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1.
实验将自絮凝颗粒酵母培养于同时添加脂肪酸 (0.6mmol/L)和酒精 (6 %~ 9% ,V/V)条件下以考察其细胞膜磷脂脂肪酸组成的变化。与单独添加棕榈酸相比 ,同时添加酒精引起细胞膜磷脂棕榈酸含量明显增加 ,伴随 9十四碳烯酸、棕榈油酸和油酸含量明显减少 ;与单独添加亚油酸相比 ,同时添加酒精未引起细胞膜磷脂亚油酸含量明显变化 ,但引起油酸含量明显增加 ,伴随 9 十四碳烯酸、棕榈油酸和棕榈酸含量减少 ;与单独添加亚麻酸相比 ,同时添加酒精引起细胞膜磷脂亚麻酸含量减少 ,伴随油酸含量显著增加 ,同时 9 十四碳烯酸、棕榈油酸和棕榈酸含量减少。存活率实验证实 ,上述变化是菌体对酒精刺激的适应性响应 ,因为 ,与培养于仅添加脂肪酸条件下的菌体相比 ,培养于同时添加酒精条件下的菌体耐酒精能力明显提高。研究表明 ,棕榈酸和油酸都可通过加强细胞膜渗透屏障而提高菌体的耐酒精能力 ,这是饱和脂肪酸 (SFA)与不饱和脂肪酸 (UFA)可提高同一菌株耐酒精能力的新的实验现象 ,揭示UFA与SFA在影响酵母菌耐酒精能力的机制上存在共同的作用方式  相似文献   

2.
实验显示,一种氨基酸混合液(含异亮氨酸、甲硫氨酸和苯丙氨酸,添加浓度分别为1.0、0.5和2.0g/L)能显著提高自絮凝酵母——粟酒裂殖酵母和酿酒酵母融合株SPSC的耐酒精能力。实验将菌体分别培养于添加(试验组)和未添加(对照组)该氨基酸混合液的条件下,然后收集菌体进行酒精(20%,V/V)冲击试验(30℃,9h),结果,试验组的菌体尚有一半以上的存活细胞,而对照组的菌体全部死亡。通过对试验组和对照组的菌体细胞膜蛋白质氨基酸组成分析发现,试验组的菌体耐酒精能力提高与所添加氨基酸组入菌体的细胞膜密切相关。以DPH为荧光探针的细胞膜流动性测定分析进一步揭示,氨基酸组入菌体的细胞膜后,细胞膜能有效抵抗高浓度酒精冲击诱发的膜流动性的提高,从而维持膜的稳定。因此,实验首次揭示膜蛋白氨基酸组成可通过改变膜流动性而影响酵母菌的耐酒精能力。  相似文献   

3.
研究揭示细胞膜磷脂脂肪酸组成与质膜ATP酶在酵母菌耐酒精中的一种新颖关系。实验表明 ,细胞膜磷脂脂肪酸组成特点对生长于未添加酒精条件下的自絮凝颗粒酵母质膜ATP酶活性没有影响 ,但却明显影响生长于添加酒精 (1 %~ 10 % ,V/V)条件下的菌体质膜ATP酶对酒精激活的敏感性 :预培养于添加 0.6mmol L棕榈酸、亚油酸、或亚麻酸条件下的菌体的质膜ATP酶的最大激活水平分别为各自酶的基态水平 (未激活 )的 3.6、1.5和 1.2倍 ,而对照组 (预培养于未添加脂肪酸条件下的菌体 )的相应值为2.3倍 ,酶产生上述最大激活水平时的酒精浓度分别为 7%、6 %、6 %、和 7% (V/V)。酶激活后米氏常数Km 、最适pH和对钒酸钠 (质膜ATP酶特异性抑制剂 )的敏感性等性质不变 ,但最大反应速度vmax明显增加。实验表明 ,细胞膜磷脂脂肪酸组成特点对提高菌体的耐酒精能力越有利 ,则其质膜ATP酶被酒精激活的幅度越大 ,说明菌体耐酒精能力的提高与其质膜ATP酶对酒精激活的敏感性的增加密切相关。细胞膜磷脂脂肪酸组成会影响酵母菌质膜ATP酶对酒精激活的敏感性是观察到的新的实验现象.  相似文献   

4.
酿酒酵母属(S. cereviae)变异株和粟酒裂殖酵母属(S. pombe)变异株进行属间原生质体融合得到融合株SPSC,该融合株比S. cereviae具有强的自身絮凝能力。以葡萄糖浓度150g/L的底物在30~44℃的温度范围内进行摇瓶厌氧发酵,获得最佳温度范围为34~38℃,最高发酵温度为40℃。在有效容积2.35L悬浮床反应器中,在pH值3.0~5.0范围内进行连续发酵,获得最适发酵pH为3.5~4.5。  相似文献   

5.
研究揭示细胞膜磷脂脂肪酸组成与酵母菌耐酒精能力的一种新颖关系及其机制。分别培养于添加 0 6mmol L棕榈酸、亚油酸或亚麻酸不同条件下的自絮凝颗粒酵母 ,其细胞膜富含各自所添加的脂肪酸。细胞膜富含棕榈酸、亚油酸或亚麻酸的三种菌体于 30℃经 2 0 %(v v)酒精冲击 6h的存活率分别为 5 2 %、1 8%和 0。通过考察三种菌体于 30℃在 1 5 %(v v)酒精冲击下的细胞膜透性发现 ,细胞膜富含棕榈酸的菌体的胞外核苷酸平衡浓度分别仅为细胞膜富含亚油酸或亚麻酸菌体的 48%和 32 %,其细胞膜透性系数 (P′)分别仅为后者的 37%和 2 0 %,且三者的胞外核苷酸浓度和P′由小到大的排列顺序均与它们的存活率由高到低的排列顺序完全一致。因此 ,细胞膜富含棕榈酸的菌体具有较强的耐酒精能力是与其在高浓度酒精冲击下可维持较低的细胞膜透性密切相关的 的。  相似文献   

6.
絮凝颗粒酵母均匀悬浮体系生长动力学的研究   总被引:3,自引:0,他引:3  
利用粟酒裂殖酵母(Schizosaccharomyces pombe)变异株自身絮凝形成的颗粒,作为细胞固定化方法。以双酶法制备的淀粉耱化液为底物,在有效容积2.35L的小型悬浮床生物反应器中连续生产酒精。研究了微量供氧条件下该絮凝颗粒酵母均匀悬浮体系的生长动力学,获得了描述其生长规律的模型方程。  相似文献   

7.
酿酒酵母属(S. cereviae)变异株和粟酒裂殖酵母属(S. pombe)变异株进行属间原生质体融合得到融合株SPSC,该融合株比S. cereviae具有强的自身絮凝能力。以葡萄糖浓度150g/L的底物在30~44℃的温度范围内进行摇瓶厌氧发酵,获得最佳温度范围为34~38℃,最高发酵温度为40℃。在有效容积2.35L悬浮床反应器中,在pH值3.0~5.0范围内进行连续发酵,获得最适发酵pH为3.5~4.5。  相似文献   

8.
絮凝酵母SPSC01为酿酒酵母Saccharomyces cerevisiae和粟酒裂殖酵母Schizosaccharomyces pombe的融合菌株,用其吸附水溶液中的重金属Cr(VI),可以大大降低生物吸附的固液分离成本。为了探讨SPSC01菌体絮凝蛋白对Cr(VI)还原吸附的影响,对SPSC01与其亲本菌株的吸附行为进行了比较。结果表明,SPSC01和其具有絮凝性状的亲本S.pombe的Cr(VI)去除速率基本同步,远优于无絮凝性状的亲本S.cerevisiae;达到吸附平衡时,S.pombe、SPSC01和S.cerevisiae对总Cr去除率分别达68.8%、48.6%和37.5%;从而证明了絮凝有利于Cr(VI)的还原、吸附,絮凝蛋白在Cr(VI)的还原吸附过程中起促进作用。通过化学屏蔽方法和傅立叶变换红外光谱(FTIR)分析,对SPSC01菌体表面吸附Cr(VI)的机理进行了研究,结果表明SPSC01菌体表面吸附Cr(VI)起主要作用的基团是氨基、羧基和酰胺基。  相似文献   

9.
研究揭示细胞膜磷脂脂肪酸组成与质膜ATP酶在酵母菌耐酒精中的一种新颖关系。实验表明,细胞膜磷脂脂肪酸组成特点对生长于未添加酒精条件下的自絮凝颗粒酵母质膜ATP酶活性没有影响,但却明显影响生长于添加酒精(1%~10%,V/V)条件下的菌体质膜ATP酶对酒精激活的敏感性:预培养于添加0.6mmol/L棕榈酸、亚油酸、或亚麻酸条件下的菌体的质膜ATP酶的最大激活水平分别为各自酶的基态水平(未激活)的3.6、1.5和1.2倍,而对照组(预培养于未添加脂肪酸条件下的菌体)的相应值为2.3倍,酶产生上述最大激活水平时的酒精浓度分别为7%、6%、6%、和7%(V/V)。酶激活后米氏常数Km、最适pH和对钒酸钠(质膜ATP酶特异性抑制剂)的敏感性等性质不变,但最大反应速度υmax明显增加。实验表明,细胞膜磷脂脂肪酸组成特点对提高菌体的耐酒精能力越有利,则其质膜ATP酶被酒精激活的幅度越大,说明菌体耐酒精能力的提高与其质膜ATP酶对酒精激活的敏感性的增加密切相关。细胞膜磷脂脂肪酸组成会影响酵母菌质膜ATP酶对酒精激活的敏感性是观察到的新的实验现象。  相似文献   

10.
絮凝酵母SPSC01为酿酒酵母Saccharomyces cerevisiae和粟酒裂殖酵母Schizosaccharomyces pombe的融合菌株,用其吸附水溶液中的重金属Cr(VI),可以大大降低生物吸附的固液分离成本。为了探讨SPSC01菌体絮凝蛋白对Cr(VI) 还原吸附的影响,对SPSC01与其亲本菌株的吸附行为进行了比较。结果表明,SPSC01和其具有絮凝性状的亲本S. pombe的Cr(VI) 去除速率基本同步,远优于无絮凝性状的亲本S. cerevisiae;达到吸附平衡时,S. pombe、SPSC01和S. cerevisiae对总Cr去除率分别达68.8%、48.6%和37.5%;从而证明了絮凝有利于Cr(VI) 的还原、吸附,絮凝蛋白在Cr(VI) 的还原吸附过程中起促进作用。通过化学屏蔽方法和傅立叶变换红外光谱 (FTIR) 分析,对SPSC01菌体表面吸附Cr(VI) 的机理进行了研究,结果表明SPSC01菌体表面吸附Cr(VI) 起主要作用的基团是氨基、羧基和酰胺基。  相似文献   

11.
Lei J  Zhao X  Ge X  Bai F 《Journal of biotechnology》2007,131(3):270-275
The ethanol tolerance of a self-flocculating yeast strain SPSC01 was investigated in an oxygen-limited fed-batch bioreactor. Employing Focused Beam Reflectance Measurement (FBRM) on-line monitoring system, four yeast floc populations with the average size ranging from 100 to 400mum were obtained. It was found that ethanol tolerance increased with the increasing floc size in the 100, 200, and 300mum floc populations, while increasing the average floc size further to 400mum resulted in lower ethanol tolerance. Examination of the membrane composition of different floc populations revealed that the plasma membrane composition of the floc populations was significantly different in the contents of ergosterol, phosphatidylinositol, as well as phospholipid palmitoleic acid. What's more, the plasma membrane of more ethanol tolerant floc population was less permeable when subjected to 15% (v/v) ethanol shock treatment, and the plasma membrane ATPase activities were higher in the floc populations with higher ethanol tolerance. These results indicate that the average size distribution of the floc populations exerted great influence on the physiological status of yeast cells during the ethanol production process, leading to the changes in plasma membrane composition that contributed to improved ethanol tolerance in self-flocculating yeast SPSC01.  相似文献   

12.
Yeast flocculation has been used in the brewing industry to facilitate biomass recovery for a long time, and thus its mechanism of yeast flocculation has been intensively studied. However, the application of flocculating yeast in ethanol production garnered attention mainly in the 1980s and 1990s. In this article, updated research progress in the molecular mechanism of yeast flocculation and the impact of environmental conditions on yeast flocculation are reviewed. Construction of flocculating yeast strains by genetic approach and utilization of yeast flocculation for ethanol production from various feedstocks were presented. The concept of self-immobilized yeast cells through their flocculation is revisited through a case study of continuous ethanol fermentation with the flocculating yeast SPSC01, and their technical and economic advantages are highlighted by comparing with yeast cells immobilized with supporting materials and regular free yeast cells as well. Taking the flocculating yeast SPSC01 as an example, the ethanol tolerance of the flocculating yeast was also discussed.  相似文献   

13.
In this investigation, we examined the effects of different unsaturated fatty acid compositions of Saccharomyces cerevisiae on the growth-inhibiting effects of ethanol. The unsaturated fatty acid (UFA) composition of S. cerevisiae is relatively simple, consisting almost exclusively of the mono-UFAs palmitoleic acid (Delta(9)Z-C(16:1)) and oleic acid (Delta(9)Z-C(18:1)), with the former predominating. Both UFAs are formed in S. cerevisiae by the oxygen- and NADH-dependent desaturation of palmitic acid (C(16:0)) and stearic acid (C(18:0)), respectively, catalyzed by a single integral membrane desaturase encoded by the OLE1 gene. We systematically altered the UFA composition of yeast cells in a uniform genetic background (i) by genetic complementation of a desaturase-deficient ole1 knockout strain with cDNA expression constructs encoding insect desaturases with distinct regioselectivities (i.e., Delta(9) and Delta(11)) and substrate chain-length preferences (i.e., C(16:0) and C(18:0)); and, (ii) by supplementation of the same strain with synthetic mono-UFAs. Both experimental approaches demonstrated that oleic acid is the most efficacious UFA in overcoming the toxic effects of ethanol in growing yeast cells. Furthermore, the only other UFA tested that conferred a nominal degree of ethanol tolerance is cis-vaccenic acid (Delta(11)Z-C(18:1)), whereas neither Delta(11)Z-C(16:1) nor palmitoleic acid (Delta(9)Z-C(16:1)) conferred any ethanol tolerance. We also showed that the most ethanol-tolerant transformant, which expresses the insect desaturase TniNPVE, produces twice as much oleic acid as palmitoleic acid in the absence of ethanol and undergoes a fourfold increase in the ratio of oleic acid to palmitoleic acid in response to exposure to 5% ethanol. These findings are consistent with the hypothesis that ethanol tolerance in yeast results from incorporation of oleic acid into lipid membranes, effecting a compensatory decrease in membrane fluidity that counteracts the fluidizing effects of ethanol.  相似文献   

14.
【目的】利用转录组测序研究硫酸锌添加提高絮凝酿酒酵母SPSC01乙酸胁迫耐性的分子机理。【方法】在10.0 g/L乙酸胁迫条件下,添加0.03 g/L硫酸锌,取对数期酿酒酵母细胞,与不添加硫酸锌的对照组细胞进行比较转录组分析。【结果】添加硫酸锌的实验组与对照组相比较,50个基因转录水平上调,162个基因转录水平下调,这些转录水平变化明显的基因涉及糖代谢、甲硫氨酸合成、维生素合成等多条代谢途径,此外,转录水平变化的基因还包括抗氧化酶基因等关键胁迫响应基因。【结论】硫酸锌添加可改变酿酒酵母全局基因转录水平,提高抗氧化酶及其他胁迫耐性相关基因的表达,影响细胞氧化还原平衡和能量代谢,通过对多基因转录的调控提高酿酒酵母乙酸耐受性。  相似文献   

15.
Abstract The lipid composition of a strain of each of two yeasts, Saccharomyces csrevisiae and Kloeckera apiculata , with different ethanol tolerances, was determined for cells grown with or without added ethanol. An increase in the proportion of ergosterol, unsaturated fatty acid levels and the maintenance of phospholipid biosynthesis seemed to be responsible for ethanol tolerance. The association of ethanol tolerance of yeast cells with plasma membrane fluidity, measured by fluorescence anisotropy, is discussed. We propose that an increase in plasma membrane fluidity may be correlated with a decrease in the sterol: phospholipid and sterol: protein ratios and an increase in unsaturation index.  相似文献   

16.
Yeast flocculation is an important trait in the brewing industry as well as in ethanol production, through which biomass can be recovered by cost-effective sedimentation. However, mass transfer limitation may affect yeast growth and ethanol fermentation if the flocculation occurs earlier before fermentation is completed. In this article, a novel type of cell-cell flocculation induced by trehalose-6-phosphate synthase 1 (TPS1) promoter was presented. The linear cassette HO-P(TPS1)-FLO1(SPSC01)-KanMX4-HO was constructed to transform the non-flocculating industrial yeast S. cerevisiae 4126 by chromosome integration to obtain a new flocculating yeast strain, ZLH01, whose flocculation was induced by ethanol produced during fermentation. The experimental results illustrated that flocculation of ZLH01 was triggered by 3% (v/v) ethanol and enhanced as ethanol concentration increased till complete flocculation was achieved at ethanol concentration of 8% (v/v). Real time PCR analysis confirmed that the expression of FLO1(SPSC01) was dependent on ethanol concentration. The growth and ethanol fermentation of ZLH01 were improved significantly, compared with the constitutive flocculating yeast BHL01 engineered with the same FLO gene but directed by the constitutive 3-phosphoglycerate kinase promoter PGK1, particularly under high temperature conditions. These characteristics make the engineered yeast more suitable for ethanol production from industrial substrates under high gravity and temperature conditions. In addition, this strategy offers advantage in inducing differential expression of other genes for metabolic engineering applications of S. cerevisiae.  相似文献   

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