Study of two-stage processes for the microbial production of 1,3-propanediol from glucose

The microbial production of 1,3-propanediol (1,3-PD) from glucose was studied in a two-stage fermentation process on a laboratory scale. In the first stage, glucose was converted to glycerol either by the osmotolerant yeast Pichia farinosa or by a recombinant Escherichia coli strain. In the second stage, glycerol in the broth from the first stage was converted to 1,3-PD by Klebsiella pneumoniae. The culture broth from P. farinosa was shown to contain toxic metabolites that strongly impair the growth of K. pneumoniae and the formation of 1,3-PD. Recombinant E. coli is more suitable than P. farinosa for producing glycerol in the first stage. The fermentation pattern from glycerol can be significantly altered by the presence of acetate, leading to a significant reduction of PD yield in the second stage. However, in the recombinant E. coli culture acetate formation can be prevented by fed-batch cultivation under limiting glucose supply, resulting in an effective production of 1,3-PD in the second stage with a productivity of 2.0 g l(-1) h(-1) and a high yield (0.53 g/g) close to that of glycerol fermentation in a synthetic medium. The overall 1,3-PD yield from glucose in the two stage-process with E. coli and K. pneumoniae reached 0.17 g/g.     

Pollen and resource limitations to lifetime seed production in a wild population of the endangered plant Disanthus cercidifolius var. longipes H. T. Chang (Hamamelidaceae)

Disanthus cercidifolius Maxim. var. longipes H. T. Chang, a plant species that only occurs in a few counties in Hunan, Jiangxi and Zhejiang Provinces and with a relatively small number of individuals, is ranked as a second Class endangered species for conservation in China. We have studied the effect of pollen and resources available to female reproduction, and the reproductive mechanism of “excess flowers with low fruit set” in Disanthus cercidifolius Maxim. var. longipes H. T. Chang was discussed. Results are as follows Pollen from different sources has significant effects on fruit set and seed set of Disanthus cercidifolius Maxim. var. longipes H. T. Chang. The pollen source rather than pollen numbers significantly affected reproduction of this species. In wild populations, producing one fruit needs about 54.8 flowers, and one satiation seed needs about 6.60 flowers or 83.19 ovules. After fertilizing, which was propitious to flower development, the abortion rate of flower buds was decreasing, but the flowering rate was increasing. The fruit set and seed set was also significantly increasing, while abortion rate of fruit was significantly decreasing. With the increasing percentages of cutting leaves, the fruit set decreased, but the abortion rate of fruit shows no significant differentiation among treatments. After cutting branches that were puny, broken and insectin-fested branches, the flower number seemed to be decreasing, but the fruit set and seed set all increased significantly. After removing some flowers, the fruit set was calculated with respect to the number of flowers remaining after the treatment increased with increasing of percentages of flower removal, whereas fruit set calculated with respect to the initial number of flowers remained constant, and the mean weights of per fruit and per seed all decreased significantly. Sufficient spatial or temporal heterogeneities in nutrient levels might allow limitation of seed set by resources and pollen in a natural population, while supplying resources may indirectly affect pollination by increasing attraction of the flowers to pollinators. There were very low fruit and seed sets in natural populations of Disanthus cercidifolius Maxim. var. longipes H. T. Chang. Different factors may have interacted to effect a low fruit set. A joint adoption of the “selection abortion hypothesis”, “ovary reserve hypothesis” and “male function hypothesis” seems to be the most likely explanation for the reproductive strategy of “excess flowers with few fruit sets” in Disanthus cercidifolius Maxim. var. longipes H. T. Chang.     

新疆地震断裂带次生植物根际土壤微生物碳源利用

利用BIOLOG技术研究了新疆富蕴地震断裂带6种次生植物对根际土壤微生物碳源利用的影响.结果表明:多数植物根际土壤养分显著高于对照.6种根际土壤的平均颜色变化率差异显著,且均高于对照.次生植物不影响根际土壤微生物碳源利用的丰富度,但改变了其优势度和均匀度.不同处理根际土壤微生物碳源利用存在差异,主要体现在对糖类、氨基酸类、羧酸类的利用上.断裂带上次生植物的出现使土壤微生物利用碳源的类型由酚类向糖类、羧酸类转变.土壤速效钾含量与土壤微生物对聚合物(r=-0.84)、胺类(r=-0.83)的利用呈负相关.新疆地震断裂带的次生植物能显著增强土壤微生物的碳源利用能力,改变碳源利用类型.密刺蔷薇和蓍在提高土壤微生物碳源利用能力和改善土壤养分方面效果最佳.     

三门湾大型底栖动物时空分布及其与环境因子的关系

2006年11月、2007年1月、4月和8月在三门湾18个采样点对大型底栖动物进行调查,分析了其时空分布及其与环境因子的关系.结果表明:调查共采集到大型底栖动物124种,其中多毛类44种、软体动物34种、甲壳动物22种、棘皮动物11种、其他类动物13种;多毛类和软体动物种数占总种数的62.9％,二者构成了三门湾大型底栖动物的主要类群.双鳃内卷齿蚕、小头虫和不倒翁虫是春季三门湾大型底栖动物的优势种;不倒翁虫、双鳃内卷齿蚕和海稚虫为夏季的优势种;不倒翁虫、小头虫、双鳃内卷齿蚕和白沙箸为秋季的优势种;双鳃内卷齿蚕、不倒翁虫、小头虫和海稚虫为冬季的优势种.三门湾大型底栖动物年均生物量为17.36 g·m-2,年均栖息密度为72 ind·m-2.不同季节大型底栖动物的平均生物量和平均栖息密度存在显著性差异.大型底栖动物群落平均Shannon多样性指数在1.53 ～ 1.89,平均Margalef物种丰富度指数在2.25 ～2.96,平均均匀度指数在0.83 ～0.94,3个指数在不同季节间均存在显著性差异.经典范对应分析,影响三门湾大型底栖动物群落的主要环境因子包括海水的温度、盐度、溶解性无机氮以及表层沉积物中的有机质、总氮和总磷等,环境变量可以较好地解释主要类群的变化.     

从一名国内感染的艾滋病人分离人免疫缺陷病毒（HIV）

从一名国内感染的艾滋病人采血,分离其外周血单核细胞（ＰＭＣｓ）。首先与正常的ＰＭＣｓ共培养,４周后检测其ＨＩＶ－１ｐ２４抗原（ＥＬＩＳＡ）达到峰值。用此时的细胞及其上清分别感染Ｊｕｒｋａｔ－ｔａｔ、ＣＥＭ、ＭＴ４细胞,可很快地在这三株细胞中检测到ＨＩＶ生长,ＨＩＶ在Ｊｕｒｋａｔ－ｔａｔ细胞中生长情况最好,同时用病人血清直接感染Ｊｕｒｋａｔ－ｔａｔ和ＭＴ４细胞,４周后检测其细胞上清ＨＩＶ－１ｐ２４抗原（ＥＬＩＳＡ法）为阳性,但ＯＤ值很低（约为ＰＭＣ共培养组的一半）。用病人的少量全血与正常ＰＭＣｓ共培养,得到的结果与分离病人ＰＭＣｓ法相近。应用间接免疫荧光法（ＩＦＡ）、免疫酶法（ＩＥＡ）、蛋白印迹法及ＨＩＶ－１ＰＯｌ基因和Ｅｎｖ基因特异引物的聚合酶链反应（ＰＣＲ）等证实为ＨＩＶ－１病毒。分离的ＨＩＶ在Ｊｕｒｋａｔ－ｔａｔ细胞中连续传代,细胞被感染后２－３天即出现以大量融合细胞为主的细胞病变,感染后７－１０天细胞几乎全部死亡。病毒在连续传代过程中的生长特征及致细胞病变特征不变。此病毒命名为ＣＡ－２毒株。     

大鼠主要脏器中微量元素含量研究

用原子吸收光谱法测定了铜、铁、锌、锰、铬、镁、铅、钾在SPF大鼠心、肝、肾、肺、脾中的含量,发现铜、锌、铁、锰、在肝脏中浓度最高（P＜0．001）;铬、钾在肺脏中浓度最高（P＜0．001）;镁、铅在各脏器中浓度无显著性差别（P＞0．05）。     

Efficient mucosal vaccination mediated by the neonatal Fc receptor

Almost all infectious diseases are initiated at mucosal surfaces, yet intramuscular or subcutaneous vaccination usually provides only minimal protection at sites of infection owing to suboptimal activation of the mucosal immune system. The neonatal Fc receptor (FcRn) mediates the transport of IgG across polarized epithelial cells lining mucosal surfaces. We mimicked this process by fusing a model antigen, herpes simplex virus type-2 (HSV-2) glycoprotein gD, to an IgG Fc fragment. Intranasal immunization, together with the adjuvant CpG, completely protected wild-type, but not FcRn knockout, mice after intravaginal challenge with virulent HSV-2 186. This immunization strategy induced efficient mucosal and systemic antibody, B- and T-cell immune responses, with stable protection for at least 6 months after vaccination in most of the immunized animals. The FcRn-IgG transcellular transport pathway may provide a general delivery route for subunit vaccines against many mucosal pathogens.     

Proteomic analysis of silk gland programmed cell death during metamorphosis of the silkworm Bombyx mori

The silk gland of the silkworm Bombyx mori undergoes programmed cell death (PCD) during pupal metamorphosis. On the basis of their morphological changes and the occurrence of a DNA ladder, the tissue cells were categorized into three groups: intact, committed, and dying. To identify the proteins involved in this process, we conducted a comparative proteomic analysis. Protein expression changes among the three different cell types were examined by two-dimensional gel electrophoresis. Among approximately 1000 reproducibly detected protein spots on each gel, 43 were down-regulated and 34 were up-regulated in PCD process. Mass spectrometry identified 17 differentially expressed proteins, including some well-studied proteins as well as some novel PCD related proteins, such as caspases, proteasome subunit, elongation factor, heat shock protein, and hypothetical proteins. Our results suggest that these proteins may participate in the silk gland PCD process of B. mori and, thus, provide new insights for this mechanism.