Cofactor and CO2 donor regulation involved in reductive routes for polymalic acid production by Aureobasidium pullulans CCTCC M2012223

Polymalic acid (PMA) is a water-soluble polyester with many attractive properties for biomedical application. Its monomer l-malic acid is widely used in the food industry and also a potential C4 platform chemical. Cofactor and CO₂ donor involved in the reductive routes were investigated for PMA production by Aureobasidium pullulans. Biotin as the key cofactor of pyruvate carboxylase was favor for the PMA biosynthesis. Na₂CO₃ as CO₂ donor can obviously improved PMA titer when compared with no CO₂ supplier NaOH, and also exhibit more advantages than the other donor CaCO₃ because of its water-soluble characteristic. A combinational process with addition of biotin 70 mg/L and Na₂CO₃ as the CO₂ donor was scaled-up in 50 L fermentor, achieving the high product 34.3 g/L of PMA and productivity of 0.41 g/L h. This process provides an efficient and economical way for PMA and malic acid production, and is promising for industrial application.     

植物微核技术在监测水质污染中的应用

应用韭菜微核技术,检测哈尔滨市量具刃具厂含铬废水的遗传毒性,试验表明：含铬废水可诱导韭菜和蚕豆根尖细胞产生微核,进行回归分析,其蚕豆对Ｃｒ＾＋６诱变剂回归分析为ｙ＝２４．９４＋１２７．７７ｘ（ｒ＝０．９９６,Ｐ〈０．０１）,韭菜对Ｃｒ＾＋６诱变剂回归广泛为：ｙ＝２１．７４＋１００．２３ｘ（ｒ＝０．９００,Ｐ〈０．０１）,ｒ＝１１．１７＋３．３８ｘ（ｒ＝０．７８１,Ｐ〈０．０５）,韭菜根尖微核技术具     

洞察景观环境影响蜜蜂之新视角: 肠道微生物

作为生态服务提供者的传粉蜜蜂与景观生态息息相关, 而以农田为主的景观组成显著降低了传粉蜜蜂的多样性。目前调查研究显示, 农田的扩张与蜜蜂多样性下降相关, 且农药残留对蜜蜂损害严重。景观中的开花植物决定了蜜蜂的食物(营养)组成, 其中花粉蛋白含量与蜜蜂的生长发育紧密相关。尽管研究已证实景观环境会显著影响蜜蜂蜂群的发展和个体的生长繁殖能力, 但未来还需要加强景观组成变化直接作用于蜜蜂的机制研究。另一方面, 大量研究表明蜜蜂肠道共生菌是影响宿主健康的重要因素: 可促进宿主吸收营养和抵抗病原菌。作为传粉者, 蜜蜂接触到的主要外部环境——花粉和花蜜都含有特殊的微生物, 很多研究暗示花源微生物是蜜蜂肠道菌来源之一。研究表明景观环境相关的食物(营养)、农药残留以及环境微生物都会显著影响肠道微生物。现有少量的研究证明不同景观的蜜蜂肠道微生物有差异, 景观环境可能通过作用于蜜蜂肠道微生物进而影响蜜蜂健康。然而不同景观环境中的微生物, 尤其是花源微生物和蜜蜂肠道菌之间的关联有待证明。景观对蜜蜂肠道微生物的影响值得研究, 希望可以从肠道菌的视角鉴别对蜜蜂友好的景观环境, 进而指导土地合理利用和蜜蜂保护。     

用一种改进的电泳方法测定抗冻蛋白的分子量

针对抗冻蛋白分子量较小的特点,该文采用了一种改进的Ｔｒｉｃｉｎｅ－ＳＤＳ－ＰＡＧＥ法测定其分子量。采用三层胶系统并在电极缓冲液中以Ｔｒｉｃｉｎｅ代替Ｇｌｙｃｉｎｅ。     

用染料修饰吐温80液—固萃取体系从猪心肌匀浆液中分离纯化心肌黄酶

用三嗪类染料 Ｃｉｂａｃｒｏｎ Ｂｌｕｅ Ｆ３Ｇ－Ａ修饰的吐温８０,与吐温８０、硫酸被构成液－固萃取体系,从猪心肌匀浆液中分离纯化心肌黄酶。研究了吐温８０染料修饰物在吐温８０相中所占的比例、分相盐浓度、溶液的酸度、匀浆液的加入量等对匀浆液中酶及杂蛋白在两相中分配的影响。在室温条件下,酶选择性地进入吐温８０固相,杂蛋白主要留在盐水相。匀浆液中心肌黄酶的酶活力平均收得率为８１．４％,一步纯化倍数为６．６。降低盐浓度,提高盐水相酸度,能使酶从吐温８０固相反萃到盐水相。     

Impact of single-chain Fv antibody fragment affinity on nanoparticle targeting of epidermal growth factor receptor-expressing tumor cells

To determine the importance of single-chain Fv (scFv) affinity on binding, uptake, and cytotoxicity of tumor-targeting nanoparticles, the affinity of the epidermal growth factor receptor (EGFR) scFv antibody C10 was increased using molecular evolution and yeast display. A library containing scFv mutants was created by error-prone PCR, displayed on the surface of yeast, and higher affinity clones selected by fluorescence activated cell sorting. Ten mutant scFv were identified that had a 3-18-fold improvement in affinity (KD=15-88 nM) for EGFR-expressing A431 tumor cells compared to C10 scFv (KD=264 nM). By combining mutations, higher affinity scFv were generated with KD ranging from 0.9 nM to 10 nM. The highest affinity scFv had a 280-fold higher affinity compared to that of the parental C10 scFv. Immunoliposome nanoparticles (ILs) were prepared using EGFR scFv with a 280-fold range of affinities, and their binding and uptake into EGFR-expressing tumor cells was quantified. At scFv densities greater than 148 scFv/IL, there was no effect of scFv affinity on IL binding and uptake into tumor cells, or on cytotoxicity. At lower scFv densities, there was less uptake and binding for ILs constructed from the very low affinity C10 scFv. The results show the importance of antibody fragment density on nanoparticle uptake, and suggest that engineering ultrahigh affinity scFv may be unnecessary for optimal nanoparticle targeting.     

罗布麻对不同浓度盐胁迫的生理响应

利用网室盆栽实验,研究不同浓度的NaCl（100-400mmol·L^-1）胁迫对罗布麻（Apocynum venetum）生长及生理特性的影响。结果表明,100mmol·L^-1NaCl处理显著降低了罗布麻植株的鲜重,但对其干重影响不大;随着盐浓度继续增加,罗布麻鲜重和干重显著下降。在盐胁迫下,罗布麻叶片内的丙二醛含量、电解质渗漏率、根部和地上部Na^＋的含量明显增加,K^＋的含量随着盐离子浓度的增加而降低。盐胁迫显著降低了地上部Ca^2＋的含量,而对根部Ca^2＋的含量没有影响。植株K^＋/Na^＋和Ca^2＋/Na^＋比值随着盐胁迫强度的增加而降低。盐胁迫显著促进了罗布麻根部对K^＋和Ca^2＋的选择性吸收及对K^＋的选择性运输。当NaCl浓度小于或等于200mmol·L^-1时,随着盐离子浓度的增加,罗布麻叶片内的脯氨酸和可溶性糖积累显著增加,而当NaCl浓度大于200mmol·L^-1时,这2种有机溶质含量显著下降。总体上,罗布麻通过积累无机离子、合成有机溶质及维持较高的K^＋、Ca^2＋选择性吸收和运输来适应一定浓度（≤200mmol·L^-1NaCl）的盐胁迫。     

短序十大功劳基因组总DNA提取方法研究

以短序大功劳嫩叶为材料,采用CTAB法、CTAB改良法1、CTAB改良法2、SDS法和试剂盒法五种方法提取短序十大功劳基因组总DNA,用分光光度计和琼脂糖凝胶电泳方法检测所得总DNA的纯度和得率,用ISSR-PCR扩增的方法检测所得总DNA的质量。结果表明,五种方法均能从短序大功劳叶片中提取到基因组DNA,但不同方法提取得的基因组DNA的纯度、浓度和得率存在明显的差异。CTAB改良法2和试剂盒法提取的DNA纯度高,可直接用于下游分子生物学实验,CTAB法、CTAB改良法1和SDS法提取的总DNA质量较差,不利于下游的分子生物学实验;五种方法提取的总DNA的得率在10.836~451.709μg/g之间,呈CTAB法>SDS法>CTAB改良法1>CTAB改良法2>试剂盒法的现象。此实验获得的结果可以为短序十大功劳分子生物学研究提供基础。