Purification and characterization of a 630 kDa bacterial killing metalloprotease (KilC) isolated from plaice Pleuronectes platessa (L.), epidermal mucus

Antibacterial chemicals in the mucus of fish such as lysozyme, lectins, peptides and proteases provide an efficient first line of defence against pathogens. This study shows that there are at least three antibacterial proteins in plaice skin mucus in addition to lysozyme. One of these proteins is responsible for approximately 74% of the antibacterial activity and is a 630 kDa protease complex designated KilC (bacterial killing metalloprotease C). Purified KilC kills the bacteria Staphylococcus aureus, Escherichia coli, Bacillus subtilis and Pseudomonas aeruginosa efficiently. The protease activity of KilC is dependent upon the divalent cation Mg(2+) and shows pH dual optima of 5.0 and 8.0. The enzyme has a temperature optimum of 25 degrees C and is made up of at least five different sized peptides. Studies with protease inhibitors show that the catalytic site of KilC may be cysteine- or serine protease-like. KilC may kill bacterial cells by acting directly upon the bacteria or by producing low molecular weight bioactive compounds such as peptides.     

Seminal plasma proteins of Atlantic halibut (Hippoglossus hippoglossus L.)

Protein content and properties in the seminal plasma of Atlantic halibut (Hippoglossus hippoglossus) were assayed using spectrophotometric and electrophoretic methods. The protein concentration ranged from 6.4 ± 3.1 to 19.4 ± 3.4 mg ml⁻¹ and anti-proteolytic activity from 585.2 ± 104.6 to 2912.4 ± 367.4 U l^−l. A high correlation between anti-proteolytic activity and protein concentration (r = 0.95), and between sperm concentration and osmolality was found (r = 0.92). There was a significant decrease in anti-proteolytic activity from the first to the second sampling, but not in protein concentration. Anti-proteolytic activity and protein concentration were significantly affected by variations in individual males. Electrophoresis revealed four anti-proteolytic bands and individual differences in bands of proteolytic activity, which were subsequently characterized as metalloproteases and serine proteases.     

Interactions between fishing strategies of Nephrops trawlers in the Bay of Biscay and Norway lobster diel activity patterns

Abstract  Norway lobster, Nephrops norvegicus (L.), in the Bay of Biscay exhibited diel activity patterns with more individuals outside their burrows at dawn and dusk, increasing catchability at these times. Data from an on board observer programme on Nephrops trawlers between 2002 and 2005 were used to assess variability in catchability in commercial catches. Catch numbers per haul varied spatially and between months, but no signal for diel variations was found. Fishing strategies developed by the Nephrops trawlers had several components. On a seasonal level, they started around sunrise. On a haul level, haul duration decreased from haul to haul, with the longest hauls taking place at the time of the highest catchability. By-catch of hake, Merluccius merluccius (L.), increased more than proportionally with haul duration.     

小兴安岭谷地云冷杉林土壤酶活性的异质性

利用传统统计学与地统计学相结合的方法对小兴安岭谷地不同死亡程度云冷杉林表层土壤（0~20cm）酶活性空间异质性和格局进行研究。结果表明：①土壤脲酶变异函数曲线的理论模型符合线性模型,土壤转化酶、碱性磷酸酶和过氧化氢酶的理论模型符合指数模型或球状模型。②土壤酶活性各项指标的空间变异主要是由结构性因素引起,土壤碱性磷酸酶活性和过氧化氢酶活性指标的空间自相关程度均属较强以上（空间结构比均在75%以上）,土壤转化酶为中等（空间结构比均在25%以上）,土壤脲酶在较弱到中等之间变化。③云冷杉林表层土壤过氧化氢酶的自相关范围最大（0.58~3.08 m）,转化酶和碱性磷酸酶分别为1.41~3.98 m和0.91~4.29 m,而土壤脲酶自相关范围最小（15.426~16.673 m）。④土壤转化酶、碱性磷酸酶和过氧化氢酶活性的空间格局明显,土壤转化酶、碱性磷酸酶、过氧化氢酶和土壤脲酶的分数维分别为1.930~1.990,1.910~1.986,1.923~1.997和1.936~1.970。     

广东陈香茶后发酵过程中主要微生物种群和酶类活性变化的研究

以岭头单丛茶树品种晒青毛茶为材料进行后发酵加工陈香茶,重点跟踪研究后发酵过程不同阶段的微生物种群、主要生化成分及主导酶类的变化并进行鉴定和分析。结果表明,在陈香茶后发酵过程中,微生物以产黄青霉和黑曲霉为主。茶多酚、茶黄素、茶红素、游离氨基酸等茶叶品质成分含量均呈下降趋势,而茶褐素逐步增加;果胶酶、纤维素酶等促进茶叶品质形成的酶类在后发酵一翻（4 d）时活力到达最高峰,多酚氧化酶在整个过程中活性较低。     

HXKs基因在香蕉果实后熟过程中功能初探

香蕉是世界主要水果之一,对乙烯非常敏感,属于呼吸跃变型果实。目前,香蕉果实后熟机理还未完全探明,后熟过程中己糖激酶(HXK)与乙烯的关系还不清楚。通过BLAST比对从香蕉基因组数据库中发现HXK基因家族的11个成员,经克隆获得这些基因的序列,并研究HXKs基因在香蕉果实自然后熟过程中的转录表达谱,重点研究乙烯利、甘露糖、NAG和1-MCP对HXKs基因表达、HXK酶活和内源乙烯生物合成的影响。结果表明:在香蕉果实后熟过程中HXKs基因呈现差异性表达,乙烯利上调大多数HXKs基因的表达和HXK酶活,1-MCP的作用正好相反,这表明外源乙烯正作用于HXK。另一方面,甘露糖加快内源乙烯生物合成,NAG却推迟内源乙烯生物合成,这说明HXK正作用于内源乙烯生物合成。所以,在香蕉果实后熟过程中乙烯和HXK之间存在相互促进的关系。这将为进一步阐明香蕉果实后熟机制提供理论依据,也将为香蕉果实保鲜新技术的挖掘提供新思路。     

Resistance Evaluation to Sheath Blight in Transgenic Rice Lines

Resistance of forty-one homozygous rice lines transformed with chitinase gene (RC24) and β-1,3 -glucanse gene β-1,3-Glu) to sheath blight was analyzed by inoculation. Among different lines, the resistance had significant differences according to the result by cluster analysis. The lines could be categorized into resistant, moderately resistant, moderately susceptible and susceptible types, while 92.1% of which belonging to moderately resistant or moderately susceptible type. For different resistant or susceptible lines, the resistance to rice sheath blight was remarkable correlated with the chitinase activity of transgenic lines except resistant type lines, in which enzyme activity coded by target gene was lower than moderately resistant type. The chitinase activity of transgenic lines tested at different time after inoculation or different organs of the same plant was uniform, which suggested that the expression of chitinase gene was constitutive in nature. Check varieties chitinase activity would change at different time after inoculation and reach a peak at sometime, but it had no difference at various pans of the same plant.     

整株套罩对桂味荔枝果实贮藏特性的影响

以套罩及对照桂味荔枝为材料,统计贮藏期果实病果率,测定果实糖、酸、Vc含量,抗氧化酶SOD、POD及褐化相关酶PPO的活性,以此分析整株套罩对荔枝贮藏性的影响。结果显示：4 ℃条件下,套罩桂味贮藏两周无病果,且果肉可溶性糖在4.0～6.0 mg/g之间,可滴定酸在2.5～3.0 mg/g之间,品质较好。清除氧化自由基方面,贮藏两周后,套罩桂味果皮MDA含量显著低于对照,果皮Vc含量、POD活性与对照无显著性差异,但其SOD活性显著高于对照;酶促褐化方面,套罩桂味果皮PPO活性与对照亦无显著差异。与果皮不同,套罩桂味果肉MDA含量显著高于对照,但果肉Vc含量、SOD、POD以及PPO活性在二者间无显著差异。试验结果表明,果皮抗氧化能力是决定桂味荔枝耐贮性的主要因素,套罩桂味依赖较高的果皮SOD活性维持其清除自由基的能力,并以此增强果实贮藏性。     

发酵诺尼果种子中有效成分及其抗氧化性

为了有效利用已发酵诺尼果(Noni)种子,探讨提取发酵诺尼种子中有效成分的方法,进一步研究发酵诺尼果种子有效成分的抗氧化性。笔者分别用水蒸气蒸馏和有机溶剂95%乙醇萃取,再对2种提取方法分别获得的发酵诺尼果提取物进行气相色谱质谱(GC-MS)分析;采用A320电子顺磁共振波谱仪对乙醇萃取获得的发酵诺尼果种子精油的抗氧化性进行分析。结果表明,发酵诺尼果种子提取物中的有效成分绝大多数为酯类物质,此外还含有酸类和醇类物质。95%乙醇萃取物经GC-MS分析,首次在发酵诺尼果种子提取物中发现了角鲨烯,其相对含量为12.95%。电子顺磁共振法可简便、快速检测发酵诺尼果种子精油的抗氧化性,乙醇萃取发酵诺尼果种子精油不同层DPPH清除能力表现不同,在提取物相同体积时,样品清除自由基的反应速度很快,其中乙醇萃取发酵诺尼果种子精油下层在2 min能够完全清除自由基,而精油上层在20min不能完全清除自由基。说明其精油下层的抗氧化活性高于上层。笔者认为,已发酵诺尼果种子含有开发价值的活性物质,且该活性物质具有抗氧化性。     

磷水平对杂交水稻及其亲本根系酸性磷酸酶活性的影响

为了解优良亲本和杂交组合的磷营养遗传特性,以7份亲本及其4个组合为材料,采用水培试验研究了不同磷水平对水稻亲本及其杂交组合根系酸性磷酸酶(APase)活性的影响。在低磷条件下,磷低效型保持系材料Ⅱ-32B的APase活性较对照增加不显著,而磷高效型保持系材料D62B和D83B则通过显著提高根系APase活性增强了对磷胁迫环境的适应性。磷高效型恢复系材料R892和R527在分蘖期和孕穗期的APase活性均较对照显著提高,而磷低效型恢复系材料R549和R781除在分蘖期APase活性增加明显外,在孕穗期和灌浆期APase活性与对照差异不显著。不同亲本配制的杂交稻在低磷水平下,根系APase活性增加的幅度有所不同。磷低效型杂交组合Ⅱ优549分蘖期、孕穗期和灌浆期根系的APase活性在不同供磷水平下差异不显著;磷高效型杂交组合D83A/R527在低磷水平下3个时期APase活性均明显提高。由磷低效型保持系材料Ⅱ-32B与磷低效型恢复系材料R549配制的Ⅱ优549,根系APase活性受低磷胁迫增幅不大;磷高效型保持系材料D83B与磷高效型恢复系材料R527配制的D83A/R527,根系APase活性在低磷水平下上升显著;磷低效型保持系材料Ⅱ-32B与磷高效型恢复系材料R892配制的Ⅱ优892,以及磷高效型保持系材料D62B与磷低效型恢复系材料R781配制的D62A/R781,受低磷胁迫时根系APase活性上升幅度介于磷低效组合Ⅱ优549和磷高效组合D83A/R527之间。