碳酸钙促进丙酮酸发酵过程中α-酮戊二酸的形成

在多重维生素营养缺陷型菌株光滑球拟酵母CCTCC M202019发酵生产丙酮酸的摇瓶和发酵罐实验中发现,CaCO₃的添加对发酵液中α-酮戊二酸（α-KG）的积累有重要影响。在维生素浓度不变且供氧充分的前提下,延迟CaCO₃添加时间可明显抑制α-KG的产生,并提高丙酮酸与α-KG的碳摩尔比(C_PYR/C_αKG);而增加培养基中的CaCO₃浓度会导致αKG积累的增加。用不同物质调节发酵液中pH的实验证实：在丙酮酸发酵过程中, Ca²⁺对αKG的积累起主要作用,CO₃^2-起辅助作用,两者对α-KG的积累具有协同效应。维持培养基中CaCO₃浓度不变,改变培养基中硫胺素的浓度,对αKG的积累,特别是对C_PYR/C_α-KG值没有影响;而增加培养基中生物素的浓度,则导致αKG的浓度不断上升且C_PYR/C_α-KG值不断下降。当有Ca2+存在时,胞内丙酮酸羧化酶的活性最高可提高40%,而丙酮酸脱氢酶系的活性没有明显变化。结果表明,丙酮酸发酵过程中α-KG的形成是由于CaCO₃促进了丙酮酸羧化反应,其中Ca²⁺可显著提高丙酮酸羧化酶的活性,而CO₃^2-则有可能作为丙酮酸羧化反应的底物。     

康氏木霉纤维素酶系中Cx酶多分子型的分离纯化及某些性质

从康氐木霉(Trichoderma kkoningii) 白色变异株 AS 3．4001的粗酶制剂中,获得了纤维素酶系中的一组C_x酶(C_xt C_xz C_z3 C_z4)。分离步骤包括Sephadcx G-75凝胶过滤,DEAE-Sephadex A-50离子交换层析,ConA-Sepharnse亲合层析,SE—Sephadcx C-50离子交换层析及聚丙烯酰胺凝胶电泳。C_xt 与C_xt 的分子量不同而所带电荷相同,它们的分子量各自为44,500和34,000。C_xz—C_x4 的分子量相同而所带电荷不同。纯化的C_xt—C_z4“经聚丙烯酰胺凝胶电泳鉴定为单带。比较它们对羧甲基纤维素钠(CMC—Na)的糖化力及液化力表明在作用方式的随机性上C_xz>C_z3>C_z1>C_x4。     

What is the usual internal carbon dioxide concentration in C<Subscript>4</Subscript> species under midday field conditions?

The carbon dioxide concentrating system in C₄ photosynthesis allows high net photosynthetic rates (P _N) at low internal carbon dioxide concentrations (C _i), permitting higher P _N relative to stomatal conductance (g _s) than in C₃ plants. This relation would be reflected in the ratio of C _i to external ambient (C _a) carbon dioxide concentration, which is often given as 0.3 or 0.4 for C₄ plants. For a C _a of 360 μmol mol⁻¹ that would mean a C _i about 110–140 μmol mol⁻¹. Our field observations made near midday on three weedy C₄ species, Amaranthus retroflexus, Echinochloa crus-galli, and Setaria faberi, and the C₄ crop Sorghum bicolor indicated mean values of C _i of 183–212 μ mol mol⁻¹ at C _a = 360 μmol mol⁻¹. Measurements in two other C₄ crop species grown with three levels of N fertilizer indicated that while midday values of C _i at high photon flux were higher at limiting N, even at high nitrogen C _i averaged 212 and 196 μmol mol⁻¹ for Amaranthus hypochondriacus and Zea mays, respectively. In these two crops midday C _i decreased with increasing leaf to air water vapor pressure difference. Averaged over all measurement days, the mean C _i across all C₄ species was 198 μmol mol⁻¹, for a C _i/C _a ratio of 0.55. Prior measurements on four herbaceous C₃ species using the same instrument indicated an average C _i/C _a ratio of 0.69. Hence midday C _i values in C ₄ species under field conditions may often be considerably higher and more similar to those of C₃ species than expected from measurements made on plants in controlled environments. Reducing g _s in C₄ crops at low water vapor pressure differences could potentially improve their water use efficiency without decreasing P _N.     

Smac/DIABLO在过氧化氢所致C2C12肌原细胞凋亡中的作用

为探讨Smac/DIABLO在过氧化氢(H₂O₂）所致C₂C₁₂肌原细胞凋亡中的作用,采用Hoechst 33258染色,观察H₂O₂ (0.5 mmol/L)处理C₂C₁₂肌原细胞不同时间后,细胞核形态学改变并计算凋亡核百分率,DNA抽提及琼脂糖电泳观察凋亡特征性梯状带,利用细胞成分分离后蛋白质印迹分析H₂O₂是否导致Smac/DIABLO从线粒体释放,采用Caspase检测试剂盒及蛋白质印迹分析Caspase-3和Caspase-9的活化,转染Smac/DIABLO基因,观察Smac/DIABLO过表达对H₂O₂所致的C₂C₁₂肌原细胞凋亡的影响.结果表明:H₂O₂处理1 h后,Smac/DIABLO从C₂C₁₂肌原细胞线粒体释放入胞浆,2 h更明显;H₂O₂处理4 h后,Caspase-3和Caspase-9活化,12 h达高峰;H₂O₂处理24 h后,C₂C₁₂肌原细胞显示特征性的凋亡形态改变,凋亡核百分率明显升高,DNA电泳出现明显“梯状”条带.与单纯过氧化氢损伤组相比,Smac/DIABLO高表达的C₂C₁₂肌原细胞经过氧化氢损伤组的Caspase-3和Caspase-9的活化、凋亡核百分率的升高、“梯状”条带的出现均更明显.结果表明,H₂O₂可导致Smac/DIABLO从C₂C₁₂肌原细胞线粒体释放,促进Caspase-9和Caspase-3的活化而促进细胞凋亡的发生.     

西藏冬虫夏草无性型的分子生物学研究*

从西藏采集的冬虫夏草子实体中分离出 3个菌株C₃、C₄、C₅,通过培养特征的研究以及采用分子生物学方法 ,以rDNAITS区为分子指标 ,与冬虫夏草 [Cordycepssinensis(Berk .)Sacc.) ]的有性世代及中国被毛孢 (HirsutellasinensisLiu ,Guo,Yu&Zeng)进行比较分析 ,发现C4的培养特征与C₃、C₅ 完全不同 ,其序列与冬虫夏草的相似率为     

云贵高原喀斯特坡耕地土壤微生物量 C、N、P空间分布

土壤微生物是地球生物演化进程中的先锋种类,具有重要的生态修复功能,但空间分布格局是否存在的争议很大。以云贵高原典型喀斯特坡耕地为对象,基于网格法取样,用经典统计学和地统计学综合分析了土壤微生物生物量的空间变异特征。结果表明,云贵高原喀斯特坡耕地土壤微生物生物量碳(C_mic)、磷(P_mic)以及碳氮比(C_mic/N_mic)适宜,氮(N_mic)的含量较低,变异均很大,空间自相关性明显,最佳拟合模型均为指数模型。块金值C₀较小(0.0016-0.0087),C₀/(C₀+C)均<25%(2.6%-10.2%),变程a较短(22.2-51.0 m),其强烈的空间变异主要由结构性变异引起。Kriging等值线图表明,C_mic、N_mic和C_mic/N_mic的高值区分布在坡的中上部,P_mic的高值区则在坡的中下部和坡脚。云贵高原喀斯特坡耕地土壤微生物不仅存在着小尺度的空间分布格局,而且不同土壤微生物属性的空间分布不同。     

C60对体外培养癌细胞的光致作用研究

研究结果表明用C₆₀-磷脂酰胆碱与体外培养的HeLa细胞融合后,以4 000 lx光强度激发C₆₀(C₆₀浓度20 mg／L)对HeLa细胞具有显著的光致杀伤作用.生化测定证实光激发C₆₀导致膜蛋白巯基含量减少、膜脂过氧化、丙二醛含量增高,SDS-PAGE证明膜蛋白交联,荧光偏振显示膜流动性降低,电镜超微膜结构破坏,经MTT法检测,大部分细胞死亡.C₆₀的强烈光致作用证实了Arbogast等认为光激发C₆₀可产生单线态氧的观态.     

带状采伐后不同时期毛竹林恢复和土壤养分特征

为研究带状采伐毛竹林对竹林恢复和土壤养分的影响,以福建省建瓯市毛竹林为研究对象,设置5 m(处理1,C₁)和7 m(处理2,C₂)2种采伐宽度,以不进行采伐处理为对照(CK),研究采伐后当年新鞭生长期(I时期,2018年10月)和采伐后次年新立竹生长期(Ⅱ时期,2019年6月)新竹生长相关指标和土壤养分变化特征。结果表明：(1)C₁、C₂、CK处理的出笋数量分别为2 227、2 650和1 955株·hm^-2,且C₂与C₁、CK差异显著(P<0.05);退笋率各处理之间无显著差异(P>0.05),表现为C₁2。(2)2种采伐处理的胸径(DBH)显著低于CK,表现为CK>C₂>C₁。(3)采伐后Ⅰ时期和Ⅱ时期相比,土壤全磷(TP)和全钾(TK)含量增加,全氮(TN)和全碳(TC)含量降低;采伐后I时期的C₁、C₂处理中土壤TN、TP、TK和TC含量均大于CK,但采伐后Ⅱ时期的土壤TC含量小于CK。研究认为,带状采伐能提高出笋量和立竹数,增加土壤氮磷钾养分含量;7 m带状采伐竹林恢复状况整体上优于5 m带状采伐。     

早春短命植物鸢尾蒜和准噶尔鸢尾蒜的光合途径

该研究以鸢尾蒜属两种早春短命植物准噶尔鸢尾蒜(Ixiolirion songaricum)和鸢尾蒜(Ixiolirion tataricum)为对象,通过解剖结构、光合参数、稳定碳同位素比值(δ¹³C)和光合关键酶活性分析二者的光合途径。结果发现：(1)显微结构和超微结构显示,两种短命植物的叶脉维管束鞘细胞1层、排列较紧密,鞘细胞内含有较多叶绿体且多离心分布,类似C₄花环结构。(2)准噶尔鸢尾蒜和鸢尾蒜最大净光合速率分别为14.81和15.04 μmol·m^-2·s^-1;两者的CO₂补偿点较低,分别为3.57和2.54 μmol·mol^-1,δ¹³C分别为-25.36±0.55‰和-25.76±1.38‰,光合酶PEPC/Rubisco比值分为别0.244和0.322。(3)最大净光合速率、δ¹³C和PEPC/Rubisco比值均说明两种植物为C₃光合途径,但二者均具有类似C₄的维管束鞘结构、较低的CO₂补偿点和暗呼吸速率,并具有高于部分C₃和C₃ C₄中间型植物的PEPC/Rubisco比值,表明两种短命植物的光合途径并非典型的C₃途径,而是C₃ C₄中间型。     

应用稳定同位素技术分析华北部分地区第三代棉铃虫虫源性质

玉米等C₄植物被认为是华北Bt棉种植区内第三代棉铃虫最重要的天然庇护所,但尚缺乏直接证据。连续2a(2006-2007年)利用杨树把诱集棉铃虫成虫,进行碳稳定同位素比值的测定,并结合棉铃虫成虫捕获时间、虫源的数量比例等,评估C₄植物在华北第三代棉铃虫期间的庇护所功能。结果表明,第三代棉铃虫成虫来源于C₄植物(玉米)的为40.5%-56.8%,与C₃来源的数量上大体相当。但C₄来源的成虫羽化时间比C₃来源的个体明显滞后,呈现出先少后多的特点。结果提示,C₄植物确实是华北第三代棉铃虫重要的庇护所,但存在着时间上与C₃来源的成虫交配不同步而失效的风险;结果建议玉米等天然庇护所作物的种植不仅在面积上要足够,而且播种时间上要充分考虑C₄植物(玉米)来源的敏感棉铃虫个体的发育与C₃植物寄主来源个体的同步性。     

Physiological and Biochemical Characteristics of Immobilized Champagne Yeasts and Their Participation in Champagnizing Processes: A Review

Methods for immobilizing champagne yeasts, physiological and biochemical characteristics of the immobilized cells, and problems of their utilization in the production of quality champagne wines are reviewed. Studies aimed at the development of efficient biocatalysts for champagnizing wines using bottle fermentation (methode champenoise) and tank processing (bulk, or Charmat process) based on the use of immobilized yeast cells are described. Data on the industrial use of such biocatalysts in countries manufacturing champagne wines are presented. Problems and prospects of further research in this field are discussed.     

Physiological and biochemical characteristics of immobilized champagne yeasts and their participation in sparkling processes

Methods for immobilizing champagne yeasts, physiological and biochemical characteristics of the immobilized cells, and problems of their utilization in the production of quality champagne wines are reviewed. Studies aimed at the development of efficient biocatalysts for champaignizing wines using bottle fermentation (method champenoise) and tank processing (bulk, or Charmat process), based on the use of immobilized yeast cells, are described. Data on the industrial use of such biocatalysts in countries manufacturing champagne wines are presented. Problems and prospects of further research in this field are discussed.     

Immobilization of champagne yeasts by inclusion into cryogels of polyvinyl alcohol for preventing cell escape from carrier matrix

Wine champagnizing, a process involving the use of champagne yeasts immobilized by inclusion into cryogels of polyvinyl alcohol, has been studied. Treatment of yeast cells with the autoregulatory factor d1 was proposed as a means of preventing the cell escape from the carrier matrix. Such a treatment inhibited growth and proliferation processes in yeasts cells, without affecting the activity of fermentation; the resulting champagne had the same organoleptic and chemical characteristics as its counterparts obtained using traditional techniques.     

Immobilization of Champagne Yeasts by Inclusion into Cryogels of Polyvinyl Alcohol: Means of Preventing Cell Release from the Carrier Matrix

Wine champagnizing, a process involving the use of champagne yeasts immobilized by inclusion into cryogels of polyvinyl alcohol, has been studied. Treatment of yeast cells with the autoregulatory factor d ₁ was proposed as a means of preventing the cell release from the carrier matrix. Such a treatment inhibited growth and proliferation processes in yeast cells, without affecting the activity of fermentation; the resulting champagne had the same organoleptic and chemical characteristics as its counterparts obtained using conventional techniques.     

Theme 4: Immobilized Cell Technology in Wine Production

Abstract

In spite of its traditional nature, wine making is largely concerned with the progress of biotechnology. High cell density reactors have potential for enology: improved performance of alcoholic and malolactic fermentations, smaller scale fermentation facilities, adaptation to continuous processes. Among the immobilization techniques, cell entrapment in alginate beads seems to be an impressive one. Alcoholic fermentation of wine, malolactic fermentation, bottle fermentation known as “Methode champenoise” and sparkling wine are among the industrial applications. Knowledge of kinetics and physiology in microorganisms in heterogeneous media has expanded in the last few years. The use of immobilized yeast cells for the champagne method would greatly simplify “remuage”. The compared metabolism of entrapped and free cells during the bottle fermentation shows differences, but the final product does not reveal significant sensory disparity. New products can be obtained with more thoroughly controlled conditions.     

Effect of phenolic compounds, ethyl alcohol, and sodium metabisulphite on the lytic activity of phage PL-1 on a Lactobacillus casei S strain

The effect of phenolic compounds, ethyl alcohol, and sodium metabisulphite on the lytic activity of virulent bacteriophage PL-1 on a Lactobacillus casei S strain isolated from a lactic acid beverage fermentation was investigated. Catechin, caffeic, and gallic acids, commercially produced red, white, and champagne tannins, ethyl alcohol, and sodium metabisulphite inhibited plaque formation. Catechin, caffeic, and gallic acids were the most effective inhibitors of plaque formation. Commercially supplied oenocyanin was not effective.     

一株蛹拟青霉(Paecilomyces militaris )深层发酵产物中抗肿瘤活性物质分析

摘要： 【目的】初步搞清一株蛹拟青霉（Paecilomyces militaris ）菌株RCEF0927在发酵罐发酵条件下发酵液的抗中国仓鼠卵巢瘤（CHO）细胞活性及活性成分的具体组成。【方法】用刃天青（Resazurin）法测定样品对CHO细胞的抑制率;用高效液相色谱-高分辨质谱和活性测定联用的方法进行活性成分分析和鉴定。【结果】活性测定结果表明菌株RCEF0927经发酵罐发酵的发酵液具有较强的抗CHO细胞活性;提取实验结果表明抗肿瘤活性物质能较好地被乙酸乙酯提取出来;液相色谱-质谱-活性测定分析表明     

Monitoring gaseous CO2 and ethanol above champagne glasses: flute versus coupe, and the role of temperature

In champagne tasting, gaseous CO(2) and volatile organic compounds progressively invade the headspace above glasses, thus progressively modifying the chemical space perceived by the consumer. Simultaneous quantification of gaseous CO(2) and ethanol was monitored through micro-gas chromatography (μGC), all along the first 15 minutes following pouring, depending on whether a volume of 100 mL of champagne was served into a flute or into a coupe. The concentration of gaseous CO(2) was found to be significantly higher above the flute than above the coupe. Moreover, a recently developed gaseous CO(2) visualization technique based on infrared imaging was performed, thus confirming this tendency. The influence of champagne temperature was also tested. As could have been expected, lowering the temperature of champagne was found to decrease ethanol vapor concentrations in the headspace of a glass. Nevertheless, and quite surprisingly, this temperature decrease had no impact on the level of gaseous CO(2) found above the glass. Those results were discussed on the basis of a multiparameter model which describes fluxes of gaseous CO(2) escaping the liquid phase into the form of bubbles.     

SPE-HPLC法快速检测发酵液中紫杉醇的含量

针对红豆杉内生真菌发酵液中紫杉醇的含量测定进行探讨,以建立快速高效低耗的检测方法.采用C_(18)固相萃取柱对紫杉醇进行吸附,用不同浓度的甲醇-乙酸铵和甲醇分别作为洗脱剂对其进行洗脱,比较两者的洗脱效果,洗脱液用HPLC进行检测;色谱条件为:流动相甲醇(v):水(v):乙腈(v)=20: 45: 35,流速:0.70 ml/min,检测波长:227 nm.结果表明,浓度为80%的甲醇溶液洗脱效果较好,紫杉醇的回收率为87.6%.     

敲除啤酒酵母 fks1 基因对啤酒酵母自溶性能的影响

以啤酒酵母G-03为模板,扩增得到铜抗性基因(cup 1)和β-葡聚糖合成酶基因(fks 1)。将fks 1连接pMD-18T Vector得到重组质粒pTK,重组质粒pTK和cup1经Bgl Ⅱ、Sal Ⅰ酶切后连接得到重组质粒pKC。Bam HI酶切重组质粒pKC得到以fks 1为整合位点包含cup1的基因片段fks 1::cup1。用此片段转化啤酒酵母工业菌株G-03,通过硫酸铜抗性筛选得到一株啤酒酵母工程菌G-03/C。工程菌连续传代10次后依然能在筛选平板上生长,遗传稳定性良好。主酵结束G-03/C和G-03的辛酸和癸酸含量基本相同。25℃诱导自溶20 d,G-03/C的辛酸、癸酸分别下降57.3%、81.8%,自溶性能减弱。G-03/C的死亡率、双乙酰、浊度及TBA均较原菌有所下降。G-03/C与G-03酿制成品啤酒的常规指标没有较大差别,品评结果表明,G-03/C风味更优。