糖化酵母在酒精工业中应用的研究 (Ⅱ)——原生质体融合构建能分解淀粉的高温酒精酵母

利用ＰＥＧ诱导原生质体融合技术，进行糖化酵母单倍体２６００７（α，ＳＴＡ，ｉｎｈ，ａｄｅ）和去除了ＩＮＨ１基因的耐高温酒精酵母单倍体菌株Ｚ６－１０（α，ＳＴＡ，ｉｎｈ，ａｒｇ）的同型原生质体融合实验，在再生基本培养基上挑选融合子，获得一株既具有较强糊精利用能力，又耐高温和个产酒精的融合株Ｆ４１。经与亲本酒精酵母二倍体菌株２１００２５玉米发酵性比较，使用Ｆ４１菌发酵可减少４０％的糖化酶用量。     

耐高渗酵母发酵生产甘油过程中杂菌污染的探讨

从菌种培养、生产发酵、流加补料等方面初步探讨了发酵生产甘油过程中杂菌污染的途径。     

麦秸粉酵母饲料的实验室固体发酵工艺研究

研究确定了在实验室中酵母发酵麦秸粉的主要培养条件，并对发酵前后物料的粗蛋白，真蛋白，粗纤维等指标进行了测定。     

1,6—二磷酸果糖生产条件的优化控制

在用啤酒酵母发酵生产１，６－二磷酸果糖的过程中，通过优化控制过程中的ｐＨ温度，糖及磷酸盐含量，搅拌速度等条件可使ＦＤＰ的生成速率以及磷的转化率达到适应工业化生产的水平。     

抗HBsAg单链Fab抗体基因酵母表达载体的构建

采用重叠PCR技术 ,以抗乙肝表面抗原 (HBsAg)IgG铰链区基因为Linker将Fab抗体基因的重链和轻链连接起来 ,构成单链Fab基因。通过测序鉴定 ,克隆的单链Fab基因与理论上的完全一致 ,并成功构建含完整单链Fab基因的毕赤酵母 (P pastoris)表达载体。     

利用啤酒废酵母进行营养型酵母调味品的研究

研究利用啤酒废酵母为原料生产营养型酵母调味品的方法和条件。原料经洗涤、脱苦、自溶、真空浓缩等工序得到含多种营养成分的 ,具有增鲜、增香赋予食品肉香味等功能的、符合国家理化、卫生指标的调味品。该工艺的最佳条件为 :用 0 .5 % (w/w)NaHCO3溶液脱苦0 .5h ,自溶加水量为 2倍 ,自溶剂为 1.0 % (w/w)的食盐并在 (45± 1)℃条件下自溶 4 8h .     

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.     

Formation of 4,4′-methylene-bis(2-chloroaniline)-DNA adducts in yeast expressing recombinant cytochrome P450s

N-Oxidation of 4,4-methylene-bis(2-chloroaniline) (MBOCA) may lead to formation of DNA adducts. To determine if cytochrome P450s are involved in the formation of MBOCA derived-DNA adducts, yeast strains expressing rodent P450s were exposed to MBOCA, and³²P-postlabelling of nucleotides from yeast genomic DNA was done. Chromatographic analysis on PEI cellulose showed that, upon exposure to MBOCA for 1 h, nine DNA adducts were formed in yeast expressing phenobarbital-inducible rabbit P450 2B5. With a 4-h-exposure, all adducts increased in parallel. In cell-free experiments, the incubation of MBOCA with phenobarbital-induced rat microsomal fraction followed by incubation with thymus DNA, led to the formation of more than ten DNA adducts. When yeast expressing 3-methylcholanthrene-inducible rat P450 1A1 was exposed to MBOCA, one major and two minor adducts were formed. No adducts were detected in control yeast. These results show that recombinant rabbit P450 2B5 exhibits a potential activation of MBOCA and that rat P450 1A1 has some effect. The use of yeast expressing recombinant P450s and the technique of³²P-postlabelling facilitates a simple search for chemicals with carcinogenic potential.     

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。