栉孔扇贝血淋巴中酯酶活性及其性质

采用紫外分光光度法测定栉孔扇贝（Chlamysfarreri）血淋巴中酯酶的活性，并研究底物、抑制剂、激活剂、温度、pH对酯酶活力的影响及酶的热稳定性。结果表明，栉孔扇贝血清与血细胞中均存在酯酶，且血细胞中的酶活力高于血清。血清与血细胞中酯酶的最适作用底物有所不同，血清中为α-醋酸萘酯，而血细胞中为α-丁酸萘酯。当以α-醋酸萘酯为底物时，血清中酯酶的最适温度为45℃，最适pH值为7．5；以α-丁酸萘酯为底物时，血清中酯酶的最适温度为40℃，最适pH值为8．0。以α-醋酸萘酯及α-丁酸萘酯为底物分别测定酶的热稳定时，酶活力变化相似，在20℃时活力最高，而后缓慢下降。栉孔扇贝血淋巴中酯酶属中温型中性酶类，低温下较稳定，底物专一性不强，血清中的酯酶可能来自血细胞，该酶在贝类的免疫防御中町发挥重要的作用。     

Intestinal phytase I: Detection and preliminary characterization of activity in the intestinal brush border membrane of hybrid striped bass Morone saxatilis × M. chrysops

While numerous studies have evaluated intestinal phytase activity in terrestrial monogastric species, little work has been conducted investigating the enzyme's activity in finfish. This experiment was conducted to determine if hybrid striped bass Morone saxatilis × M. chrysops possess intestinal phytase activity, and to perform a preliminary characterization of this activity in terms of pH, metal ion dependence and substrate kinetics. Hybrid bass do exhibit low levels of intestinal phytase activity. The activity has an acidic pH optimum between 3.5 and 4.5. Low concentrations of Mn²⁺ slightly enhance this activity. The divalent cations Mg²⁺ and Zn²⁺ were shown to either have no effect on the enzyme activity or to be inhibitory, particularly at high concentrations. The present results indicate that intestinal phytase activity in hybrid bass does follow Michaelis-Menten kinetics, with estimated K_m and V_max values of 2.5 mM phytate and 4.8 units mg⁻¹ protein, respectively. The physiologic significance of the enzyme's presence in hybrid striped bass is questionable due to the low activity as compared to terrestrial monogastrics, the acidic pH optimum and the low affinity the enzyme appears to have for phytate. These observations also raise the question of whether the intestinal phytase activity detected in hybrid bass is merely a manifestation of non-specific acid phosphatase activity. Further investigation is needed to determine the practical significance of phytase activity in hybrid bass in terms of phytate phosphorus utilization. Additional experiments should also be conducted which evaluate intestinal phytase activity in other important aquacultured finfish species. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effect of shrimp head protein hydrolysates on the state of water and denaturation of fish myofibrils during dehydration

ABSTRACT:   To utilize fisheries waste products as food materials with functional properties, shrimp head protein hydrolysates (SHPH) from three species of shrimp, that is, Northern pink shrimp ( Pandalus eous ), Endeavour shrimp ( Metapenaeus endeavouri ) and Black tiger shrimp ( Penaeus monodon ), were produced by enzymatic hydrolysis using endopeptidase derived from Bacillus subtilis and exopeptidase derived from Aspergillus oryzae at a level of 0.1% (w/w). SHPH were rich in protein (90–91%) and amino acids (71–84%) but little fat (0.01–0.02%). The average molecular weight of SHPH was 300–1400. The effect of 5% SHPH (dry basis) addition on the state of water and denaturation of lizard fish myofibrils (Mf) during the dehydration process was evaluated by the desorption isotherm and the Ca-ATPase activity, and compared with the effect of sodium glutamate (Na-Glu). SHPH decreased the water activity and the Ca-ATPase inactivation, and increased monolayer sorbed water and multilayer sorbed water of Mf, although these effects of SHPH were smaller than those of Na-Glu. These findings suggest that the SHPH suppressed dehydration-induced denaturation of myofibrillar protein by stabilizing the hydrated water surrounding myofibrils.     

巴西橡胶树橡胶粒子中2个含RicinB_Lectin_2结构域蛋白质研究初报

采用不同提取液,从洗涤过的巴西橡胶树橡胶粒子中提取蛋白质,并对其进行电泳分离与质谱鉴定。分离鉴定了2个含RicinB_Lectin_2结构域的糖基识别蛋白质,分别命名为HbRBLLP-1和HbRBLLP-2。这2个蛋白质可能是橡胶粒子膜蛋白质或是与橡胶粒子有较强的相互作用的蛋白质。对HbRBLLP-1及HbRBLLP-2进行生物信息学分析,结果表明它们的RicinB_Lectin_2结构域均仅有1个Q-x-W模体,而非蓖麻毒素B链所具有的(Q-x-W)3模体。     

GST-MtDef4融合蛋白的原核表达及体外抑菌活性的检测

为研究苜蓿(Medicago truncatula)防卫素基因MtDef4的抑菌功能,以含有该基因片段的质粒pAND-MtDef4为模板,扩增出相应的目的基因,测序正确后将MtDef4基因片段连接到带有GST标签的原核表达载体pGEX6P-1中,转化大肠杆菌TransB(DE3),经IPTG诱导,Glutathione Sepharose 4B亲和纯化,通过SDS-PAGE电泳及Western blot确定融合蛋白的表达。体外抑菌试验表明,MtDef4能够抑制病原真菌Corynespora cassiicola而非Collectotrichum gloeosporioides的生长。本研究结果可为进一步培育转基因橡胶抗病品种奠定基础。     

NR/ENR/SiO_2复合材料的阻尼性能研究

采用机械混炼的方法制备不同ENR含量的NR/ENR/SiO2复合材料,使用动态热机械分析仪(DMA)和橡胶加工分析仪(RPA)对NR/ENR/SiO2复合材料的动态力学性能和阻尼性能进行分析。结果表明:ENR对NR/ENR/SiO2复合材料的阻尼性能具有显著影响。动态热机械分析测试结果表明,随着ENR用量的增多,NR/ENR/SiO2复合材料在使用温度范围内的损耗因子积分面积增大,阻尼性能提高。橡胶加工分析频率扫描和应变扫描测试结果表明,在频率小于10 HZ不同应变时,加入了ENR的NR/ENR/SiO2复合材料的阻尼因子均高于NR/SiO2复合材料。扫描电镜分析结果证实,ENR的加入减少了SiO2的自聚,改善了填料在橡胶基体中的分散,从而使NR/ENR/SiO2复合材料力学性能得到提高。     

高速逆流色谱法制备分离库拉索芦荟中的2’-对香豆酰芦荟宁

采用高速逆流色谱法分离纯化库拉索芦荟中的2’-对香豆酰芦荟宁。以正己烷-乙酸乙酯-丙酮-水(1∶5∶1∶5,V/V/V/V)为溶剂体系,上相为固定相,下相为流动相,线圈转速为860 r/min,流速为2 mL/min,紫外检测波长为300 nm,一次进样85 min内从487.5 mg库拉索芦荟丙酮提取物中分离得到19.5 mg单体组分,经高效液相色谱分析,测定纯度为95.6%(峰面积百分比),通过质谱、红外、紫外和核磁技术进行结构鉴定,确定组分为2’-对香豆酰芦荟宁。该法具有操作简便、速度快、样品制备量大、节省溶剂及回收率高等优点,可为芦荟宁类化合物的分离提供高效、稳定及可靠的方法。     

国家自然科学基金项目（J1210056）

为探讨硫化氢(H2S)作为潜在的新型氮肥增效剂的可能性,通过水培实验,以扬麦16为材料,以NaHS为H2S供体(浓度0.01mmol·L-1),研究了外源低浓度H2S对低氮(2.5 mmol·L-1)、中氮(7.5)和高氮(15 mmol·L-1)条件下小麦幼苗光合作用、硝态氮吸收和同化的影响.结果表明,外源低浓度H2S促进了小麦幼苗的光合作用及对硝态氮的吸收,使低氮条件下植株干物质重增加15.5％,显著提高叶片可溶性蛋白、总氮及叶绿素含量.外源H2S处理后,低氮和中氮水平下叶片硝酸还原酶(NR)、谷氨酰胺合成酶(GS)以及谷氨酸脱氢酶(GDH)活性有所增加,尤其是在低氮水平下增加显著,但在高氮水平下NR与GS活性分别下降22％和13％,GDH活性与对照无明显差异.以上结果说明外源低浓度H2S可提高小麦幼苗对低氮的适应性,促进其生长及对氮素吸收与同化.     

Aaptamine Derivatives with Antifungal and Anti-HIV-1 Activities from the South China Sea Sponge Aaptos aaptos

Five new alkaloids of aaptamine family, compounds (1–5) and three known derivatives (6–8), have been isolated from the South China Sea sponge Aaptos aaptos. The structures of all compounds were unambiguously elucidated by spectroscopic analyses, as well as by comparison with the literature data. Compounds 1–2 are characterized with triazapyrene lactam skeleton, whereas compounds 4–5 share an imidazole-fused aaptamine moiety. These compounds were evaluated in antifungal and anti-HIV-1 assays. Compounds 3, 7, and 8 showed antifungal activity against six fungi, with MIC values in the range of 4 to 64 μg/mL. Compounds 7–8 exhibited anti-HIV-1 activity, with inhibitory rates of 88.0% and 72.3%, respectively, at a concentration of 10 μM.     

Anti-Mycobacterial Nucleoside Antibiotics from a Marine-Derived Streptomyces sp. TPU1236A

Five new nucleoside antibiotics, named streptcytosines A–E (1–5), and six known compounds, de-amosaminyl-cytosamine (6), plicacetin (7), bamicetin (8), amicetin (9), collismycin B (10), and SF2738 C (11), were isolated from a culture broth of Streptomyces sp. TPU1236A collected in Okinawa, Japan. The structures of new compounds were elucidated on the basis of their spectroscopic data (HRFABMS, IR, UV, and 2D NMR experiments including ¹H-¹H COSY, HMQC, HMBC, and NOESY spectra). Streptcytosine A (1) belonged to the amicetin group antibiotics, and streptcytosines B–E (2–5) were derivatives of de-amosaminyl-cytosamine (6), 2,3,6-trideoxyglucopyranosyl cytosine. Compound 1 inhibited the growth of Mycobacterium smegmatis (MIC = 32 µg/mL), while compounds 2–5 were not active at 50 µg/disc. Bamicetin (8) and amicetin (9) showed the MICs of 16 and 8 µg/mL, respectively.