橙汁果酒绮丽酵母的筛选及耐受性

为获得适合橙汁果酒发酵的菌种,从橙汁发酵醪液中纯化出酵母菌,采用杜氏管法、CO2失重法及感官评定法进行筛选。通过形态观察和生理生化试验对菌株进行鉴定,并对菌株进行耐受性研究。结果表明：从发酵醪液中分离出4株酵母菌,经过初筛及复筛得到一株发酵能力强、还原糖利用率高、果酒风味柔和等优良特征的酵母菌,编号为 FJ-20,鉴定为酵母属的绮丽酵母。该菌株对酸的耐受能力为 pH2．5,对酒精的耐受能力为10％vol,对氯化钠耐受能力为11％,对蔗糖的耐受能力为35％。该菌株发酵能力强、还原糖利用率高、果酒风味柔和,发酵产香浓郁,适合作为橙汁低纯果酒的酿造酵母。     

Construction of Recombinant Expression Plasmid pYELmleA of mleA Gene and Expression in Saccharomyces cerevisiae

苹果酸-乳酸酶是进行MLF的功能酶。笔者进行酒酒球菌SD-2a的苹果酸-乳酸酶基因重组表达质粒的构建。利用来自重组质粒pLmleA的，mleA基因，以PGK1强启动子和ADH1终止子为调控元件。以大肠杆菌-酵母菌穿梭质粒YEp352为载体，构建了重组表达质粒pYELmleA，并转化酿酒酵母(Saccharomyces cerevisiae)YS58。酵母转化子用含有亮氨酸、组氨酸和色氨酸的YNB平板筛选鉴定。SDS—PAGE检测表明获得的转化子表达了约60KD的目标蛋白。斑点杂交检测表明目的基因mleA转化到受体菌中。获得的转化子在添加了L-苹果酸的培养基中培养4d；取培养液上清用HPLC检测L-苹果酸及L-乳酸含量，采用t检验进行差异显著性分析，结果表明mleA基因进行了功能性的表达，将L-苹果酸转化成L-乳酸，L-苹果酸和L-乳酸含量分别与对照差异极显著和显著，L-乳酸的生成量为1002—1106mg／L。苹果酸的相对降低率为19．16—22．34％。     

Effects of Saccharomyces cerevisiae on in vivo organic matter digestibility and milk yield in buffalo cows

The influence of a dietary supplementation with Saccharomyces cerevisiae (SC) during the first and the second phase of lactation on dry matter (DM) intake, organic matter digestibility, milk yield and quality and haematological profile was evaluated in buffalo cows. Lactating buffaloes (n = 190), 118.7 days in milk (DIM), were randomly divided into Group C (control, n = 95) and Group T (fed diet supplemented with 98 billion CFU of S. cerevisiae, n = 95). Eight buffaloes for each group (Groups T1 and C1), 85.4 DIM, were used to study the in vivo digestibility and the haematological profile. No differences were found for DM intake (16.5 kg·day^− 1) and haematological profile. The SC supplementation increased milk yield (7.9 ± 0.2 vs. 7.4 ± 0.2; P < 0.01) but did not affect milk fat and protein. SC supplementation increased OM digestibility, mainly, in the first phase of lactation (< 135 days), thus allowing a higher energy availability for milk yield and reduced fat mobilization.     

几种益生菌发酵谷物饮料制作工艺研究

益生菌发酵谷物饮料既保存了谷物原有的营养价值,又具有益生茵发酵制品的有益作用,风味良好、口感独特,对人体有显著的营养保健作用.用1.5%的量接种从发酵谷物中筛选获得的酿酒酵母、嗜酸乳杆菌、保加利亚乳杆菌分别发酵糯米制作米酒、发酵玉米汁制作酸性饮料、多肽饮料,其发酵条件分别为:米酒发酵温度30℃,发酵时间72h,酸性饮料...     

南阳酵母原生质体制备与再生条件研究

研究南阳酵母原生质体形成与再生的最佳条件。获取原生质体采用酶法,再生采用双层高渗平板法,因素重要性分析采用正交试验。结果表明:菌龄10h,以2.0%蜗牛酶加0.5%纤维素酶的混合酶液进行酶解破壁,酶解温度30℃,酶解时间2.5h,以0.6mol.L-1蔗糖做再生培养基的渗透压稳定剂,原生质体形成率91.40%,再生率10.56%。不进行预处理,在试验条件下,对南阳酵母原生质体形成与再生影响因素依次为:菌龄>酶解时间>酶浓度>渗稳剂。     

The Nutritive Value For Growing Pigs of Single Cell Protein (Saccharomyces Cerevisiae) Produced from Sulphite Spent Liquor

The effects on growing pigs of substituting 4.5 and 9 % of soy bean meal with a corresponding amount of single cell protein produced from sulphite spent liquor in a diet based on cereals and fish meal have been studied. The concentration of lignosulphonic acids in the single cell protein product was found to be 0.6 % ± 0.2 % (m. ± s). No differences in the weight gain, feed conversion ratio or fat thickness of the pigs, as compared with the controls, were observed when fed single cell protein-containing diets from about 29 to 80 kg in the course of 11 weeks. Nor were any effects found on addition of 0.15 % of methionine to the diets.     

六个甘蓝型油菜油酸脱氢酶（FAD2）假基因的克隆和分析

以甘蓝型油菜湘油15为材料,采用PCR方法克隆并分析了56个FAD2基因克隆和47个FAD2基因cDNA克隆,从中发现6个新的FAD2基因拷贝。它们与公布的甘蓝型油菜FAD2基因(AY577313)具有87%以上的同源性,没有内含子,在开放阅读框中存在1~12个终止密码子,其中有2个拷贝具有转录功能。将这6个FAD2基因拷贝在酿酒酵母中进行体内表达实验,通过气相色谱检测脂肪酸组成证明其不具备油酸脱氢酶功能,与对照相比,也没有改变酵母体内脂肪酸组成。由此推测这6个FAD2基因拷贝为假基因。     

Re-examination of inhibitor resistance conferred by Qo-site mutations in cytochrome b using yeast as a model system

Cytochrome b from yeast (Saccharomyces cerevisiae Meyer ex Hansen) provides a convenient model system for the study of Qo-site inhibitor (QoI) resistance mutations from a variety of organisms. QoI resistance mutations from fungal plant pathogens (G143A and F129L), malaria agent Plasmodium sp (Y279C/S), and Pneumocystis carinii (L275F), an opportunistic pathogenic fungus of man, were introduced into yeast cytochrome b and their effect on the binding of a variety of natural (myxothiazol and stigmatellin) and synthetic (atovaquone, azoxystrobin and pyraclostrobin) inhibitors to the bc1 complex monitored. L275S (from a myxothiazol-resistant yeast) was also re-examined. Stigmatellin binding was relatively unaffected by the introduction of these mutations. Significant increases in resistance were observed for the strobilurin-class inhibitors myxothiazol, azoxystrobin and pyraclostrobin, with the largest increase in resistance conferred by G143A. In contrast, atovaquone binding was most effected by Y279C/S and L275S. Notably, F129L, G143A and L275S had a minor effect on bc1 activity, and so are unlikely to confer significant fitness penalties in vivo. These data are discussed in the light of the atomic structures for myxothiazol- and azoxystrobin-inhibited bovine bc1 which have recently become available. We propose that QoI resistance due to G143A arises from steric hindrance between the inhibitor and cytochrome b, whereas the mechanism of resistance for the other mutations is due to an increase in binding energy between the protein and inhibitor molecule. Site-directed mutagenesis was also used to model selected regions of the mammalian Qo site in yeast cytochrome b in order to further understand the differential efficacy of these QoI in the mammalian and pathogen bc1 complexes.     

纳他霉素纳米乳的制备及其体外抑菌效果研究

【目的】制备纳他霉素纳米乳(NATA-NE),测定其对白色念珠菌（Candida albicans）、青霉菌（Penicillium）和酿酒酵母菌（Saccharomyces cerevisiae）的最小抑菌浓度（MIC）和最小杀菌浓度（MFC）,以指导临床合理用药。【方法】综合纳米乳和纳他霉素的优势,将纳他霉素制成水包油型NATA-NE,以酮康唑和特比萘芬作为阳性药物对照,根据美国临床实验室标准化委员会（NCCLS）制定的真菌体外药敏试验方法M27A和M-38P,测定药物对3种受试菌株的MIC,并研究了NATA-NE的杀菌效果。【结果】NATA-NE对青霉菌、白色念珠菌和酿酒酵母菌的MIC值为1,0.5,0.5 μg/mL;NATA原料药、酮康唑和特比萘芬对上述3种菌的MIC值分别为2,1,1 μg/mL;4,8,8 μg/mL和1,8,16 μg/mL,NATA-NE的MIC值均小于或等于其他3种药物。3种菌在2 μg/mL NATANE的作用下1.5 h后,均未见生长;NATA原料药则需要4 μg/mL、2 h才能达到同样效果,而酮康唑和特比萘芬分别需要16 μg/mL、4 h和32 μg/mL、4 h才能达到同样效果。【结论】临床使用NATA-NE治疗动物真菌病时,可适当降低给药剂量和用药次数,以降低成本和对动物不可预期的副作用。     

酿酒酵母菌及其灭活菌对绵羊瘤胃外植体β-防御素-1(SBD-1)表达的影响

旨在探索益生性酿酒酵母菌及其灭活菌对绵羊瘤胃外植体内β-防御素-1(sheep beta-defensin-1,SBD-1)表达的影响。本研究建立了绵羊瘤胃外植体培养方法,将不同浓度(10~4、10~5、10~6、10~7、10~8、10~9 CFU·mL^-1)的酿酒酵母菌及其灭活菌与外植体共培养24 h后,通过qPCR和ELISA方法检测瘤胃外植体内SBD-1 mRNA和蛋白的表达变化,以分别确定活菌及其灭活菌诱导SBD-1表达最高的菌液浓度;然后用该浓度的活菌及其灭活菌对瘤胃外植体进行不同时间(2、4、8、12、16、20、24 h)的刺激,同样用qPCR和ELISA方法检测瘤胃外植体内SBD-1 mRNA及蛋白的表达变化,从而筛选出活菌及其灭活菌诱导SBD-1表达的最佳时间。结果表明,益生性酿酒酵母菌及其灭活菌均能显著促进绵羊瘤胃外植体SBD-1的表达(P<0.05)。当酿酒酵母菌浓度为10~7 CFU·mL^-1、灭活菌浓度为10~8 CFU·mL^-1分别诱导绵羊瘤胃外植体16 h时,SBD-1表达量分别达到最大,且二者与对照相比均呈极显著差异(P<0.01)。在最佳诱导条件下对比二者的诱导效果,酿酒酵母菌活菌高于其灭活菌。因此,益生性酿酒酵母菌及其灭活菌均能促进绵羊瘤胃外植体内SBD-1的表达,且活菌浓度为10~7 CFU·mL^-1、灭活菌浓度为10~8 CFU·mL^-1分别诱导16 h时,绵羊瘤胃外植体内的SBD-1的表达量分别达到最大,并且益生性酿酒酵母菌的诱导效果高于其灭活菌。