Enhanced Biotransformation of 2-Phenylethanol with Ethanol Oxidation in a Solid–Liquid Two-Phase System by Active Dry Yeast

2-Phenylethanol (2-PE) can be produced from l-phenylalanine (l-Phe) with the oxidation degradation of ethanol by active dry yeast. In this study, the catalysis effect of ethanol on biotransforming l-Phe into 2-PE by yeast was evaluated and optimized. The results indicated that increasing ethanol concentration was beneficial for enhancing 2-PE concentration but lowered the 2-PE productivity. Initial ethanol concentration above 25 g/l could strongly inhibit the 2-PE production. To obtain 2-PE with desirable concentrations with an economical operation mode, three fed-batch biotransformation operation methods using ethanol or/and glucose were carried out in a solid–liquid two-phase system. When using ethanol alone with the initial concentration of 10 g/l, the total concentration and overall productivity of 2-PE were 7.6 g/l and 0.065 g l⁻¹ h⁻¹, respectively. Furthermore, an experiment with controlled glucose solely (higher than 2 g/l) was finished. In this case, phenylacetaldehyde (PA) was detected along with ethanol accumulation, suggesting that reaction of PA → 2-PE in Ehrlich pathway was inhibited. To further enhance 2-PE production by using glucose only, a novel operation strategy to simultaneously control rates of glucose glycolysis and ethanol oxidative degradation with the aid of ISPR techniques was developed. With this strategy, 2-PE concentration and yield based on glucose consumption reached a higher level of 14.8 g/l and 0.12 g-PE/g-glucose, respectively, and these are the highest values reported up to date with the fed-batch biotransformation operation mode.     

溶瘤腺病毒ZD55对类肝癌干细胞的抑制效应

溶瘤腺病毒能够靶向和杀死癌症干细胞,被认为是一种很有前景的抗癌药物.已有研究表明,溶瘤腺病毒ZD55能够靶向肝癌,并且表现出明显的细胞毒性效应.然而,其对肝癌干细胞是否具有同样地杀伤效力仍需进一步探讨.利用悬浮培养富集类肝癌干细胞,并验证其肝癌干细胞的特征.进一步通过MTT、结晶紫染色、Hoechst染色、Western blot和流式细胞术等检测ZD55对类肝癌干细胞的细胞存活率、凋亡诱导和病理效应等.结果发现,悬浮培养的类肝癌干细胞具有自我更新和分化能力、高表达干细胞相关转录因子(如NANOG和OCT4)、处于静息状态和具有耐药性等特性,溶瘤腺病毒处理后表现出明显的细胞毒性效应和杀伤特性,类肝癌干细胞的最低生存率仅为26.7%.ZD55能够非常明显地诱导类肝癌干细胞凋亡,其凋亡率最高达到60%.因此,ZD55可能会成为靶向肝癌干细胞的一种很有前景的治疗药物,对肝癌的临床治疗具有一定的应用价值.     

海南东寨港真红树植物内生放线菌多样性及其抗菌活性

【目的】勘探海南东寨港真红树植物内生放线菌多样性,为发现放线菌新物种和新抗生素奠定基础。【方法】样品经表面消毒后粉碎,用10种不同培养基分离放线菌;通过PCR扩增、测定并比对16S r RNA基因序列,开展放线菌多样性分析;通过发酵、萃取等处理方法得到四类样品,包括发酵原液、乙酸乙酯提取液及水层和菌体的丙酮浸泡提取液;采用纸片扩散法对样品进行抗菌活性筛选;基于PCR的基因筛选技术探测活性菌株可能存在的NRPS、PKS I、PKS II抗生素生物合成基因。【结果】经形态特征排重,从14种真红树植物样品中共得到放线菌146株,16S r RNA基因序列比对表明它们分布于13个科18个属,其中链霉菌属为优势菌属,菌株S3Cf-2和S3Af-1的16S r RNA基因序列分别与有效发表菌株Couchioplanes caeruleus DSM44103T(X93202)和Microlunatus terrae BS6T(JF806519)的相似率最高,分别为97.45%和97.43%,可能为新物种。对其中46株放线菌发酵样品的抗菌活性检测表明,40株具有抗菌活性,总阳性率为86.96%;活性菌株中,38株菌存在至少一种所探测的生物合成基因簇,阳性率为95%,其中14株同时具有所探测的3种抗生素生物合成基因簇。【结论】海南东寨港真红树植物中存在多样性丰富的药用放线菌资源,具有从中发现放线菌新物种及新抗生素的潜力。     

Biosorption of Cd, Cu, Pb, and Zn from aqueous solutions by the fruiting bodies of jelly fungi (Tremella fuciformis and Auricularia polytricha)

In this study, dried and humid fruiting bodies of Tremella fuciformis and Auricularia polytricha were examined as cost-effective biosorbents in treatment of heavy metals (Cd²⁺, Cu²⁺, Pb²⁺, and Zn²⁺) in aqueous solution. The humid T. fuciformis showed the highest capacity to adsorb the four metals in the multi-metal solutions. The Pb²⁺ adsorption rates were 85.5%, 97.8%, 84.8%, and 91.0% by dried T. fuciformis, humid T. fuciformis, dried A. polytricha, and humid A. polytricha, respectively. The adsorption amount of Pb²⁺ by dried and humid T. fuciformis in Cd²⁺ + Pb²⁺, Cu²⁺ + Pb²⁺, Pb²⁺ + Zn²⁺, Cd²⁺ + Cu²⁺ + Pb²⁺, and Cd²⁺ + Zn²⁺ + Pb²⁺ solutions were not lower than that in Pb²⁺ solutions. The results suggested that in humid T. fuciformis, Cd²⁺, Cu²⁺, and Zn²⁺ promoted the Pb²⁺ adsorption by the biomass. In the multi-metal solutions of Cd²⁺ + Cu²⁺ + Pb²⁺ + Zn²⁺, the adsorption amount and rates of the metals by all the test biosorbents were in the order of Pb²⁺ > Cu²⁺ > Zn²⁺ > Cd²⁺. Compared with the pseudo first-order model, the pseudo second-order model described the adsorption kinetics much better, indicating a two-step biosorption process. The present study confirmed that fruiting bodies of the jelly fungi should be useful for the treatment of wastewater containing Cd²⁺, Cu²⁺, Pb²⁺, and Zn²⁺.     

青海湖水及湖滨盐化土壤放线菌的分类及耐盐性

采用3种含盐培养基(70g/L NaCl)分离青海湖水、湖泥、湖滨土壤及盐化荒地土壤中的中低温放线菌,纯化后按常规方法鉴定,并测定其耐盐性。结果表明:①湖泥、湖滨土壤及盐化荒地土壤中存在一定数量的中、低温耐盐放线菌。湖滨荒地土壤中的放线菌数量最高,其次为湖滨土壤及浅层湖水底淤泥。②鉴定出的耐盐性放线菌均为链霉菌(占供试菌总数的92.6%),且全部为白孢类群。③47.6%、19.0%和33.3%的供试放线菌的最高耐盐度分别为70、100和150g/L。     

Functional divergence of the duplicated AtKIN14a and AtKIN14b genes: critical roles in Arabidopsis meiosis and gametophyte development

Gene duplication is important for gene family evolution, allowing for functional divergence and innovation. In flowering plants, duplicated genes are widely observed, and functional redundancy of closely related duplicates has been reported, but few cases of functional divergence of close duplicates have been described. Here, we show that the Arabidopsis AtKIN14a and AtKIN14b genes encoding highly similar kinesins are two of the most closely related Arabidopsis paralogs, which were formed by a duplication event that occurred after the split of Arabidopsis and poplar. In addition, AtKIN14a and AtKIN14b exhibit varying degrees of coding sequence divergence. Further genetic studies of plants carrying atkin14a and/or atkin14b mutations indicate that, although these two genes have similar functions, there is clear evidence for functional divergence. Although both genes are important for male and female meiosis, AtKIN14a plays a more critical role in male meiosis than AtKIN14b . Moreover, either one of these two genes is necessary and sufficient for gametophyte development, indicating that they are redundant for this function. Therefore, AtKIN14a and AtKIN14b together play important roles in controlling plant reproductive development. Our results suggest that the AtKIN14a and AtKIN14b genes have retained similar functions in gametophyte development and female meiosis, but have evolved partially distinct functions in male meiosis, with AtKIN14a playing a more substantive role.     

Tyroserleutide tripeptide affects calcium homeostasis of human hepatocarcinoma BEL-7402 cells

This study aimed to observe the effects of tyroserleutide (tyrosyl-seryl-leucine, YSL) on the growth of human hepatocarcinoma BEL-7402 that was transplanted into nude mice, and explore its anti-tumor mechanism preliminarily. YSL, at doses of 80 μg-kg^-1 · d^-1, 160 μg·kg^-1 ·d^-1 and 320 μg · kg^-1 · d^-1 significantly inhibited the growth of the human hepatocarcinoma BEL-7402 tumor in nude mice, producing inhibition of 21.66%, 41.34%, and 34.78%, respectively. Ultra structure of BEL-7402 tumor in nude mice showed that YSL could induce tumor cells apoptosis and necrosis, cell organelle mitochondria and endoplasmic reticulum damage, and calcium overload. By confocal laser scanning microscopy and flow cytometry, we found that 10 μg/mL YSL rapidly induced an increase of the concentration of cytoplasmic free calcium in BEL-7402 cells in vitro, and maintained high concentrations of cytoplasmic free calcium for 1 h. Then the calcium concentration began to decrease after 2 h, and was lower than that of the control group at 4 h and 24 h (P< 0.05). YSL also decreased the mitochondrial transmembrane potential of BEL-7402 cells in vitro, but had no effect on the calcium homeostasis or mitochondrial transmembrane potential of Chang liver hepatocytes. So affecting calcium homeostasis, then inducing apoptosis and necrosis may be a mechanism by which YSL inhibits the tumor growth in animal model.     

心肺压力感受器持续卸荷对前臂血流、血管阻力及心率变异性谱的影响

低压值下体负压（ＬＢＮＰ）可仅使心肺压力感受器卸荷。采用－２ｋＰａＬＢＮＰ实验结果表明：ＬＢＮＰ既不引起动脉血压变化,也不引起心率改变,但却引起基础胸阻抗（Ｚ。）从对照的２１．８±０．４升高到２２．５±０．５Ω（Ｐ＜０．０１）,前臂血管阻力（ＦＶＲ）从１２．３±０．９升高到１９．９±１．４Ｕ（Ｐ＜０．０１）,前臂血流（ＦＢＦ）从对照时７．１±０．５降低到４．３±０．３ｍｌ·ｍｉｎ￣（－１）·１００ｍｌ￣（－１）,心率变异性谱（ＨＲＶ）未发生任何变化,即心肺压力感受器卸荷时心脏自主神经活动水平与均衡性不受影响。由于ＦＶＲ和ＦＢＦ的变化可间接反映外周血管交感传出活动水平,上述实验结果提示,心肺压力感受器对外周血管及心脏自主神经活动的调节可能存在机能分化现象。     

侧脑室注射R-α-四基组胺对哮喘豚鼠呼吸道P物质样免疫反应物的影响

目的　应用免疫组织化学方法观察侧脑室注射组胺H3 受体激动剂R α 甲基组胺对哮喘豚鼠气道内P物质样免疫反应物的影响。方法　豚鼠首先腹腔注射卵蛋白致敏 ,给药组给予豚鼠股静脉注射卵蛋白激发哮喘发作后 ,再侧脑室注射组胺H3 受体激动剂R α 甲基组胺 (5 μg)。哮喘组豚鼠在诱发哮喘发作后侧脑室注射等量生理盐水。对照组豚鼠腹腔注射等量生理盐水。经肺动脉灌注 4 %多聚甲醛 ,取气管、支气管和肺组织 ,冰冻切片 ,做P物质免疫组织化学染色。结果　气管、支气管和肺泡间隔内的P物质样免疫反应物在哮喘发作组明显多于对照组 ,在给药后 10min组明显少于哮喘组 ,给药后 2 0min组已接近正常组。结论　经侧脑室途径激动中枢组胺H3 受体后 ,哮喘豚鼠呼吸道内神经源性炎症介质P物质减少 ,提示中枢组胺H3 受体参与对哮喘气道神经源性炎症的调控。