酵母蛋白饲料培养条件的研究

对啤酒酵母进行发酵试验,结果表明:该酵母最优的培养条件是麦芽汁培养基稀释倍数2.5,培养时间20h,蔗糖添加量10Brix,尿素添加量0.02%,且酵母菌耐酸耐铜的性能好。     

酿酒酵母菌胞内海藻糖提取工艺参数的优化

建立了超声破碎酿酒酵母细胞和冷三氯乙酸化学浸提海藻糖的工艺，探索了离子色谱分析技术条件，采用响应曲面法（RSM），研究了液料比R、超声功率W、工作时间T1、超声总时间T2和浸提时间T3等5个试验关键因子对海藻糖提取的影响规律，并构建了动态控制的数学模型，得到了海藻糖提取最优工艺参数依次为：尺为7、W为698W、T1为4．9s、T2为7．3min、T3为9min。结果表明：模型极显著，具有可行性和有效性，超声功率、工作时间和超声总时间对海藻糖提取量的影响最大，为生产中的应用提供了科学依据。     

塔式罐连续培养面包酵母研究

使用塔式生化反应器连续培养面包酵母，对发酵液进行恒浊和恒化控制，并与间歇面包酵母培养法比较，结果表明，在４６Ｌ试验室发酵罐中，以稳态时平均稀释速率０．２－０．３ｈ＾－１和通气速率４．８ｍ＾３／ｈ～５．２ｍ＾３／ｈ培养面包酵母为佳，连续稳态时间平均可达４０ｈ左右。干酵母对糖得率，连续培养法比间歇法培养略有增加，活力和间歇法相当，而生产速率增加了３－４倍。     

高产单细胞蛋白酵母的诱变育种及培养条件优化

从啤酒废酵母泥中分离纯化出5株酿酒酵母,以其中单细胞蛋白(SCP)含量较高的SY-5为出发菌株,通过紫外诱变分生孢子,经过初筛和复筛,得到1株高产SCP的诱变菌株SY-5-7,比出发菌株高7.84%,达到48.67%。通过单因素试验对诱变菌株SY-5-7培养条件进行优化,结果表明,最适培养条件为培养基起始pH值5.0、培养温度26℃、接种量2%、发酵周期为70h,碳源为50g/L蔗糖、氮源为10g/L酵母膏。在此基础上进行培养,SY-5-7的生物量比初始条件提高了约35%。     

应用荧光偏振法测定酿酒酵母完整细胞膜的流动性

以酿酒酵母菌单倍体T-27(αtrp-ura-)及其耐盐突变株T-27-12(αtrp-ura-)为试验对象,考查了在高盐(25%NaCl)冲击条件下菌体生物量,并应用荧光偏振法对冲击条件下菌体细胞膜的流动性进行测定,试验数据表明:高盐冲击条件下T-27-12(αtrp-ura-)生物量明显高于T-27(αtrp-ura-),耐盐突变株T-27-12(αtrp-ura-)细胞膜的稳定性明显高于其亲株T-27(αtrp-ura-),说明T-27-12(αtrp-ura-)较其亲株有较强的抵御恶劣环境的能力,因而可以推断细胞膜的稳定性是耐盐突变株较其亲株有更强抵御高盐冲击的一个重要原因。     

酿酒酵母中磷脂合成相关基因突变对细胞自噬和液泡形态的影响

[目的]探讨酿酒酵母中磷脂合成相关基因突变对细胞自噬和液泡形态的影响.[方法]通过尼罗红染色观察酵母中磷脂合成相关基因突变后脂滴的形态;用绿色荧光蛋白(GFP)标记细胞自噬蛋白Atg8后,采用荧光显微镜和免疫印迹试验检测突变体细胞自噬的发生情况,并使用荧光染料FM4-64检测液泡形态;此外,检测相关突变体对吩嗪-1-羧酸(申嗪霉素)的敏感性.[结果]在营养有限条件下,酿酒酵母中磷脂合成相关基因突变体中脂滴的大小或数量受到不同程度的影响,但细胞自噬在常规检测条件下正常;其中磷脂酸胞苷转移酶编码基因CDS1突变导致液泡形态异常(碎片化),而其他磷脂合成相关基因突变不影响液泡形态;回补CDS1后,cds1-DAmP突变体中液泡形态恢复正常;此外,cds 1-DAmP突变体对吩嗪-1-羧酸处理更为敏感.[结论]酿酒酵母中脂滴和液泡形态异常不一定影响细胞自噬的正常进行,但可能影响其对外界环境的响应.     

酵母SOD1基因缺失突变体应答真菌细胞壁抑制剂CFW的转录组学分析

以酿酒酵母为研究材料,采用酵母全基因组表达谱芯片,分析超氧化物歧化酶SOD1基因缺失(sod1Δ)对酵母细胞应答真菌细胞壁抑制剂钙荧光白(CFW)全基因组转录表达谱的影响,为揭示植物病原真菌细胞壁调控机制以及植物抗真菌基因工程改造提供新的理论基础。结果表明:CFW(10μg/m L)处理1 h后,与野生型酵母细胞相比,sod1Δ酵母细胞中211个基因发生了显著差异表达(97个基因表达上调、114个基因表达下调)。随机选取5个差异表达基因采用定量PCR验证,结果与芯片分析结果一致。差异表达基因功能主要涉及细胞壁、细胞代谢、蛋白质合成、细胞防御以及大量功能未知蛋白。以上结果表明,SOD1基因缺失可显著改变酵母细胞应答真菌细胞壁抑制剂CFW胁迫的全基因组转录表达谱。     

啤酒酵母葡聚糖对断奶仔猪生产性能及淋巴细胞转化率的影响

为了探讨啤酒酵母葡聚糖在断奶仔猪日粮中的适宜添加剂量及其对断奶仔猪细胞免疫功能的影响,本研究进行了2个试验。试验1选用100头(28±2)d断奶的二元杂交断奶仔猪,按单因子试验设计随机分为5个处理,分别饲喂含葡聚糖0、25、50、100mg/kg和200mg/kg的日粮。结果表明:随葡聚糖添加剂量的增加,平均日增重在14￣28 d及0￣28 d呈二次曲线变化(P<0.05)。试验2选用80头(28±2)d断奶的二元杂交断奶仔猪,随机分为2个处理,分别饲喂含葡聚糖0mg/kg和50mg/kg的日粮。在试验的第14天和第28天,每重复取1头仔猪前腔静脉采血,测定外周血淋巴细胞转化率。结果显示,在断奶仔猪日粮中添加50mg/kg葡聚糖提高了仔猪在14￣28d及0￣28d的日增重(P<0.05)。而且,也提高了仔猪在0￣14 d、0￣28 d及28￣35 d的平均日采食量(P<0.05)。但是对淋巴细胞转化率没有影响。结果表明:在断奶仔猪日粮中添加50mg/kg啤酒酵母葡聚糖,可以提高断奶仔猪的生产性能,而且没有性别差异。     

不同酵母发酵的可可果酒香气成分分析

以可可鲜豆为原料,选取酿酒曲増香高产型、葡萄酒/果酒专用型、32762、1425、1557、葡萄酒/果酒高活性型、1793共7种酿酒酵母进行发酵,利用固相微萃取和气质联用技术分别对7种可可果酒不同发酵阶段的香气成分进行比较。结果表明:发酵2周时,酒液中检出的香气成分分别为41、47、37、39、36、50、35种,其中葡萄酒/果酒高活性型酿制的果酒中香气成分种类较多;可可果酒中香气成分以醇类和酯类为主,主要的酯类成分为辛酸乙酯、癸酸乙酯、苯乙酸乙酯、乙酸苯乙酯、9-十六碳烯酸乙酯,醇类中乙醇和苯乙醇存在较多,其中苯乙醇为呈香成分,这些特征香气成分赋予了可可果酒独特的风味。通过仪器测定和嗅闻感官综合判断,葡萄酒/果酒专用型、葡萄酒/果酒高活性型发酵的可可果酒香气和口感较好,适宜大众口味。     

A Novel Particle Contact Assay with the Yeast Saccharomyces cerevisiae (8 pp)

Goal, Scope and Background   Numerous xenobiotics released into surface waters are transferred to suspended particulate matter and finally attached to sediments. Aquatic organisms may be exposed to them by direct particle feeding, by physical contact with contaminated surfaces as an exposure route, and by the uptake of dissolved contaminants after equilibration via the free water phase. In order to assess potential sediment toxicity, each of these exposure routes has to be addressed. This paper presents a newly developed particle contact assay that uses the fermentation performance of a specific Saccharomyces cerevisiae strain for the assessment of toxic effects in sediments. The test procedure is based on the characteristic feature of growing yeast cells to attach to sediment particles, which are also relevant for the accumulation of contaminants. The physical contact with lipophilic contaminants mirrors an exposition pathway for the direct uptake into the cells. In order to quantitatively characterize the toxic effects of particle attached pollutants on the fermentation performance, unpolluted native reference sediment was spiked with representatives for widely distributed anthropogenic contaminants. Methods   Saccharomyces cerevisiae was established as sensitive eukaryotic microorganism for the ecotoxicological assessment of particle attached anthropogenic contaminants in freshwater sediments. For this purpose, yeast cells were cultivated in sediment samples and the resulting fermentation performance was continuously measured. Sediments artifically spiked with HCB, PCB, g-HCH, DDT, and benzo(a)pyrene and solutions of each contaminant were comparatively investigated by means of their adverse effects on yeast fermentation performance. Additionally, four native river sediments characterized by increasing levels of pollution were assessed by the yeast particle contact assay, and simultaneously by standard aquatic tests with algae, daphniae, and luminescent bacteria using pore water and elutriates. Results of the bioassays were related to specific sediment contamination with respect to metals and organic priority pollutants. Results and Discussion   In sediments spiked with PCB and benzo(a)pyrene fermentation, performance was affected extensively below concentrations inhibiting fermentation in contaminant solutions. This suggests a high efficiency of the exposure route by physical contact. The fermentation performance was only slightly affected by single lipophilic pollutants, whereas mixtures of individually spiked sediments caused critically reduced fermentation performance suggesting additive synergistic effects. Native river sediments modestly to critically polluted by hazardous organic compounds lead to a slightly to dangerously reduced fermentation performance in the yeast contact assay. These inhibitory effects were much less pronounced in the standard bioassays conducted with algae, daphniae and luminescent bacteria, applying pore waters and elutriates as sample matrices. Using pore water, inhibition was measured only in the most polluted sediment, elutriates lead to a slight inhibition of the algal growth in the undiluted sample only. These results indicate an improved sensitivity of the yeast particle contact assay compared to the standard assays, due to uptake and physical cell contact as additional routes of exposure. Conclusion   The yeast particle contact assay is a valuable tool for the assessment of ecotoxicological potential in freshwater sediments. Since the assay addresses physical contact as an exposure route, it indicates bioavailability of lipophilic compounds in sediments. Outlook   The sensitive indication of bioavailable contaminants associated to sediment particles by the newly developed yeast particle contact assay recommends it as a complementary microbial bioassay in a test battery for assessing major pathways of contaminants in whole sediments.