Cells of <Emphasis Type="BoldItalic">Candida utilis</Emphasis> for <Emphasis Type="BoldItalic">in vitro</Emphasis> (<Emphasis Type="BoldItalic">R</Emphasis>)-phenylacetylcarbinol production in an aqueous/octanol two-phase reactor

(R)-Phenylacetylcarbinol (PAC), a pharmaceutical precursor, was produced from benzaldehyde and pyruvate by pyruvate decarboxylase (PDC) of Candida utilis in an aqueous/organic two-phase emulsion reactor. When the partially purified enzyme in this previously established in vitro process was replaced with C. utilis cells and the temperature was increased from 4 to 21 °C, a screen of several 1-alcohols (C4–C9) confirmed the suitability of 1-octanol as the organic phase. Benzyl alcohol, the major by-product in the commercial in vivo conversion of benzaldehyde and sugar to PAC by Saccharomyces cerevisiae, was not formed. With a phase volume ratio of 1:1 and 5.6 g C. utilis l⁻¹ (PDC activity 2.5 U ml⁻¹), PAC levels of 103 g l⁻¹ in the octanol phase and 12.8 g l⁻¹ in the aqueous phase were produced in 15 h at 21 °C. In comparison to our previously published process with partially purified PDC in an aqueous/octanol emulsion at 4 °C, PAC was produced at a 4-times increased specific rate (1.54 versus 0.39 mg U⁻¹ h⁻¹) with simplified catalyst production and reduced cooling cost. Compared to traditional in vivo whole cell PAC production, the yield on benzaldehyde was 26% higher, the product concentration increased 3.9-fold (or 6.9-fold based on the organic phase), the productivity improved 3.1-fold (3.9 g l⁻¹ h⁻¹) and the catalyst was 6.9-fold more efficient (PAC/dry cell mass 10.3 g g⁻¹).*Dedicated with gratitude to Prof. Dr. Franz Lingens – “Theo”.     

Evaluation of the bioremediatory potential of several species of the red alga <Emphasis Type="BoldItalic">Porphyra</Emphasis> using short-term measurements of nitrogen uptake as a rapid bioassay

Rates of inorganic nitrogen uptake by three Northeast US and three Asian species of Porphyra were compared in short-term incubations to evaluate potential for longer term and larger scale examination of bioremediation of nutrient-loaded effluents from finfish aquaculture facilities. The effects of nitrogen (N) species and concentration, temperature, acclimation history, and irradiance were investigated. Uptake rates increased ca. nine-fold from 20 to 150 μM N. Nitrate and ammonium uptake occurred at similar rates. Irradiance had a strong effect, with uptake at 40 μmol photons m⁻² s⁻¹only 55% of uptake at 150 μmol photons m⁻² s⁻¹. N-replete tissue took up inorganic nitrogen at rates that averaged only 60% of nutrient-deprived tissue. Although there were species (P. amplissima > (P. purpurea = P. umbilicalis)) and temperature effects (10 ^°C>5 ^°C>15 ^°C), interactions among factors indicated that individual species be considered separately. Overall, P. amplissima was the best Northeast US candidate. It took up ammonium at faster rates than other local species at 10 and 15 ^°C, two temperatures that fall within the expected range of industrial conditions for finfish operations.     

Cooperation of xanthophyll cycle with water-water cycle in the protection of photosystems 1 and 2 against inactivation during chilling stress under low irradiance

The xanthophyll cycle and the water-water cycle had different functional significance in chilling-sensitive sweet pepper upon exposure to chilling temperature (4 °C) under low irradiance (100 µmol m⁻² s⁻¹) for 6 h. During chilling stress, effects of non-photochemical quenching (NPQ) on photosystem 2 (PS2) in dithiothreitol (DTT) fed leaves remained distinguishable from that of the water-water cycle in diethyldithiocarbamate (DDTC) fed leaves. In DTT-fed leaves, NPQ decreased greatly accompanied by visible inhibition of the de-epoxidized ratio of the xanthophyll cycle, and maximum photochemical efficiency of PS2 (F_v/F_m) decreased markedly. Thus the xanthophyll cycle-dependent NPQ could protect PS2 through energy dissipation under chilling stress. However, NPQ had a slighter effect on photosystem 1 (PS1) in DTT-fed leaves than in DDTC-fed leaves, whereas effects of the water-water cycle on PS1 remained distinguishable from that of NPQ. Inhibiting superoxide dismutase (SOD) activity increased the accumulation of , the oxidation level of P700 (P700⁺) decreased markedly relative to the control and DTT-fed leaves. Both F_v/F_m and NPQ changed little in DDTC-fed leaves accompanied by little change of (A+Z)/(V+A+Z). This is the active oxygen species inducing PS1 photoinhibition in sweet pepper. The water-water cycle can be interrupted easily at chilling temperature. We propose that during chilling stress under low irradiance, the xanthophyll cycle-dependent NPQ has the main function to protect PS2, whereas the water-water cycle is not only the pathway to dissipate energy but also the dominant factor causing PS1 chilling-sensitivity in sweet pepper.This research was supported by the State Key Basic Research and Development Plan of China (G1998010100), the Natural Science Foundation of China (30370854), and the open project from Key Lab of Crop Biology of Shandong Province.     

Purification and characterization of naringinase from a newly isolated strain of <Emphasis Type="Italic">Aspergillus niger</Emphasis> 1344 for the transformation of flavonoids

Summary An extracellular naringinase (an enzyme complex consisting of α-L-rhamnosidase and β-D-glucosidase activity, EC 3.2.1.40) that hydrolyses naringin (a trihydroxy flavonoid) for the production of rhamnose and glucose was purified from the culture filtrate of Aspergillus niger 1344. The enzyme was purified 38-fold by ammonium sulphate precipitation, ion exchange and gel filtration chromatography with an overall recovery of 19% with a specific activity of 867 units per mg of protein. The molecular mass of the purified enzyme was estimated to be about 168 kDa by gel filtration chromatography on a Sephadex G-200 column and the molecular mass of the subunits was estimated to be 85 kDa by sodium dodecyl sulphate-Polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme had an optimum pH of 4.0 and temperature of 50 °C, respectively. The naringinase was stable at 37 °C for 72 h, whereas at 40 °C the enzyme showed 50% inactivation after 96 h of incubation. Hg²⁺, SDS, p-chloromercuribenzoate, Cu²⁺ and Mn²⁺ completely inhibited the enzyme activity at a concentration of 2.5–10 mM, whereas, Ca²⁺, Co²⁺ and Mg²⁺ showed very little inactivation even at high concentrations (10–100 mM). The enzyme activity was strongly inhibited by rhamnose, the end product of naringin hydrolysis. The enzyme activity was accelerated by Mg²⁺ and remained stable for one year after storage at −20 °C. The purified enzyme preparation successfully hydrolysed naringin and rutin, but not hesperidin.     

Phytoplankton primary production in a eutrophic cooling water pond

The seasonal variation of phytoplankton photosynthesis was measured with ¹⁴C-method in a warmed ice-free pond in central Finland. Simultaneously with in situ measurements the photosynthesis was also measured in an incubator with different water temperatures and constant light (ca. 16 W m^–2). The total annual photosynthesis was 57.2 C m^–2 a^–1. The portion of the winter and spring production of the annual photosynthesis was 18.4%, that of the autumn production ws 17.4%. Thus 64.3% of the total annual phytoplankton photosynthesis occurred in the three summer months. The range of the daily integrated photosynthesis per unit area was 1.9—563 mg C m^–2d^–1. The photosynthetic rate per unit chlorophyll a varied in situ from 0.94 to 33.1 mg C (mg chl. a)^–1 d^–1. The highest value was measured in the beginning of July and the lowest in mid-January. The photosynthetic rate increased in situ exponentially with increasing water temperature. In the incubator the highest photosynthetic rate values were also found in July and August (at+20 °C) when the phytoplankton population was increasing and the minimum values occurred after every diatom maximum both in spring and autumn. Light was a limiting factor for photosynthesis from September to Mid-January, low water temperature was a limiting factor from late January through May. The efficiency of the photosynthesis varied between 0.1 and 0.7% of P.A.R. According to the incubator experiments the Q₁₀ values for the photosynthesis were 2.45 and 2.44 for the winter population between 1 and 10° C and for the summer population between 5 and 15° C, respectively, but the Q₁₀ values decrease at the higher temperatures. The main effect of the warm effluents on the yearly photosynthesis was the increase of production in spring months due to the lack of ice cover. However, the increase of total annual phytoplankton photosynthesis was only ca. 10–15%, because the water temperature was during the spring months below 10° C.     

Physico-chemical characterization of exomannan from <Emphasis Type="Italic">Rhodotorula acheniorum</Emphasis> MC

The batch fermentation of Rhodotorula acheniorum MC on a culture medium containing 5% sucrose, mineral salts and yeast extract at 26 °C for 96 h, with aeration at 0.75 v/v/m and agitation at 500 rev min ⁻¹ resulted in the synthesis of an exopolysaccharide (6.2 g l ⁻¹) which formed two fractions upon precipitation. The fractions were purified to a carbohydrate content of 98.2% for fraction I and 87.3% for fraction II. Mannose was the main monosaccharide component in a 92.8% concentration in fraction I and a 90.6% concentration in Fraction II. The exopolysaccharide was thus a mannan. The gel chromatograms confirmed the chemical composition of both fractions. The molecular weight of mannan I was 310 kD, whereas that of mannan II was 249 kD. The mannan I intrinsic viscosity [η]=6.23 dl g⁻¹ was higher than that of mannan II [η]=2.73 dl g⁻¹. The water-binding capacity of the mannan samples was established within the 1.2–3.5 g g⁻¹ range. The multiplicative model [η]=387.22. D_r^−0.1913. T^−1.095. C^1.814 describing the effect of the velocity gradient D_r, the exomannan concentration C and the temperature T on the dynamic viscosity values η of polymer solutions was obtained.     

Physiological acclimation to light in Chara intermedia nodes

In this study we investigated the ability of Chara intermedia to acclimate to different irradiances (i.e. “low-light” (LL): 20–30 μmol photons m⁻² s⁻¹ and “high-light” (HL): 180–200 μmol photons m⁻² s⁻¹) and light qualities (white, yellow and green), using morphological, photosynthesis, chlorophyll fluorescence and pigment analysis.Relative growth rates increased with increasing irradiance from 0.016 ± 0.003 (LL) to 0.024 ± 0.005 (HL) g g⁻¹ d⁻¹ fresh weight and were independent of light quality. A growth-based branch orientation towards high-light functioning as a mechanism to protect the plant from excessive light was confirmed. It was shown that the receptor responsible for the morphological reaction is sensitive to blue-light.C. intermedia showed higher oxygen evolution (up to 10.5 (HL) vs. 4.5 (LL) nmol O₂ mg Chl⁻¹ s⁻¹), photochemical and energy-dependent Chl fluorescence quenching and a lower Fv/Fm after acclimation to HL. With respect to qP, the acclimation of the photosynthetic apparatus depended on light quality and needed the blue part of the spectrum for full development. In addition, pigment composition was influenced by light and the Chl a/Car and Antheraxanthin (A) + Zeaxanthin (Z)/Violaxanthin (V) + A + Z (DES) ratios revealed the expected acclimation behaviour in favour of carotenoid protection under HL (i.e. decrease of Chl a/Car from 3.41 ± 0.48 to 2.30 ± 0.35 and increase of DES from 0.39 ± 0.05 to 0.87 ± 0.03), while the Chl a/Chl b ratios were not significantly affected. Furthermore it was shown that morphological light acclimation mechanisms influence the extent of the physiological modifications.     

Regulation of nitrogenase activity by light in the azolla-anabaena symbiosis

Nitrogenase activity and the rate of photosynthesis were measured simultaneously in Azolla by a continuous gas flow system. The mode of interaction between light, photosynthesis and nitrogenase activity was analysed.Nitrogenase activity dropped off when either Azolla plants or the cyanobiont Anabaena were transferred from light to dark. This decline was immediate and was independent of length or intensity of the prior light phase. Reillumination restored nitrogenase activity.Nitrogenase activity did not depend on the rate of photosynthesis at light intensities below 10 μE m⁻² s⁻¹. Its activity was saturated at 200 μE m⁻² s⁻¹ while CO₂ fixation was saturated at a light intensity of 850 μE m⁻² s⁻¹. Azolla photosynthetic activity followed the absorption spectrum of chlorophyll a, while nitrogenase activity markedly increased between 690 and 710 nm. Inhibition of photosynthesis by DCMU was accompanied by an increase in nitrogenase activity. These results suggest direct light regulation of nitrogenase activity in Azolla independent of CO₂ fixation, and a possible inhibition of nitrogenase activity by the oxygen produced in photosynthesis.     

Inhibition of photosynthesis by chilling in moderate light: a comparison of plants sensitive and insensitive to chilling

Photosynthetic activity, in leaf slices and isolated thylakoids, was examined at 25° C after preincubation of the slices at either 25° C or 4° C at a moderate photon flux density (PFD) of 450 mol·m^–2·s^–1, or at 4° C in the dark. The plants used wereSpinacia oleracea L.,Cucumis sativus L. andNerium oleander L. which was acclimated to growth at 20° C or 45° C. The plants were grown at a PFD of 550 mol·m^–2·s^–1. Photosynthesis, measured as CO₂-dependent O₂ evolution, was not inhibited in leaf slices from any plant after preincubation at 25° C at a moderate PFD or at 4° C in the dark. However, exposure to 4° C at a moderate PFD induced an inhibition of CO₂-dependent O₂ evolution within 1 h inC. sativus, a chilling-sensitive plant, and in 45° C-grownN. oleander. The inhibition in these plants after 5 h reached 80% and 40%, respectively, and was independent of the CO₂ concentration but was reduced at O₂ concentrations of less than 3%. Methyl-viologen-dependent O₂ exchange in leaf slices from these plants was not inhibited. There was no photoxidation of chlorophyll, in isolated thylakoids, or any inhibition of electron transport at photosystem (PS)II, PSI or through both photosystems which would account for the inhibition of photosynthesis. The conditions which inhibit photosynthesis in chilling-sensitive plants do not cause inhibition inS. oleracea, a chilling-insensitive plant, or in 20° C-grownN. oleander. The CO₂-dependent photosynthesis, measured at 5° C, was reduced to about 3% of that recorded at 25° C in chilling-sensitive plants but only to about 30% in the chilling-insensitive plants. Methyl-viologen-dependent O₂ exchange, measured at 5° C, was greater than 25% of the activity at 25° C in all the plants. The results indicate that the mechanism of the chilling-induced inhibition of photosynthesis does not involve damage to PSII. That inhibition of photosynthesis is observed only in the chilling-sensitive plants indicates it is related, in some way, to the disproportionate decrease in photosynthetic activity in these plants at chilling temperatures.Abbreviations Chl chlorophyll - DPIPH reduced form of 2,6-dichlorophenol-indophenol - DMQ 2,5-dimethyl-p-benzoquinone - MV methyl viologen - 20°-oleander Nerium oleander grown at 20° C - 45°-oleander N. oleander grown at 45° C - PFD photon flux density (photon fluence rate) - PSI and PSII photosystem I and II, respectively     

Exogenous expression of the wheat chloroplastic fructose-1,6-bisphosphatase gene enhances photosynthesis in the transgenic cyanobacterium, <Emphasis Type="BoldItalic">Anabaena</Emphasis> PCC7120

The paper reports a study on the genetic regulation of photosynthesis by introducing the gene encoding wheat chloroplastic fructose-1,6-bisphosphatase (FBPase) into the cyanobacterium Anabaena PCC7120. The gene was RT-PCR amplified from wheat and modified by replacement of the 5′-terminal encoding sequence with optimal and A/T-rich codons to promote prokaryotic expression. The resultant FBPase gene was ligated downstream of the strong promoter, PpsbA of expression vector pRL-439, then inserted into of shuttle vector pDC-08. The resulting shuttle expression vector (pDC-fbp) was transferred into the filamentous, heterocystour cyanobacterium, Anabaena PCC7120, by the tri-parental conjugation transfer method. Protein expression of FBPase in the transgenic Anabaena was 126.5% higher than in wild type cells, and the enzyme activity of transgenic cells was 1.41-fold higher than that of wild type cells. Under atmospheric conditions of 360 μmol mol⁻¹ CO₂, Anabaena cells overexpressing the FBPase gene further showed increases in net photosynthesis (117.2%) and true photosynthesis (122.5%) as compared to wild type cells. In addition, transgenic Anabaena grew faster and contained more Chl a than did wild type cells. Together, these results indicate that introduction of the wheat chloroplastic FBPase gene into Anabaena increase photosynthesis and cell growth; furthermore, these trends were more evident under stress condition (higher CO₂ concentration). This is the first report of enhanced photosynthesis in cyanobacteria expressing genes from higher plants.     

Abundance and growth response of microalgae at Megalon Embolon solar saltworks in northern Greece: An aquaculture prospect

There is continuous interest in many countries in maintaining and manipulating the rich ecological value of hypersaline ecosystems for aquaculture. The Megalon Embolon solar saltworks (northern Greece) were studied in sites of increasing salinity of 60–144 ppt to evaluate Dunaliella salina abundance and microalgal composition, in relation to physical and chemical parameters. Cluster and ordination analyses were performed based on the biotic and abiotic data matrices. Using fresh aliquots from 60 and 140 ppt salinity waters, phytoplankton performance was appraised with flask cultures in the laboratory by varying the inorganic PO₄-P concentration at 23 °C and 30 °C. At the saltworks, among the most abundant microalgae identified were species of the genera Dunaliella, Chlamydomonas, Amphora, Navicula, and Nitzschia. Dunaliella salina populations were predominant comprising 5–22% of the total microalgal assemblages during spring, but only 0.3–1.0% during the summer, when grazing by Artemia parthenogenetica and Fabrea salina was intense. D. salina cell density in April–July was in the range of 0.4–12.5 × 10⁶ L⁻¹ with typical densities of 1.5–4.5 × 10⁶ L⁻¹. Overall, microalgal densities were high in salinities of ≥100 ppt when inorganic-P concentrations were ≥0.20 mg L⁻¹ within saltworks waters. Multivariate analysis of species abundance showed that algal growth responses were primarily related to variation in salinity and inorganic-P concentrations, but also to NO₃-N concentration. In the laboratory, experiments indicated effective fertilization and denser microalgal growth under high inorganic PO₄-P applications (4.0 and 8.0 mg L⁻¹) at 60 ppt salinity and 23 °C. The lower PO₄-P applications (0.6–2.0 mg L⁻¹) were more effective at 60 ppt salinity and 30 °C. At 140 ppt salinity, microalgal growth response was less obvious at any of the corresponding phosphorus concentrations or temperatures. In both salinity experiments, Dunaliella salina bloomed easily and was predominant among the microalgae. Our observations indicate that Dunaliella salina populations and the overall rich microalgal profile of the saltworks, along with their performance in laboratory mono–and mixed cultures hold promise for mass cultivation within the M. Embolon saltworks basins.     

Interaction Among Copper Toxicity,Temperature and Salinity on the Population Dynamics of <Emphasis Type="Italic">Brachionus Rotundiformis</Emphasis> (Rotifera)

Heavy metals may interact with ecological factors such as temperature, food level and salinity, causing both mortality and reduced reproduction in organisms. Among different heavy metals, copper compounds are commonly used for eliminating algal blooms in aquaculture tanks. At certain concentrations, copper is toxic to rotifers. In the present work, we evaluated the combined effects of salt concentrations (2.5 and 5.0 g l⁻¹ NaCl), copper levels (0, 0.03125, 0.0625, 0.125 and 0.25 mg l⁻¹ as CuCl₂) and two temperatures (20 and 25 °C) on the population growth of B. rotundiformis using Chlorella as the algal food (at 0.5 × 10⁶ cells ml⁻¹ for every 24 h). Regardless of salinity and temperature, copper at concentrations as low as 0.03 mg l⁻¹ had an adverse effect on the population growth of rotifers and above 0.125 mg l⁻¹, the populations did not grow. The effect of the toxicant on B. rotundiformis was more severe at 25° than at 20 °C at lower salinity. In general, we observed peak densities of rotifers around day 12 at 20 °C but 6–8 days earlier at 25 °C. Peak population densities of B. rotundiformis in the controls at the salinity of 2.5 g l⁻¹ ranged from 90 to 180 ind. ml⁻¹, depending on temperature; at a salinity of 5.0 g l⁻¹, these were lower. The population growth rates, r, in our study varied from +0.31 to –0.12 depending on the test conditions. There was a significant impact of temperature, salinity and toxicity level on the population growth rate of B. rotundiformis. Our results suggested that even narrow changes in salinity could negatively influence the toxicity of heavy metal on the population growth rates of B. rotundiformis.     

Aspects of the Cryobiology of the Intertidal Harpacticoid CopepodTigriopus brevicornis(O. F. Müller)

Cold-resistance studies of marine invertebrates have concentrated on intertidal sedentary organisms, which are often subjected to subzero air temperatures in winter. Mobile rock pool inhabitants have been rarely studied because such habitats are thought to buffer environmental variation. However, it is not uncommon for small upper-shore rock pools (2 by 1 cm) to become completely frozen. Such supralittoral habitats are subject to extreme physicochemical fluctuations especially in salinity (0 to 300‰) and temperature (−1 to +32°C) due to evaporation and dilution. The dominant invertebrate in such habitats is the harpacticoid copepodTigriopus brevicornis.Aspects of the cryobiology ofT. brevicorniswere investigated using differential scanning calorimetry (DSC). Thermograms obtained from DSC allowed determinations of freeze-onset (supercooling point, SCP), melt-onset, and melt-peak (melting point, MP) temperatures, together with estimation of the proportion of water freezing in the samples. The effects of acclimation salinity, temperature, starvation, and reproductive state on these cryobiological parameters were investigated. Acclimation to increasing salinity depressed the SCP, with the highest salinity (70‰) producing the lowest SCP, melt-onset, and MP temperatures at −27.5, −15.2, and −9.5°C respectively. The highest acclimation temperature (20°C) produced the lowest SCP (−23.4°C). Starvation significantly increased the SCP, melt-onset, and MP temperatures in comparison to fed individuals acclimated to the same salinity. The presence of eggs or ovaries in individual copepods elevated the SCP compared to nongravid females and males. LT₅₀studies showed that acclimation to high salinity improved the ability ofT. brevicornisto survive in frozen seawater. Seventy parts per thousand acclimated individuals had an LT₅₀of 64.9 h compared with just 1.4 h for 5‰ acclimated individuals in frozen seawater at −5°C. The study shows that the cold-resistance capabilities ofT. brevicorniscan be affected by several different factors, and the link between the osmoconforming nature of this species and its cold resistance is discussed.     

In Vitro Selection of High Temperature Zn2+-Dependent DNAzymes

In vitro selection of Zn²⁺-dependent RNA-cleaving DNAzymes with activity at 90°C has yielded a diverse spool of selected sequences. The RNA cleavage efficiency was found in all cases to be specific for Zn²⁺ over Pb²⁺, Ca²⁺, Cd²⁺, Co²⁺, Hg²⁺, and Mg²⁺. The Zn²⁺-dependent activity assay of the most active sequence showed that the DNAzyme possesses an apparent Zn²⁺-binding dissociation constant of 234 μM and that its activity increases with increasing temperatures from 50–90°C. A fit of the Arrhenius plot data gave E_a = 15.3 kcal mol⁻¹. Surprisingly, the selected Zn²⁺-dependent DNAzymes showed only a modest (∼3-fold) activity enhancement over the background rate of cleavage of random sequences containing a single embedded ribonucleotide within an otherwise DNA oligonucleotide. The result is attributable to the ability of DNA to sustain cleavage activity at high temperature with minimal secondary structure when Zn²⁺ is present. Since this effect is highly specific for Zn²⁺, this metal ion may play a special role in molecular evolution of nucleic acids at high temperature.     

Purification and characterization of lignin peroxidases from <Emphasis Type="Italic">Penicillium decumbens</Emphasis> P6

Summary Peroxidases are essential enzymes in biodegradation of lignin and lignite which have been investigated intensively in the white-rot fungi. This is the first report of purification and characterization of lignin peroxidase from Penicillium sp. P6 as lignite degradation fungus. The results indicated that the lignin peroxidase of Penicillium decumbens P6 had physical and chemical properties and a N-terminal amino acid sequence different from the lignin peroxidases of white-rot fungi. The lignin peroxidase was isolated from a liquid culture of P. decumbens P6. This enzyme had a molecular weight of 46.3 KDa in SDS-PAGE and exhibited greater activity, temperature stability and wider pH range than those previously reported. The isolation procedure involved (NH₄)₂SO₄ precipitation, ion-exchange chromatography on DEAE-cellulose and CM-cellulose, gel filtration on Sephadex G-100, and non-denaturing, discontinuous polyacrylamide gel electrophoresis. The K _m and V _max values of this enzyme using veratryl alcohol as substrate were 0.565 mmol L ⁻¹ and 0.088 mmol (mg protein) ⁻¹ min ⁻¹ respectively. The optimum pH of P6 lignin peroxidase was 4.0, and 70.6 of the relative activity was remained at pH 9.0. The optimum temperature of the enzyme was 45 °C.     

Chromate reduction by <Emphasis Type="Italic">Bacillus megaterium</Emphasis> TKW3 isolated from marine sediments

Summary Bacillus megaterium strain TKW3 was isolated from multiple-metal-contaminated marine sediments of Tokwawan, Hong Kong SAR. This facultative aerobe utilized arabinose, mannitol, N-acetylglucosamine, maltose, caprate, citrate, butyrate or lactate as the sole source of carbon and energy for growth.B. megaterium TKW3 reduced highly toxic and soluble Cr⁶⁺ (as CrO₄²⁻) into almost non-toxic and insoluble Cr³⁺ under aerobic conditions. Complete reduction of 0.20 mM Cr⁶⁺ by B. megaterium TKW3 was achieved within 360 h. Initial Cr⁶⁺ concentration below 0.90 mM or inoculum less than 10⁷ cells ml⁻¹ did not have significant effect on ⁶⁺ reduction, while the residue Cr⁶⁺ concentration was the lowest at 10⁷ cells ml⁻¹. Cr⁶⁺ reduction by this strain was inhibited by high levels of NaCl (55%). B. megaterium TKW3 was also resistant to other oxyanions including 0.34 mM Cr₂O₇²⁻ 0.32 mM AsO₄³⁻, 0.58 mM SeO₃²⁻ and 0.53 mM SeO₄²⁻, and reduced soluble Se⁴⁺ (as SeO₃²⁻) to insoluble red amorphous Se⁰. B. megaterium TKW3 might have potential application in bioremediation of Cr-laden sediments associated with other oxyanions.     

Regulation of gamete release in the economic brown seaweed <Emphasis Type="Italic">Hizikia fusiforme</Emphasis> (Phaeophyta)

Gamete release is an essential event in artificial seeding of the economic brown seaweed, Hizikia fusiforme. Mass egg release occurred in the dark, with few eggs being discharged in the light. Release of eggs was elicited with eight practical salinity units (one PSU ≡ 1 g sea salts l⁻¹) and was inhibited by salinity levels >32 PSU. Egg release was optimal at 23 °C, and was decreased by 72% in agitated seawater compared to unstirred seawater. Inhibitors of photosynthesis and ions channels suppressed egg release, indicating that this process was physiologically associated with photosynthetic activity and ion transport.     

Factors Affecting Swimming Speed in the Rotifer <Emphasis Type="Italic">Brachionus plicatilis</Emphasis>

This study examines the swimming speed in amictic females of Brachionus plicatilis in laboratory cultures. Five different stages were examined: recently hatched females, juveniles, adult non-ovigerous females, ovigerous females with 1 attached egg and ovigerous females with 2 attached eggs. We tested the speed at two temperatures, 15 °C and 25 °C, and two feeding conditions, presence and absence of microalgal cells. An automated motion analysis system was used to measure speed which was then video recorded. Swimming speed (μm s⁻¹) increased with increasing body size. There was a slight decrease in the speed of adult females as the number of attached eggs increased. Swimming activity was higher at 25 °C than at 15 °C and in the absence of food than if microalgae were present. Average values under the different experimental conditions ranged between 500 μm s⁻¹ for the recently hatched and fed females and 1500 μm s⁻¹ for the adult non-ovigerous females in the absence of microalgae. Mass-specific swimming speed decreased with body mass increase.     

Life history and growth in culture of the endemic New Zealand kelp <Emphasis Type="Italic">Lessonia variegata</Emphasis> J. Agardh in response to differing regimes of temperature,photoperiod and light

Lessonia variegata J. Agardh (Laminariales, Phaeophyta) is endemic to New Zealand, where it occurs in subtidal kelp forests on wave exposed coasts in the North, South and Stewart Islands. This is the first account of the growth in culture and life history of L. variegata. Microscopic gametophytes alternate with macroscopic sporophytes, characteristic of members of the order Laminariales. The life history was completed in culture within 14 days under growth conditions of 12 °C, 12:12 (L:D) and 15 °C, 15:9 (L:D). Maximum growth of sporophytes occurred at 15 °C, 15:9, and slowest growth at 10 °C, 9:15. Under low light conditions (8–9 μmol photon m⁻² s⁻¹) filamentous growth of gametophytes predominated, and both the proportion of gametogenesis and the growth of sporophytes after 30 days was much reduced from equivalent cultures grown under conditions of higher light (16–17.5 μmol photon m⁻² s⁻¹). Interest in this species relates both to its potential for commercial utilisation as well as in the role it plays in coastal rocky reef ecosystems.     

Purification and characterisation of NAD+-dependent xylitol dehydrogenase from Fusarium oxysporum

An NAD⁺-dependent xylitol dehydrogenase (XDH) from Fusarium oxysporum, a key enzyme in the conversion of xylose to ethanol, was purified to homogeneity and characterised. It was homodimeric with a subunit of M _r 48 000, and pI 3.6. It was optimally active at 45 °C and pH 9–10. It was fully stable at pH 6–7 for 24 h and 30 °C. K _m values for d-xylitol and NAD⁺ were 94 mM and 0.14 mM, respectively. Mn²⁺ at 10 mM increased XDH activity 2-fold and Cu²⁺ at 10 mM inhibited activity completely.