Studies on the Dehydrogenative Polymerizations of Monolignol β-glycosides. Part 3: Horseradish Peroxidase–Catalyzed Polymerizations of Triandrin and Isosyringin

Abstract

Horseradish peroxidase–catalyzed dehydrogenative polymerizations of the p-hydroxyphenyl monolignol glucoside (triandrin (1P)) and the syringyl monolignol glucoside (isosyringin (1S)) resulted in the formation of water-soluble lignin-like polymers (DHPs). The polymerization of 1P gave highly polymerized DHPs in high yields as did previously reported polymerization of the guaiacyl monolignol glucoside (isoconiferin (1G)). It was shown that the hydrophilic D-glucose units of 1G and 1P contribute to a marked increase in the molecular weights of the resulting DHPs. On the other hand, the homogeneous phase polymerization of 1S, similar to the polymerization of sinapyl alcohol, gave DHPs with extremely low molecular masses in poor yields. Structural characterization indicated that the DHPs from 1P and 1S were lignin-like polymers containing glucosidic units on their sidechains. It was also confirmed that D-glucosyl units introduced onto the γ-position of monolignols do not significantly affect the electrochemical oxidizability and the kinetics of the HRP-catalyzed initial monomer consumption.     

How temporal cues can aid colour constancy

Colour constancy assessed by asymmetric simultaneous colour matching usually reveals limited levels of performance in the unadapted eye. Yet observers can readily discriminate illuminant changes on a scene from changes in the spectral reflectances of the surfaces making up the scene. This ability is probably based on judgements of relational colour constancy, in turn based on the physical stability of spatial ratios of cone excitations under illuminant changes. Evidence is presented suggesting that the ability to detect violations in relational colour constancy depends on temporal transient cues. Because colour constancy and relational colour constancy are closely connected, it should be possible to improve estimates of colour constancy by introducing similar transient cues into the matching task. To test this hypothesis, an experiment was performed in which observers made surface-colour matches between patterns presented in the same position in an alternating sequence with period 2 s or, as a control, presented simultaneously, side-by-side. The degree of constancy was significantly higher for sequential presentation, reaching 87% for matches averaged over 20 observers. Temporal cues may offer a useful source of information for making colour-constancy judgements.     

Sex Pheromone Components of the Pear Fruit Moth, <Emphasis Type="Italic">Acrobasis pyrivorella</Emphasis> (Matsumura)

We analyzed the sex pheromone of the pear fruit moth, Acrobasis pyrivorella, by means of gas chromatography–electroantennographic detection (GC-EAD) and GC–mass spectrometry. Two EAD-active compounds were detected in the pheromone gland extract of females. They were identified as (Z)-9-pentadecenyl acetate (Z9-15:OAc) and pentadecyl acetate (15:OAc). The amounts per female gland (mean ± standard error) of these compounds were 12.9 ± 2.8 and 0.8 ± 0.1 ng, respectively. Synthetic Z9-15:OAc (300 μg) attracted conspecific males in field trapping experiments. When 15:OAc (21 μg; 7% of Z9-15:OAc quantity) was added, the number of males trapped increased significantly. Catch in traps baited with the mixture of these compounds was greater than that in traps baited with 1–3-day-old virgin females. We, therefore, conclude that Z9-15:OAc and 15:OAc are sex pheromone components of this species.     

Xylanase from Marine Filamentous Fungus Pestalotiopsis sp. AN-7 Was Activated with Diluted Salt Solution Like Brackish Water

The genus Pestalotiopsis are endophytic fungi that have recently been identified as cellulolytic system producers. We herein cloned a gene coding for a xylanase belonging to glycoside hydrolase (GH) family 10 (PesXyn10A) from Pestalotiopsis sp. AN-7, which was isolated from the soil of a mangrove forest. This protein was heterologously expressed by Pichia pastoris as a host, and its enzymatic properties were characterized. PesXyn10A was produced as a glycosylated protein and coincident to theoretical molecular weight (35.3 kDa) after deglycosylation by peptide-NfF-glycosidase F. Purified recombinant PesXyn10A exhibited maximal activity at pH 6.0 and 50 °C, and activity was maintained at 90 % at pH 5.0 and temperatures lower than 30 °C for 24 h. The substrate specificity of PesXyn10A was limited and it hydrolyzed glucuronoxylan and arabinoxylan, but not β-glucan. The final hydrolysis products from birchwood xylan were xylose, xylobiose, and 1,2³-α-D-(4-O-methyl-glucuronyl)-1,4-β-D-xylotriose. The addition of metallic salts (NaCl, KCl, MgCl₂, and CaCl₂) activated PesXyn10A for xylan degradation, and maximal activation by these divalent cations was approximately 160 % at a concentration of 5 mM. The thermostability of PesXyn10A significantly increased in the presence of 50 mM NaCl or 5 mM MgCl₂. The present results suggest that the presence of metallic salts at a low concentration, similar to brackish water, exerts positive effects on the enzyme activity and thermal stability of PesXyn10A.     

The inhibitory effects of components from a brown alga,Eisenia arborea,on degranulation of mast cells and eicosanoid synthesis

The anti-allergic effect of an edible brown alga, Eisenia arborea, was investigated by determining the anti-degranulation activity on RBL cells or KU812 cells and the inhibitory effects on enzymatic activities involved in eicosanoid (leukotoriene and prostaglandin) synthesis. When the stimulated cells were treated with a M/C (methanol:chloroform = 1:2, v/v) extract of the alga, the histamine release of the cells was suppressed in a dose-dependent manner, and eicosanoid release from the cells was also strongly inhibited. Furthermore, the M/C extract and six phlorotannins (eckol, 6,6′-bieckol, 6,8′-bieckol, 8,8′-bieckol, phlorofucofuroeckol-A, phlorofucofuroeckol-B) derived from the extract inhibited enzymatic activities responsible for eicosanoid synthesis. This is the first report of the enzymatic activity inhibition by the phlorotannins 6,6′-bieckol, 6,8′-bieckol and PFF-B. Since histamine and eicosanoids are known to mediate the allergic reaction, inhibition of these mediators and enzymatic activities could be responsible for the anti-allergic effect of the alga.     

Optimal aerobic cultivation method for 1,4-dihydroxy-2-naphthoic acid production by Propionibacterium freudenreichii ET-3

To investigate the effects of oxygen supply on Propionibacterium freudenreichii ET-3 metabolism and 1,4-dihydroxy-2-naphthoic acid (DHNA) production in detail, the strain was cultured by switching from anaerobic condition to aerobic condition at 72 h (termed anaerobic-aerobic switching culture hereafter) employing different oxygen transfer rates (OTRs) in the range of 0.08-0.90 mg/(l.h). It was found that a 0.08 mg/(l.h) OTR could not change the metabolism or improve the DHNA production of P. freudenreichii ET-3. When the OTR was in the range of 0.23-0.66 mg/(l.h), propionate, which inhibits DHNA production significantly, was consumed during the aerobic phase. Final DHNA concentration increased to 0.22 mM, irrespective of OTR. When the OTR was 0.90 mg/(l.h), a sudden increase in dissolved oxygen (DO) concentration during the aerobic phase resulted in a sudden decrease in DHNA concentration. To attenuate the stresses caused by propionate and oxygen exposure, we designed an optimal cultivation in which the anaerobic and aerobic phases were repeated three times alternately. As a result, propionate concentration was maintained below the level that inhibits DHNA production, and no DO concentration was detected throughout the culture. The final DHNA concentration in this culture was 0.33 mM, which is 2.7-fold that in the anaerobic culture and 1.5-fold that in the anaerobic-aerobic switching culture.     

Rapid enumeration of low numbers of moulds in tea based drinks using an automated system

Aseptically prepared cold drinks based on tea have become popular worldwide. Contamination of these drinks with harmful microbes is a potential health problem because such drinks are kept free from preservatives to maximize aroma and flavour. Heat-tolerant conidia and ascospores of fungi can survive pasteurization, and need to be detected as quickly as possible. We were able to rapidly and accurately detect low numbers of conidia and ascospores in tea-based drinks using fluorescent staining followed by an automated counting system. Conidia or ascospores were inoculated into green tea and oolong tea, and samples were immediately filtered through nitrocellulose membranes (pore size: 0.8 μm) to concentrate fungal propagules. These were transferred onto potato dextrose agar and incubated for 23 h at 28 °C. Fungi germinating on the membranes were fluorescently stained for 30 min. The stained mycelia were counted selectively within 90 s using an automated counting system (MGS-10LD; Chuo Electric Works, Osaka, Japan). Very low numbers (1 CFU/100 ml) of conidia or ascospores could be rapidly counted, in contrast to traditional labour intensive techniques. All tested mould strains were detected within 24 h while conventional plate counting required 72 h for colony enumeration. Counts of slow-growing fungi (Cladosporium cladosporioides) obtained by automated counting and by conventional plate counting were close (r² = 0.986). Our combination of methods enables counting of both fast- and slow-growing fungi, and should be useful for microbiological quality control of tea-based and also other drinks.     

Performance of single‐ and double‐effect operable mechanical vapor recompression desalination system adaptable to variable wind energy

This paper deals with the development and operation of a mechanical vapor recompression (MVR) desalination system with improved energy efficiency in harnessing wind energy, which is non‐dispatchable. Its design, construction, and operation details are presented in this paper. Especially, the main focus of developing the system was on the operation of the system in conjunction with variable loads of new and renewable power sources, in particular, varying wind power. That is, the present work has been carried out to assess the feasibility of its operation in light of capacity modulation to match the power generated under various wind speeds. Optimal operation modes of the system were studied, in which single‐ and double‐effect operations were analyzed for their improvement in energy efficiency. The compression ratio of the proposed MVR system was 1.55 at an inverter speed of 55 Hz, which agreed well with its design value. Operation of the main heat exchanger remained stable within the limits of its operable range, although the temperature differences in the main heat exchanger did not remain constant because of the pressure variations in the evaporator. The daily freshwater yield was between 28 and 51 tons. The power consumption per ton of freshwater produced was about 43 kW for a single effect and about 23 kW for a double effect, which is about twice as efficient.