Carbamoylethylation of guar gum

Carbamoylethylation of guar gum was carried out with acrylamide in presence of sodium hydroxide under different reaction conditions. Variables studied were concentration of sodium hydroxide, acrylamide, guar gum as well as reaction temperature and time. The nitrogen content, carboxyl content and total ether content were determined. Rheological properties of carbamoylethyl guar gum solutions showed non-Newtonian pseudoplastic behavior regardless of the %N. At a constant rate of shear, the apparent viscosity of carbamoylethyl guar gum solutions decreases with the increase in %N of the product.     

夏蜡梅属花粉形态的研究

本文首次报道了夏蜡梅(Calycanthus chinensis)花粉的扫描电镜观察结果,并与西美国蜡梅(C.occidentalis)、美国蜡梅(C.floridus)的作了比较,表明前者较为原始,后者比较进化,夏蜡梅属(Calycanthus L.)可能以夏蜡梅起源于东亚(中国),与从细胞学、形态学等得到的结果一致。鉴于夏蜡梅与西美国蜡梅、美国蜡梅在许多性状上的较大差异,作者认为以Sinocalycanthus chinensis (Cheng et S.Y.Chang) Cheng et S.Y.Chang为模式种的单型属Sinocalycanthus Cheng et S.Y.Chang也许应予确认。     

Morphology and germination characteristics of the cysts of Chattonella ovata (Raphidophyceae), a novel red tide flagellate in the Seto Inland Sea, Japan

To elucidate the mechanism of bloom outbreaks of Chattonella ovata (Raphidophyceae), we investigated the cysts of C. ovata and succeeded in finding them from the bottom sediments of Hiroshima Bay. The morphology of the cysts was mostly hemispherical in shape, with a diameter of ca. 30 μm and height of ca. 20 μm. The cysts were usually adhering to solid materials, such as diatom frustules, yellow-greenish in color and had several dark brown grains. The cyst wall was smooth and had no ornamentation. Because the morphological characteristic of the cysts was in general agreement with those of Chattonella antiqua and Chattonella marina, it was difficult to differentiate the cysts of these three species. Germination of the cysts of C. ovata was observed at temperatures from 17.5 to 30 °C, but not at 15 °C or below. The number of the germinated cysts increased with increasing temperature and the optimum temperature for germination was 30 °C. Although cysts of C. antiqua and C. marina germinated at temperatures from 15 to 30 °C, optimum temperature of germination was 22.5 °C. The lower limit and optimum temperatures for germination of C. ovata cysts was higher than for C. antiqua and C. marina. The role of cysts in the population dynamics of C. ovata is discussed.     

Lattice deformation and thermal stability of crystals in spider silk

The X-ray diffraction of dragline silks, produced by Nephila and Cyrtophora spiders, were measured by synchrotron radiation in their original states or in situ during stretching and heating. Nephila pilipes spiders construct a two-dimensional orb web that must be rebuilt in one or 2 days, but Cyrtophora spiders form a three-dimensional tent web that can exist for several weeks in a tropical forest. Diffraction patterns of N. pilipes and Cyrtophora draglines resemble each other. Crystals of two kinds are identified in these draglines; one is aligned parallel to the silk direction and another is less oriented. The less oriented crystal in Cyrtophora dragline is aligned better than that in N. pilipes dragline, which generates about three times stronger diffract intensity. Crystals in N. pilipes and C. moluccensis dragline silks have remarkable thermal stability. Equatorial reflections remain undiminished until 350 and 450 °C for N. pilipes and C. moluccensis, respectively. In contrast, the meridional reflections S and (0 0 2), which are parallel to the silk thread, disappear at a temperature less than 100 °C for C. moluccensis but remain for Nephila up to 100 °C. Meridional reflections S and (0 0 2) shift to a smaller angle during stretching, whereas equatorial reflections remain constant in a range 1.0–1.3 times the original length. The position of the S reflection shifts rapidly in the first 10% of elongation from the original length but remains constant during subsequent stretching, whereas the (0 0 2) reflection shifts rapidly during the first 5% elongation from the original length and continues to shift subsequently. In contrast, the features of N. pilipes dragline alter insignificantly during stretching. Examination of the composition of amino acids of the draglines of N. pilipes and C. moluccensis indicates that a dragline of N. pilipes contains more glycine, but much less alanine, than that of C. moluccensis.     

Critical thermal maximum and body water loss in first instar larvae of three Cetoniidae species (Coleoptera)

Critical thermal maximum (CT_max) and body water losses were measured in first instar larvae of Gnorimus nobilis, Osmoderma eremita (Trichiinae) and Cetonischema aeruginosa (Cetoniinae) when air temperature was increased gradually (0.5 °C/min) from 20 °C to the critical thermal maximum (CT_max), in dry air (near 0% R.H.).

The CT_max was significantly lower in O. eremita (45.6±0.7 °C) than in G. nobilis (48.5±0.6) and C. aeruginosa (51.4±0.9 °C).

An increase of 10 °C (30–40 °C) induced a 2-fold increase of the water loss in C. aeruginosa and O. eremita (Q₁₀=2.10±0.12 and 2.13±0.20, respectively). In the range from 40 to 45 °C to CT_max a strong increase of the water loss was observed in O. eremita and C. aeruginosa, respectively. Body water losses were significantly lower in C. aeruginosa than in O. eremita and G. nobilis over the range 20 °C—CT_max; no significant difference occurred between G. nobilis and O. eremita.     

钠盐胁迫对河西走廊荒漠绿洲过渡带典型一年生草本植物种子萌发的影响

研究不同浓度盐分(NaCl和NaHCO₃,0、40、80、120、160和200 mmol·L^-1)对6种河西走廊荒漠绿洲边缘人工林典型一年生草本植物(禾本科:狗尾草、虎尾草和小画眉草;藜科:雾冰藜、刺沙蓬和蒙古虫实)种子在当年秋季和次年春季的萌发和复萌的影响。结果表明: NaCl和NaHCO₃对6种植物种子的萌发有显著影响,NaHCO₃对种子萌发的抑制作用大于NaCl。NaHCO₃浓度达到160 mmol·L^-1时,6种植物基本不萌发或萌发率较低;NaCl浓度达到200 mmol·L^-1时,6种植物仍能够保持相对较高的萌发率。种子当年秋季萌发率(2.8%～20.0%)远低于次年春季萌发率(21.7%～81.6%),秋季复萌率(3.3%～20.0%)低于春季复萌率(5.0%～41.1%),当年生种子秋季多保持休眠,次年春季休眠减弱。6种一年生草本植物种子的耐盐性为:虎尾草＞狗尾草＞刺沙蓬＞雾冰藜＞蒙古虫实＞小画眉草。     

更新方式对亚热带森林土壤溶液可溶性有机质数量及化学结构的影响

采用负压法对福建省三明市亚热带常绿阔叶林中米槠次生林(BF)、米槠人促更新林(RF)、米槠人工林(CP) 0～15、15～30、30～60 cm土层土壤溶液可溶性有机质(DOM)的浓度及光谱学特征进行研究.结果表明: 土壤溶液可溶性有机碳(DOC)浓度整体趋势为RF>CP>BF,而可溶性有机氮(DON)则为米槠人工林最高;且 DOC 和 DON 在表层(0～15 cm)土壤浓度皆显著高于底层(30～60 cm).芳香化指数大小为RF>CP>BF,且整体为表层较高.米槠人工林表层土壤以荧光强度高的短波峰(320 nm)为特征峰,表明其易分解物质含量高,腐殖化程度较低;而米槠人促更新林表层土壤则以宽平的中长波峰(380 nm)为特征峰,说明其腐殖化程度较高,有助于土壤肥力的储存.此外,30～60 cm深层DOM特性几乎不受森林更新方式的影响.     

Direct heat injury of roots of woody swamp species

1. 1.Three month-old seedlings of Taxodium distichum, Nyssa aquatica, Cephalanthus occidentalis and cuttings of Salix nigra were acclimated to simulated natural swamp conditions (ambient temperature, saturated soil) and then used to assess direct high temperature injury of root tissue.

2. 2.Electrolyte leakage from excised root tissue exposed for 30 min to temperatures ranging from 30 to 66°C was used to assess cellular injury.

3. 3.The relationship between leakage and temperature was sigmoidal for each species.

4. 4.Inflection point temperatures on the response curves, ranged from 45.4 to 51.0°C, were species-specific, and indicated differences in thermal tolerance of root membranes.

5. 5.Root of C. occidentalis and N. aquatica were more heat tolerant than roots of T. distichum and S. nigra.

Conversion of Japanese red cedar (Cryptomeria japonica) into a feed for ruminants by white-rot basidiomycetes

The potential of five white-rot fungi to convert Japanese red cedar (Cryptomeria japonica) into feed for ruminants was determined. Pleurotus ostreatus (ATCC 66376), Pholiota nameko (IFO 30373), Dichomitus squalens (CBS 432.34), Lentinula edodes (IFO 6654) and Ceriporiopsis subvermispora (ATCC 90467) were inoculated into chips of cedar sapwood. L. edodes was cultured at 25 °C, and the other fungi at 28 °C, for 4, 8, 12, or 20 weeks. The in vitro organic matter (OM) digestibility (IVOMD) in cedar wood cultured without fungus were between 0.047 and 0.068, while it was elevated to 0.446 by culturing with C. subvermispora and to 0.281 by culturing with L. edodes for 20 weeks. In contrast, the IVOMD were 0.200, or lower, in cedar wood cultured with P. ostreatus, P. nameko, or D. squalens. The in vitro gas production (IVGP) in cedar wood cultured with P. ostreatus, P. nameko, or D. squalens was 37 ml/g OM, or lower, while that in cedar not inoculated with fungus was between 4 and 17 ml/g OM. In contrast, the IVGP in cedar wood cultured with C. subvermispora for 20 weeks increased to 107 ml/g OM, and that in cedar wood cultured with L. edodes increased to 58 ml/g OM. Lignin degradability in cedar wood cultured with C. subvermispora and L. edodes for 20 weeks were 0.578 and 0.288, respectively. These changes in IVOMD and IVGP demonstrate that a selective white-rot fungus, C. subvermispora has the ability to convert cedar wood into feed for ruminants, although further increase is required before the cultured cedar wood would have widespread feed potential.     

Immobilization of wood-rotting fungi laccases on modified cellulose and acrylic carriers

Extracellular laccases produced by three different wood-rotting fungi, Cerrena unicolor, Heterobasidion annosum and Trametes versicolor, were immobilized via covalent bonds formation on DEAE-Granocel 500, CM-Granocel 500, and acrylic carriers. Out of the tested carriers, only the DEAE-Granocel 500, which was activated by divinyl sulphone appeared to be a suitable matrix for the expression of enzymic activity. Only one laccase of all the tested enzymes produced by C. unicolor showed the best binding to the carrier and a satisfactory enzymic activity. The immobilized laccase exhibited the highest enzymic activity at pH 5.2 and it was more resistant to thermal denaturation than the native enzyme. At 90 °C, it retained 75% activity compared to the free enzyme. It was also more stable during storage at 4 °C: after 4 months the immobilized laccase retained 98% of initial activity. Immobilized C. unicolor laccase was active in 10–60% concentration of methanol, acetone, isopropanol or acetonitrile. The best enzymic activity was observed in 20% solution of acetonitrile in buffer.     

Maackiain in Cicer bijugum Rech. f. associated with resistance to Botrytis grey mould

Resistance in Cicer bijugum Rech. f. a wild relative of chickpea, to Botrytis grey mould (BGM), caused by Botrytis cinerea Pers., was shown to be associated with high concentrations of maackiain when compared to three susceptible species. The two BGM resistant accessions of C. bijugum contained between 200 and 300 μg maackiain g⁻¹ of foliage whereas the BGM susceptible species C. arietinum, C. echinospermum and C. reticulatum contained less than 70 μg g⁻¹. Furthermore, the concentration of maackiain increased to more than 400 μg g⁻¹ in the resistant wild species after being inoculated with the pathogen whereas no significant increase was recorded in the susceptible species. The germination of spores of Botrytis cinerea, treated with maackiain, was inhibited in a dose dependent manner; less than 10% of spores germinated when treated with 500 μg ml⁻¹. The data indicate that maackiain may be an important component in BGM resistance in the wild chickpea C. bijugum and that the resistance is enhanced in the presence of the pathogen.     

撕裂蜡孔菌在开放体系中对甲基橙染料的静态脱色研究

为了评价撕裂蜡孔菌处理偶氮染料的应用潜力,用性能稳定的甲基橙染料为材料,采用批次试验在开放性体系中研究了染料初始浓度、菌丝生物量、温度、pH等因素对该菌脱色能力的影响,运用菌丝体反接、染液光谱扫描、菌丝体显微观察等方法探讨了菌丝体脱色的可能机制,利用植物萌发试验进行了染料和脱色后溶液的毒性测试。结果表明,撕裂蜡孔菌在开放的静止体系中能够对甲基橙高效脱色,其最适脱色温度为35℃,最佳脱色pH值在6左右。菌丝对甲基橙的脱色表现在吸附和产酶降解两个方面,脱色过程中染料对菌丝体本身的影响较少。植物毒性分析显示撕裂蜡孔菌脱色48h后的产物对植物的毒性比甲基橙本身更强,若要彻底降解可能需要较长时间。本研究可为染料脱色工艺提供新的菌种。     

Antiectoparasitic activity of the gum resin, gum haggar, from the East African plant, Commiphora holtziana

The mechanism of ixodid tick (Acari: Ixodidae) repellency by gum haggar, a resin produced by Commiphora holtziana (Burseraceae), was investigated by evaluating activity against the cattle tick, Boophilus microplus. In an arena bioassay, a hexane extract of the resin of C. holtziana exhibited a repellent effect lasting up to 5 h. The hydrocarbon fraction of the resin extract was shown to account for the repellent activity, and was analysed by coupled gas chromatography–mass spectrometry (GC–MS). Major sesquiterpene hydrocarbons were tentatively identified as germacrene-D, δ-elemene and β-bourbonene. The identity and stereochemistry of the former compound was confirmed as the (+)-isomer by peak enhancement using enantioselective GC, whereas the latter 2 compounds, which are most likely degradation products of germacrene-type precursors, were identified through isolation by preparative gas chromatography followed by microprobe-NMR spectroscopy. GC comparison of gum haggar with another resin, C. myrrha, which was inactive in the tick bioassay, showed that the latter contained much lower levels of these hydrocarbons. To assess the suitability of the gum haggar resin as a general acarine repellent, further tests were made on a major acarine pest of European and US animal husbandry systems, the red poultry mite, Dermanyssus gallinae (Acari: Dermanyssidae). Gum haggar extract, and the isolated hydrocarbon fraction, showed strong repellent effects in an olfactometer assay, and again gum myrrh showed no effect. These findings provide a scientific basis for the observed anti-tick properties of gum haggar, and demonstrate the potential for its development as a general acarine repellent for use in animal husbandry systems.     

卵孢金孢藻对不同磷化合物的利用及生长响应

为了解水华蓝藻卵孢金孢藻对水体不同磷化合物的利用能力,试验以磷酸氢二钾为对照,通过室内模拟的方法探究不同磷基质条件下卵孢金孢藻的生长响应.结果表明: 卵孢金孢藻能直接利用三聚磷酸钠和十水焦磷酸钠,且对三聚磷酸钠有极高的利用率.15天后,三聚磷酸钠处理组的藻细胞生物量和叶绿素a浓度最高,分别达到(426.96±47.42) mg·L^-1和(1852.34±116.60) μg·L^-1.2-氨基乙基膦酸和五水β-甘油磷酸钠处理组生物量与对照无明显差异,可溶性无机磷变化特征响应了碱性磷酸酶活性大小,表明卵孢金孢藻能通过酶水解利用这两种有机磷.而整个培养过程中,草甘膦处理组可溶性无机磷浓度趋近于0 mg·L^-1,藻细胞生物量、比生长速率、叶绿素a浓度及光合活力均显著低于对照,表明草甘膦不仅不利于藻细胞摄取磷,还对其生长产生抑制作用.本研究结果为了解卵孢金孢藻向不同水生态系统的扩散机理提供了新的思路,对我国新型蓝藻水华的防治具有一定的理论参考价值.     

海南特有极小种群植物文昌锥的SCoT-PCR体系建立及有效引物评价

为了应用SCoT分子标记研究文昌锥种质资源,通过单因素试验和正交试验2种方法,分析DNA、引物、dNTPs、Taq DNA聚合酶这4种因素对文昌锥SCoT-PCR扩增结果的影响,优化建立文昌锥SCoT-PCR体系,并对文昌锥SCoT标记引物进行有效性评价。结果表明：各个因子对SCoT-PCR扩增影响大小依次为：DNA > dNTPs > 引物 > Taq DNA聚合酶;最优反应体系为：总体系为20 μL,DNA含量为2.5 ng,引物浓度为0.8 μmol·L^-1,dNTPs浓度为0.2 mmol·L^-1,Taq DNA聚合酶的含量1 U;经验证,该体系获得的扩增产物清晰、稳定;应用该体系筛选出15条多态性好且适合文昌锥扩增的引物,为今后利用SCoT分子标记技术对文昌锥及其他壳斗科植物进行相关研究提供技术支持。     

Lipase-catalyzed production of optically active (S)-flurbiprofen in aqueous phase reaction system containing chiral succinyl β-cyclodextrin

The lipase-catalyzed production of optically active (S)-flurbiprofen was carried out in a dispersion reaction-system induced by chiral succinyl β-cyclodextrin (suβ-CD). The optimal reaction conditions were 500 mM (R,S)-flurbiprofen ethyl ester ((R,S)-FEE), 600 units of Candida rugosa lipase per 1 mmol of (R,S)-FEE, and 1000 mM suβ-CD at 37 °C for 72 h. An extremely high enantiomeric excess of 0.98 and conversion yield of 0.48 were achieved in the dispersed aqueous phase reaction system containing chiral suβ-CD added as a dispenser and chiral selector. The inclusion complex formability of the immiscible substrate (S)- and (R)-form of FEE with suβ-CD was compared using a phase-solubility diagram, DSC, and ¹H NMR. (S)-Isomer formed a more stable and selective inclusion complex with chiral suβ-CD. It was hydrolyzed much more selectively by lipase from C. rugosa, due to the selective structural modification through inclusion complexation with chiral suβ-CD.     

Microwave assisted alkali-catalyzed transesterification of Pongamia pinnata seed oil for biodiesel production

In this study, microwave assisted transesterification of Pongamia pinnata seed oil was carried out for the production of biodiesel. The experiments were carried out using methanol and two alkali catalysts i.e., sodium hydroxide (NaOH) and potassium hydroxide (KOH). The experiments were carried out at 6:1 alcohol/oil molar ratio and 60 °C reaction temperature. The effect of catalyst concentration and reaction time on the yield and quality of biodiesel was studied. The result of the study suggested that 0.5% sodium hydroxide and 1.0% potassium hydroxide catalyst concentration were optimum for biodiesel production from P. pinnata oil under microwave heating. There was a significant reduction in reaction time for microwave induced transesterification as compared to conventional heating.     

Purification and characterization of an enantioselective carbonyl reductase from Candida viswanathii MTCC 5158

A highly enantioselective carbonyl reductase produced by a new yeast strain Candida viswanathii MTCC 5158, which was isolated using an acetophenone enriched medium, has been purified and characterized. The enzyme has been purified to near homogeneity using ammonium sulfate precipitation, ion exchange and gel filtration chromatography. The molecular properties of the carbonyl reductase suggested the native enzyme to be tetrameric, with an apparent molecular weight of 120 kDa, the monomer being about 29 kDa. Acetyl aryl ketones were found to be the preferred substrates for the enzyme and the best reaction was the enantioselective reduction of acetophenone. The enzyme yielded (S)-alcohol in preference to (R)-alcohol and utilized NADH, but not NADPH as the cofactor. The purified enzyme exhibited maximum enzyme activity at pH 7.0 and 60 °C. The enzyme retained about 80% of its activity after 7 h incubation at 25 °C in sodium phosphate buffer (50 mM, pH 7.0). The addition of reducing agents like dithiothreitol and β-mercaptoethanol enhanced the enzyme activity while organic solvents, detergents and chaotropic agents had deleterious effect on enzyme activity. Metal chelating agents like hydroxyquinoline and o-phenanthroline have significant effect on enzyme activity suggesting that the carbonyl reductase required the presence of a tightly bound metal ion for activity or stability. The maximum reaction rate (V_max) and apparent Michaelis–Menten constant (K_m) for acetophenone and NADH were 59.21 μmol/(min mg) protein and 0.153 mM and 82.64 μmol/(min mg) protein and 0.157 mM at a concentration range of 0.2–2 mM acetophenone (NADH fixed at 0.5 mM) and 0.1–0.5 mM NADH (acetophenone fixed at 2 mM), respectively.     

Cyperus rotundus suppresses AGE formation and protein oxidation in a model of fructose-mediated protein glycoxidation

Non-enzymatic glycation, as the chain reaction between reducing sugars and the free amino groups of proteins, has been shown to correlate with severity of diabetes and its complications. Cyperus rotundus (Cyperaceae) is used both as a food to promote health and as a drug to treat certain diseases. In this study, considering the antioxidative effects of C. rotundus, we examined whether C. rotundus also protects against protein oxidation and glycoxidation. The protein glycation inhibitory activity of hydroalcoholic extract of C. rotundus was evaluated in vitro using a model of fructose-mediated protein glycoxidation. The C. rotundus extract with glycation inhibitory activity also demonstrated antioxidant activity when a ferric reducing antioxidant power (FRAP) and Trolox equivalent antioxidant capacity (TEAC) assays as well as metal chelating activity were applied. Fructose (100 mM) increased fluorescence intensity of glycated bovine serum albumin (BSA) in terms of total AGEs during 14 days of exposure. Moreover, fructose caused more protein carbonyl (PCO) formation and also oxidized thiol groups more in glycated than in native BSA. The extract of C. rotundus at different concentrations (25–250 μg/ml) has significantly decreased the formation of AGEs in term of the fluorescence intensity of glycated BSA. Furthermore, we demonstrated the significant effect of C. rotundus extract on preventing oxidative protein damages including effect on PCO formation and thiol oxidation which are believed to form under the glycoxidation process. Our results highlight the protein glycation inhibitory and antioxidant activity of C. rotundus. These results might lead to the possibility of using the plant extract or its purified active components for targeting diabetic complications.     

Gene cloning of an NADPH-dependent menadione reductase from Candida macedoniensis, and its application to chiral alcohol production

The gene encoding an NADPH-dependent menadione reductase of Candida macedoniensis AKU4588 was cloned and sequenced. A 1035 bp nucleotide fragment (mer) was confirmed to be the gene encoding the enzyme based on the agreement of N-terminal and internal amino acid sequences. The mer encodes 345 amino acid residues, and the deduced amino acid sequence shows high similarity with those of hypothetical proteins from Debaryomyces, Candida and Saccharomyces, and ketoreductase from Zygosaccharomyces. It includes NADPH-binding motif GXXGXXA in its N-terminal region. These findings suggest that the enzyme belongs to the dihydroflavonol-4-reductase superfamily. An expression vector, pETMER, which contains the full length of the mer, was constructed. Escherichia coli cells harboring pETMER exhibits a 127-fold increase in specific menadione-reducing activity under the control of T7 promoter as compared with that of C. macedoniensis.

The asymmetric reduction of 4-chloro-3-oxobutanoate ethyl ester to (S)-4-chloro-3-hydroxybutanoate ethyl ester (CHBE) with E. coli cells, in which both the mer and the glucose dehydrogenase gene were co-expressed, as a catalyst was investigated. The (S)-CHBE formed amounted to 1680 mM (281 mg/ml), the molar yield being 92.2%. The optical purity of the product was 91.6% enantiomeric excess for the (S)-isomer. The calculated turnover number of NADP⁺ added to CHBE formed was 12,900 mol/mol.