发酵性丝孢酵母HWZ004利用水稻秸秆水解液发酵产油脂

为高效利用水稻秸秆中的纤维素和半纤维素产油脂,采用稀酸预处理和酶水解两步法对水稻秸秆进行水解,然后以水解液为碳源,培养发酵性丝孢酵母Trichosporon fermentans HWZ004产微生物油脂。结果表明,经简单overliming法脱毒后水稻秸秆水解液中乙酸、糠醛和5-羟甲基糠醛的浓度分别为0.4 g/L、0.1 g/L和0.05 g/L。只需添加少量氮源和微量CuSO4?5H2O,该水解液即可满足T. fermentans HWZ004发酵产油脂的要求。发酵最适接种量、初始pH和温度分别是5.0％、7.0和25 ℃。T. fermentans HWZ004在优化条件下培养7 d的生物量、油脂含量和油脂产量分别是26.4 g/L,52.2％和13.8 g/L;油脂得率系数为17.0,大大高于驯化前菌株T. fermentans CICC 1368在脱毒水稻秸秆半纤维素水解液中的对应值 (11.9)。所产油脂的脂肪酸组成与植物油相似,不饱和脂肪酸含量达70％以上,宜作为生物柴油的生产原料。     

苜蓿尺蠖核型多角体病毒囊膜蛋白

苜蓿尺蠖核型多角体病毒（ Ａｕｔｏｇｒａｐｈａ ｃａｌｉｆｏｒｎｉｃａ ｎｕｃｌｅａｒ ｐｏｌｙｈｅｄｒｏｓｉｓ ｖｉｒｕｓ,ＡｃＭＮＰＶ）在细胞质中合成其囊膜蛋白,但在细胞核内组装并包埋病毒粒子,这些蛋白的核定向转运机制是人们甚感兴趣的课题。以ＡｃＭＮＰＶ多角体衍生型病毒ＯＤＶ（ｏｃｃｌｕｓｉｏｎ－ｄｅｒｉｖｅｄ ｖｉｒｕｓ,ＯＤＶ）的一种囊膜蛋白ＯＤＶ－Ｅ１８为对象,通过Ｅ１８与一个标记短肽Ｆ     

Modeling of yeast metabolism and process dynamics in batch fermentation

Much is known about yeast metabolism and the kinetics of industrial batch fermentation processes. In this study, however, we provide the first tool to evaluate the dynamic interaction that exists between them. A stoichiometric model, using wine fermentation as a case study, was constructed to simulate batch cultures of Saccharomyces cerevisiae. Five differential equations describe the evolution of the main metabolites and biomass in the fermentation tank, while a set of underdetermined linear algebraic equations models the pseudo-steady-state microbial metabolism. Specific links between process variables and the reaction rates of metabolic pathways represent microorganism adaptation to environmental changes in the culture. Adaptation requirements to changes in the environment, optimal growth, and homeostasis were set as the physiological objectives. A linear programming routine was used to define optimal metabolic mass flux distribution at each instant throughout the process. The kinetics of the process arise from the dynamic interaction between the environment and metabolic flux distribution. The model assessed the effect of nitrogen starvation and ethanol toxicity in wine fermentation and it was able to simulate fermentation profiles qualitatively, while experimental fermentation yields were reproduced successfully as well.     

固定化生长酵母连续生产酒精的研究中间试验报告

造粒器与柱式生物反应器成—封闭系统,采用正交试验确定的最适固定化条件,以海藻酸钙凝胶法进行细胞固定化,连续造粒,在反应器中完成固定化。固定化酵母在反应器中通气培养20小时左右,凝胶球细胞数达2×10~9/me,其增长速度比传统工艺自然细胞快10倍。固定化生长细胞用于生物反应器连续发酵甜菜糖蜜酒精,酒精生产能力为22.1—23.67g/L凝胶球/小时,为传统工艺酒精生产能力的10倍。停留时间为3小时。反应器系统由两个0.7KL柱式反应器和1个0.8KL成熟醪接收器组成。发酵周期由传统工艺的20小时左右,缩短为4小时,酒精含量为8.5—9.0％(V/V),对1.2号反应器的动态变化及其在发酵中的作用进行了系统研究,糖蜜中可发酵糖75％的转化是在1号反应器中完成的。     

Construction of flavocytochrome <Emphasis Type="Italic">b</Emphasis><Subscript>2</Subscript>-overproducing strains of the thermotolerant methylotrophic yeast <Emphasis Type="Italic">Hansenula polymorpha</Emphasis> (<Emphasis Type="Italic">Pichia angusta</Emphasis>)

L-Lactate cytochrome c oxidoreductase (flavocytochrome b ₂, FC b ₂) from the thermotolerant methylotrophic yeast Hansenula polymorpha (Pichia angusta) is, unlike the enzyme form baker’s yeast, a thermostable enzyme potentially important for bioanalytical technologies for highly selective assays of L-lactate in biological fluids and foods. This paper describes the construction of flavocytochrome b ₂ producers with over-expression of the H. polymorpha CYB2 gene, encoding FC b ₂. The HpCYB2 gene under the control of the strong H. polymorpha alcohol oxidase promoter in a plasmid for multicopy integration was transformed into the recipient strain H. polymorpha C-105 (grc1 catX), impaired in glucose repression and devoid of catalase activity. A method was developed for preliminary screening of the transformants with increased FC b ₂ activity in permeabilized yeast cells. The optimal cultivation conditions providing for the maximal yield of the target enzyme were found. The constructed strain is a promising FC b ₂ producer characterized by a sixfold increased (to 3 μmol min^?1 mg^?1 protein in cell-free extract) activity of the enzyme.     

The amino‐terminal tail of Hxt11 confers membrane stability to the Hxt2 sugar transporter and improves xylose fermentation in the presence of acetic acid

Hxt2 is a glucose repressed, high affinity glucose transporter of the yeast Saccharomyces cerevisiae and is subjected to high glucose induced degradation. Hxt11 is a sugar transporter that is stably expressed at the membrane irrespective the sugar concentration. To transfer this property to Hxt2, the N‐terminal tail of Hxt2 was replaced by the corresponding region of Hxt11 yielding a chimeric Hxt11/2 transporter. This resulted in the stable expression of Hxt2 at the membrane and improved the growth on 8% d ‐glucose and 4% d ‐xylose. Mutation of N361 of Hxt11/2 into threonine reversed the specificity for d ‐xylose over d ‐glucose with high d ‐xylose transport rates. This mutant supported efficient sugar fermentation of both d ‐glucose and d ‐xylose at industrially relevant sugar concentrations even in the presence of the inhibitor acetic acid which is normally present in lignocellulosic hydrolysates. Biotechnol. Bioeng. 2017;114: 1937–1945. © 2017 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.     

Relationship between molecular and enological features of Patagonian wine yeasts: relevance in selection protocols

Summary In this paper, a study of the relationship between genetic patterns, obtained by the combination of mtDNA-RFLP and PCR-amplified inter-δ sequence DNA polymorphism analysis, and relevant enological phenotypic data (fermentative power, specific productivity, volatile and total acidity) was carried out on Argentinean Saccharomyces cerevisiae isolates from north Patagonia. The use of a powerful statistical tool, Generalized Procrustes analysis, allowed us to weigh the relationship for each isolate in particular, denoting a good enough degree of agreement between molecular and physiological data for most of the population analysed. The inclusion of a physiological feature, as the killer sensitivity biotype, within identification methods resulted in a higher degree of discrimination among isolates and in better correlation between both characterizations. The combined use of methods based on molecular polymorphisms and killer biotype could be applied so as not to miss any isolate with differential enological properties in selection protocols.     

PML coiled-coil结构域与RAN结合蛋白9相互作用的验证

目的验证前髓细胞性白血病的卷曲螺旋结构域（PML—C）和RAN结合蛋白9（RANBP9）之间的相互作用。方法构建分别表达诱饵蛋白PML—C和靶蛋白RANBP9的载体pGBKT7-PML-C和pACT2-RANBP9,然后转人酵母AH109,培养3～5d后对其是否有胞内相互作用进行检测。将目的片断PML-C和RANBP9再次构建于真核生物表达载体pCMV—HA和pCMV—myc里,然后共转染人胚肾293细胞里（HEK293）,最后对其是否有体外相互作用通过免疫共沉淀和免疫印迹进行分析。结果在共转化了质粒pGBKT7-PML—C和pACT2-RANBP9的AH109酵母平板里观察到蓝色菌落生长。用抗HA多克隆抗体对共转染过重组质粒的HEK293细胞的蛋白提取物进行免疫共沉淀,再用抗myc单克隆抗体作为一抗进行免疫印迹,最终检测出融合蛋白myc—RANBP9条带。结论酵母双杂交实验验证了PML—C和RANBP9之间存在胞内相互作用,同时免疫共沉淀实验也从体外验证了它们之间的相互作用。     

环指蛋白RING1与A型核纤层蛋白的细胞内相互作用

环指蛋白RING1能结合DNA并抑制基因的转录.采用酵母双杂交方法从人骨骼肌文库中筛选出了与A型核纤层蛋白(lamin A)结合的RING1蛋白,回复杂交酵母能在缺陷培养基上生长.RING1与绿色荧光蛋白融合载体转染HEK293细胞,激光共聚焦显微观察发现RING1能与带红色荧光蛋白的lamin A蛋白在细胞核周围共定位.免疫共沉淀结果证明RING1与lamin A能够相互作用.结果证明了一个新的lamin A结合蛋白,为揭示lamin A影响基因表达乃至细胞衰老提供了依据.     

虹鳟生长激素cDNA在酵母中的表达

采用聚合酶链式反应（ Ｐ Ｃ Ｒ） 技术对虹鳟生长激素ｃ Ｄ Ｎ Ａ 进行改造。将改造后的基因克隆到含酵母 Ｐ Ｇ Ｋ 启动子的大肠杆菌酵母穿梭质粒ｐ Ｍ Ａ９１ ,转化酿酒酵母 Ｙ３３ ,构建表达鱼生长激素的酵母工程菌 Ｙ３３（ｐ Ｍ Ａｒ Ｇ Ｈ１６） ,并在酵母中获得表达,表达量约占细胞可溶性蛋白总量的３ ％ 。表达产物作为饲料添加剂投喂罗非鱼,具有明显的促进生长作用