近十年海洋来源木霉属真菌次生代谢产物研究进展

木霉（Trichoderma spp.）是一类自然界广泛存在于土壤中的真菌类群,属于半知菌门（Deuteromycotina）丝孢目（Moniliales）木霉属（Trichoderma sp.）。海洋中的木霉属真菌因其高盐、高压和缺氧的特殊环境,能够产生更多结构新颖、生物活性独特的次生代谢产物。木霉属真菌的次生代谢产物主要包括聚酮、肽类、萜类以及其他类型,这些化合物具有细胞毒、抗菌、酶抑制等多种生物活性。本文综述近十年来海洋来源木霉属真菌中分离的新次生代谢产物,并对这些新化合物的生物活性进行归纳总结。     

一株好氧反硝化菌的降酚特性及反硝化关键酶活性

为了研究菌株Diaphorobacter sp.PD-7在有氧条件下降解低浓度苯酚同时反硝化的特性,采用单因素试验,摇床转速180 r/min时,最佳降酚条件为200 mg/L乙酸钠和300 mg/L苯酚同时作为碳源,培养液初始p H为7.0,温度30℃,20 h内对初始浓度300 mg/L苯酚去除率达97.3%。培养液p H为中性时,硝酸盐还原酶(NAR)与亚硝酸盐还原酶(NIR)的最适反应温度分别为35℃、40℃。用米门方程对酶促反应动力学模拟,拟合曲线与实验测定值相关性良好,参数分别为Km=0.76μmol/L、vmax=42.02×10-3U/min和Km=1.49μmol/L、vmax=32.79×10-3U/min。推测NIR可能是该菌株反硝化过程中的限速酶,菌株PD-7主要通过细胞同化作用与反硝化作用脱氮,且该菌株有着完整的反硝化途径。     

两株细菌对偶氮染料活性艳红X—3B脱色的研究

从太原市纺织绒染厂废水处理站曝气池分离筛选到一株假单胞菌和一株黄单胞菌,探讨了这两株菌在不同碳源、不同ｐＨ和不同温度条件下对活性艳红Ｘ—３Ｂ的脱色效果。试验结果表明,假单胞菌和黄单胞菌分别以葡萄糖,果糖和无水乙酸钠为碳源时,脱色效果都较好,且黄单胞菌以可溶性淀粉作碳源时也有较高的脱色率。这两株菌脱色的最适ｐＨ均为８．０。假单胞菌脱色的最适温度为２５℃,黄单胞菌脱色的最适温度为３０℃。     

all 1691基因表达及铁元素对鱼腥藻生长的影响

为实现FurA的表达,探究铁元素对藻生长的影响,首先运用Primer5.0设计引物,克隆出鱼腥藻PCC7120染色体上all1691(furA)基因片段;构建含组氨酸融合表达标签和T7启动子标签的表达载体.IPTG诱导FurA蛋白表达.然后设计不同Fe3+浓度梯度培养基,利用SDS-PAGE检测高、中、低Fe3+浓度培养液的藻细胞总蛋白,考马斯亮蓝g-250法测定蛋白含量,并用Trizol法提取不同Fe3+浓度培养的藻细胞总RNA.结果表明,获得纯化的all 1691克隆片段,大小为456bp,pET-1691重组蛋白在1mmol/L的IPTG诱导下,于37℃下摇床培养15h得到成功表达;低浓度Fe3+促进藻生长,高浓度Fe3+抑制藻生长;Fe3+浓度为2.0mg/L时rRNA位置与亮度清晰可见,而缺铁培养的藻生长几乎停滞,转录和翻译水平都很低.     

短指软珊瑚Sinularia sp.内生真菌Ta31-2代谢产物研究(Ⅱ)

从短指软珊瑚Sinularia sp.内生真菌Ta31-2的代谢产物中分离得到10个化合物,利用波谱技术确定其结构分别为硬脂酸(1)、软脂酸(2)、肉豆蔻酸(3)、癸酸甲酯(4)、麦角甾醇(5)、fusanolide B(6)、丁二酸(7)、curvulalide(8)、modiolide A(9)和neoechinulin A(10)。其中,fusanolide B(6)为首次从海洋真菌中分离得到,curvulalide(8)和neoechinulin A(10)为首次从软珊瑚Sinularia sp.内生真菌中分离得到。     

球孢白僵菌产几丁质酶液态发酵研究

对球孢白僵菌Bb174产几丁质酶的液态发酵条件进行了试验研究。选择麸皮和蚕蛹粉作为培养基的基本成分。以不同比例的麸皮和蚕蛹粉进行试验,结果显示:麸皮和蚕蛹粉分别为5％,培养温度28℃,接种量10％,装液量10mL,起始pH6.0,添加0.3％NaNO_3对产儿丁质酶有利,最高酶活力为4664U/mL。     

环境因子对青海弧菌生长和发光的影响

该文采用了多因子正交试验与单因子试验相结合的方法研究了环境因子对青海弧菌性长和发光的影响,发现各因子对青海弧菌的生长和发光的影响程度是不同的。其中甘油对生长的影响最大,而对发光的影响最大的却是氮源。根据各因子的影响情况,本文提出了有利于青海弧菌生长和发光的致好的培养基配方及培养的温度条件,并试着对环境因子对青海弧菌生长和发光的影响机制作了一些讨论。     

红假单胞菌产类胡萝卜素条件的研究

通过对影响红假单菌类胡萝卜素的因素的全面研究,获得了胡萝卜素产量相对辊主贩培养条件,以乙酸钠为碳源,谷氨酸钠为氮源,酵母亮为生长因子,４ｍｇ／Ｌ,ＭｇＳＯ４,２ｍｇ／ＬｆＥ２（ｓｏ４）３,１５００ｌｘ光照度,３０℃,微好氧培养１２０ｈ。     

Effect of monoclonal antibody to bull sperm sp18 on murine fertilization and embryos development

Western blot analysis revealed that one IgG1 monoclonal antibody (mAb) to sp18 family membrane proteins (Mr. 14, 16 and 18 ku) of bovine sperm reacted faintly with protein bands of 14, 18, 22, 30 and 60 ku (reducing) in samples of mouse sperm. The mAb also reacted to protein of egg lysozyme. Using a laser confocal microscope, indirect immunofluorescence (IIP) showed that the sp18 antigens were present in the posterior head of murine sperm. In murine in vitro fertilization (IVF) and embryo development trails, a total of 426 oocytes from C57BL/6 and F1 hybrid strain (CD1 × C57BL/6 cross) of 12 female mice were used in 3 independent trails. After preincubating capacitated sperm with 182 μg/mL of sp18 mAb in the modified TYH IVF medium for 15-20 min, cumulus-oocyte-complexes were introduced. The fertilization rate in sp18 mAb groups was 77.1 %, which was not significantly (P > 0.05) different from the nonspecific mouse IgG (79.2%) and non-IgG (80.3 %) control groups. Fertilized oocytes had been continuously cultured in modifed CZB medium. 100%, 100% and 97.9% of 1-cell embryos developed to 2-cell stage in sp18 mAb, nonspecific mouse IgG and non-IgG group 30 h after the start of fertilization, respectively. In the nonspecific mouse IgG and non-IgG groups, 64.1 % and 64.3% of embryos developed to the 4-cell stage, respectively, but all developing eggs in sp18 mAb groups arrested development in vitro at 2-cell stage. After zonae of 2-cell blocked embryos were enzy-matically removed with 0.5% pronase, detection of sp18 antigens by IIF indicated that the fluorescence scattered on two embryonic cells. For embryos fertilized in vivo and co-cultured with 182 μg/mL sp18 mAb, the numbers of 1-cell embryos reaching the 2-cell and 4-cell stage were 95. 2% and 70. 5%, which were not significantly (P>0.05) different from the control group (92.9% and 77.9%). These results indicate that the sp18 antigens on posterior head of mouse sperm were incorporated into the egg plasma membrane during fertilization, and played an active role in development of murine preimplanta-tion embryo.     

脉孢霉纤溶酶的酶学性质研究

研究了脉孢霉纤溶酶的部分酶学性质,确定该酶由双亚基组成,亚基分子量分别为30000Da、15500Da,等电点为7.9±0.2,最适作用pH值为7.4,最适作用温度为50℃,在pH6.6～9和32～42℃时较稳定。