model substrate was performed with the codisplaying strain, the ethanol concentration reached 7.94 g/l after 24 h of fermentation. The conversion ratio of ethanol from beta-glucan was 69.6% of the theoretical ethanol concentration produced from 20 g/l barley beta-glucan. These results showed that sake yeast displaying A. oryzae cellulolytic enzymes can be used to produce ethanol from cellulosic materials. Our constructs have higher ethanol production potential than the laboratory constructs previously reported.     

Estimation of daily intake of phenols in hospital meal samples

Daily intakes of 12 phenols which are possible endocrine disruptors were estimated in hospital meals from 2000 to 2001. 4-Nonylphenol (4-NP (mix)) and bisphenol A (BPA) were detected at levels of 5.0 to 19.4 ng/g and 0.2 to 1.1 ng/g, respectively. 4-tert-Butylphenol, 4-n-pentylphenol, 4 -tert-pentylphenol, 4-n-hexylphenol, 4-n-heptylphenol and 4-tert-octylphenol were detected at levels of 0.1 to 2.4 microg/g. The daily intakes of 4-NP (mix) and BPA were 5.8 microg/day and 0.42 microg/day, respectively. The daily intakes of other phenols were less than 1 microg/day.     

A Method for Reducing the Dead‐Time Voltage and Impedance Voltage in a Series Voltage Compensator

Many research groups are developing series voltage compensators. In a series converter, since a transformer is used in series in the power system, the power system current flows into the voltage source inverter through the transformer. The inverter current, which is determined by the transformation ratio, gives rise to an error voltage that consists of a dead‐time voltage and an impedance voltage. The error voltage is generated even when the reference voltage is zero. This paper describes the mechanism by which the error voltage occurs and proposes a method for reducing the error voltage. © 2013 Wiley Periodicals, Inc. Electr Eng Jpn, 186(3): 85–93, 2014; Published online in Wiley Online Library ( wileyonlinelibrary.com ). DOI 10.1002/eej.22333     

Raw Starch Digestion by Bacillus polymyxa beta-Amylase

Unlike plant ß-amylases, Bacillus polymyxa ß-amylase digests raw starch at a significant rate. The action of this bacterial ß-amylase on raw starch is stimulated greatly by the presence of the debranching enzyme pullulanase.     

Relationship between local structure and stability in hen egg white lysozyme mutant with alanine substituted for glycine

We prepared five mutant lysozymes in which glycines whose dihedralangles are located in the region of the left-handed helix, Gly49,Gly67, Gly71, Gly102 and Gly117, were mutated to an alanineresidue. From analyses of their thermal stabilities using differentialscanning calorimetry, most of them were more destabilized thanthe native lysozyme, except for the G102A mutant, which hasa stability similar to that of the native lysozyme at pH 2.7.As for the destabilized mutant lysozymes, their X-ray crystallographicanalyses showed that their global structures did not changebut that the local structures changed slightly. By examiningthe dihedral angles at the mutation sites based on X-ray crystallographicresults, it was found that the dihedral angles at these mutationsites tended to adopt favorable values in a Ramachandran plotand that the extent and direction of their shifts from the originalvalue had similar tendencies. Therefore, the change in dihedralangles may be the cause of the slight local structural changesaround the mutation site. On the other hand, regarding the mutationof G102A, the global structure was almost identical with thatof the native structure but the local structure was drasticallychanged. Therefore, it was suggested that the drastic localconformational change might be effective in releasing the unfavorableinteraction of the native state at the mutation site.     

Steric Effect on the Catalytic Performance of the Selective Oxidation of Alcohols Over Novel Crystalline Mo–V–O Oxide

The Mo–V–O crystalline oxide with novel pore structure and consisting uniform six- and seven-member rings on the a–b plane is investigated for the selective oxidation of alcohols of different steric hindrance in liquid phase. The research target is to correlate catalytic activity with the pore structure of the crystal. Specially, substituted pyridines are employed as probe molecules to study poison effect which is closely related to the steric hindrance. As a result, the oxidation of benzyl alcohol, 1-hexanol and cyclohexanol produces aldehydes or ketones as main products. The oxidation of substrates with methyl groups on the carbon next to alcohol group mainly affords dehydrated products as olefins. The catalytic results with adding substituted pyridines in the oxidation of benzyl alcohol, 1-hexanol and 2-hexanol suggest that the active sites are located around the pore area, and are reachable by pyridine, not by substituted pyridines, such as 2-methylpyridine, 2-ethylpyridine and 2, 6-dimethylpyridine. Competitive adsorption on active sites between pyridine and benzyl alcohol remarkably decreases catalytic activity, which 2, 6-dimethylpyridine affects slightly. We have discussed that the adsorption-activation model of substrate is greatly dependent on its steric hindrance.     

Periplasmic expression of human growth hormone via plasmid vectors containing the {lambda}PL promoter: use of HPLC for product quantification

The influence of different factors acting on Escherichia coliperiplasmic expression of recombinant human growth hormone (hGH)in shake flask cultures has been investigated. Bacterial vectorscontaining the phage     

Effects of pressure and temperature on dyeing acrylic fibres with basic dyes in supercritical carbon dioxide

The dyeing of acrylic fibre with CI Basic Blue 47 has been investigated using an ammonium carboxylate perfluoropolyether as an auxiliary in order to perform the reverse micellar system in supercritical carbon dioxide media. The basic dye was satisfactorily solubilised in the interior of the reverse micelle in the system, and dyeability in the supercritical carbon dioxide media was excellent, even in the absence of a retardant and/or an acid. The optimum proton-rich condition for dyeing of acrylic fabric is performed in reverse micelle aggregates and the proton-rich water pool is suitable for dye uptake by the fibre even without the presence of additives. Changes in glass transition temperature of acrylic fibre in the carbon dioxide media also influence the dyeing behaviour of acrylic fabric.