Insights into the subunit interactions of the chloroplast ATP synthase

Subunit interactions of the chloroplast F0F1- ATP synthase were studied using the yeast two-hybrid system. The coding sequences of all the nine subunits of spinach chloroplast ATP synthase were cloned in two-hybrid vectors. The vectors were transformed into the yeast strains HF7c and SFY526 by various pairwise combinations, and the protein interactions were analyzed by measuring the yeast growth on minimal SD medium without serine, lucine and histidine. Interactions of γ Subunit with wild type or two truncated mutants of γ sununit, △εN21 and △εC45, which lose their abilities to inhibit the ATP hydrolysis, were also detected by in vitro and in vivo binding assay. The present results are largely accordant to the common structure model of F0F1-ATP synthase. Different from that in the E. Coli F0F1-ATP synthase, the δ subunit of chloroplast ATP syn- thase could interact with β,γ,ε and all the CF0 subunits in the two-hybrid system. These results suggested that though the chloroplast ATP synthase shares the similar structure and composition of subunits with the enzyme from E. Coli, it may be different in the subunit interactions and con- formational change during catalysis between these two sources of ATP synthase. Based on the present results and our knowledge of structure model of E. Coli ATP synthase, a deduced structure model of chloroplast ATP synthase was proposed.     

Vacuolar H+-ATPase: From mammals to yeast and back

Vacuolar H⁺-adenosine triphosphatase (V-ATPase) is composed of distinct catalytic (V₁) and membrane (V₀) sectors containing several subunits. The biochemistry of the enzyme was mainly studied in organelles from mammalian cells such as chromaffin granules and clathrin-coated vesicles. Subsequently, mammalian cDNAs and yeast genes encoding subunits of V-ATPase were cloned and sequenced. The sequence information revealed the relation between V- and F-ATPases that evolved from a common ancestor. The isolation of yeast genes encoding subunits of V-ATPase opened an avenue for molecular biology studies of the enzyme. Because V-ATPase is present in every known eukaryotic cell and provides energy for vital transport systems, it was anticipated that disruption of genes encoding V-ATPase subunits would be lethal. Fortunately, yeast cells can survive the absence of V-ATPase by drinking the acidic medium. So far only yeast cells have been shown to be viable without an active V-ATPase. In contrast to yeast, mammalian cells may have more than one gene encoding each of the subunits of the enzyme. Some of these genes encode tissue- and/or organelle-specific subunits. Expression of these specific cDNAs in yeast cells may reveal their unique functions in mammalian cells. Following the route from mammals to yeast and back may prove useful in the study of many other complicated processes.     

酵母菌处理味精废水的研究

对酵母菌处理味精废水的工艺条件进行了研究．结果表明，其最佳工艺条件为：pH4．0、温度32℃、发酵时间22h、接种量15％．该技术作为前处理工艺可使废水化学耗氧量(COD)的去除率达60％以上，为后处理的达标排放提供了基础，并可回收一定的酵母蛋白，具有一定的经济效益。     

铬酵母制备及在酸奶中的添加

利用生物合成法制备了有机铬含量较高的新型添加剂——铬酵母，并将其添加到酸奶中．用分光光度法对铬酵母及添加了铬酵母的酸奶进行了总铬和有机铬含量的测定，考察了样品处理方法，并从酸奶的感官鉴评、酸度及有机铬含量方面考察铬酵母的添加对酸奶品质的影响．结果表明，铬酵母的添加对酸奶品质无任何不良影响，而有机化程度在发酵前后也无明显的变化．     

糖蜜发酵培养高铁营养酵母

采用生长周期短，易于工业化生产的酵母为菌株，以糖蜜为主要培养基，通过现代发酵工程技术，在3m^3发酵罐上进行中间试验，在12m^3发酵罐上试生产高铁营养酵母．其发酵产品高铁营养酵母铁的含量为1061mg／kg，高铁营养酵母的产率为2．54％，蛋白质的含量(质量分数)为51.90％.     

葡萄糖-6-磷酸脱氢酶的分离纯化

报道了葡萄糖-6-磷酸脱氢酶的分离纯化. 在0.1 mol/L的碳酸氢钠溶液中,以溶胀法从酵母中提取胞内酶.粗抽提液经过60 %～75 % 饱和度的硫酸铵盐析, Sephadex G-100凝胶柱层析.葡萄糖-6-磷酸脱氢酶的收得率为12.4%,纯化倍数达51.2.     

酵母细胞流加培养过程的建模与预估控制

建立了包括菌体浓度 X、底物浓度 S、培养液体积 V为状态变量的酵母流加发酵过程的非结构动力学模型 ;基于该模型 ,提出了一种克服被估变量的累积误差的一步预估方法 ,用以估计难以可在线测量的菌体浓度、底物浓度等生化状态变量 ,并根据预估结果 ,进行流加操作。该方法应用于酵母流加培养 ,酵母对糖得率由 35 .1%提高到 40 .2 %。     

酵母海藻糖提取及精制工艺的研究

对酵母海藻糖的提取与纯化工艺进行了研究。在温度80℃、酵母质量浓度70g/L条件下用50%乙醇溶液提取60min,海藻糖提取率可达98.81%;采用大孔阴阳离子交换树脂混合柱纯化提取液,适宜条件为,流速(3～6)mL/min,温度40℃,经浓缩、结晶,海藻糖纯度可达98%以上。     

不同浓度的硫离子处理酵母菌的生物学效应

将不同浓度的Na2S加入培养基中处理酵母菌,结果发现,随着处理培养基中Na2S浓度的升高,酵母菌的菌落数、细胞的直径、细胞内多种内含物（核酸、蛋白质、可溶性糖等）的含量都有所降低.     

新型抗菌肽基因设计、合成及在酵母中的表达(Ⅱ)——抗菌肽 AD 基因在酵母中的表达

将抗菌肽ＡＤ基因定向克隆到大肠杆菌－酵母穿梭质粒ｐＣＬＷＡ２的ＢａｍＨＩ和ＳａｌＩ位点上,构建成含抗菌肽ＡＤ基因的重组质粒ｐＣＡＤ,转化酵母宿主菌ＡＢ１０３,转化子在缺乏亮氨酸的ＳＤ选择培养中３０℃培养４８ｈ,培养液经简单纯化后,活性蛋白ＰＡＧＥ和琼脂孔穴扩散法测定抑菌活性表明,抗菌肽ＡＤ基因在酵母中获得表达．