超临界CO2萃取洋葱挥发油工艺研究

本实验采用超临界CO2萃取技术从冻干洋葱粉中萃取挥发油。以洋葱挥发油得率为考察指标,经单因素及正交试验,考察了萃取温度、萃取压力、CO2流量、萃取时间4个因素对超临界CO2流体萃取的影响。结果表明萃取压力20 MPa,萃取温度35℃,CO2流量为14 kg/h的条件下萃取2.5 h为最佳工艺,洋葱挥发油得率达0.53%。     

薄荷油超临界CO2萃取条件的优化和筛选

以样品中的薄荷脑含量为指标,通过单因素和正交实验对影响薄荷(Mentha haplocalyx Briq.)油超临界CO2萃取的因素进行研究,筛选出薄荷油超临界CO2萃取的最佳条件.研究结果表明,影响样品中薄荷脑萃取率的因素从大到小依次为萃取压力、萃取温度、萃取时间、CO2流量.样品中薄荷脑含量最高的超临界CO2萃取条件为萃取压力10 MPa、萃取温度50℃、CO2流量30 L·h-1且萃取时间1.5 h.     

白果油的提取工艺及其化学成分研究

采用正交实验对超临界CO2萃取白果油的工艺条件进行优化,比较超声波提取、索氏提取、超临界CO2萃取3种方式对白果油提取率的大小,用最佳提取方式分析不同品种白果中油脂的含量,并用GC-MS分析其成分.结果表明: (1)3种提取方式对白果油提取率的大小顺序为:超临界CO2萃取(添加夹带剂)＞索氏提取＞超声波提取.(2)超临界CO2萃取白果油的最佳工艺条件为:温度40℃、压力20 MPa、流速15 L/h、时间3 h并添加石油醚夹带剂,不同品种白果干粉中油脂得率为3.6%～7.11%.(3)GC-MS分析表明:从超临界萃取白果油中鉴定出17种化学成分,其中不饱和脂肪酸占85.4%;超声波和索氏提取相似,分别从提取的白果油中鉴定出10种和9种化学成分,其中不饱和脂肪酸含量分别为90.3%和90.1%;不饱和脂肪酸以十六、十八碳的为主.     

硅藻金色奥杜藻色素的HPLC分析与超临界CO2萃取研究

以硅藻金色奥杜藻(Odontella aurita)为实验材料,利用高效液相色谱法分析了其色素组成与含量,采取超临界CO2萃取技术研究了从干藻粉内提取岩藻黄素的条件。结果表明,该藻主要含有岩藻黄素、硅甲藻黄素、β-胡萝卜素、硅藻黄素等类胡萝卜素以及叶绿素a和叶绿素c1,其中岩藻黄素为该藻含量最高的类胡萝卜素。色素的萃取率与压强、温度、夹带剂含量以及萃取时间呈正相关,夹带剂含量对萃取率影响最大,CO2流速的影响最小;与有机溶剂法相比,超临界CO2萃取岩藻黄素效率略低,而更利于岩藻黄素的选择性萃取及分离提纯;岩藻黄素的SFE-CO2适宜条件为压强400 bar、温度50℃、CO2流速0.2 L/min、夹带剂比例10%、萃取时间2～3 h。     

超临界CO2提取甘草地上部分总黄酮

采用单因素试验对甘草地上部分(茎叶)的超临界CO2提取工艺进行了研究。实验考察了压力、萃取时间、温度及CO2流量对甘草地上部分总黄酮提取率的影响,以总黄酮提取率和含量为指标,系统的研究了超临界二氧化碳萃取法提取甘草地上部分总黄酮的提取效果。得出的最佳工艺参数为:采用40-60目原料,80%乙醇为夹带剂,萃取时间:1.5 h;萃取压力:30.0 MPa;萃取温度:50℃;CO2流量:10 kg.h-1;分离压力:5.8 MPa;分离温度:40℃。实验结果表明超临界二氧化碳萃取甘草总黄酮的提取率2.09%,含量5.42%,工艺具有提取率高,纯度高的特点,为规模化生产甘草总黄酮的提取提供了研究基础。     

超临界CO_2流体萃取苦丁茶多糖的工艺优化

为了探讨超临界CO_2萃取广西苦丁茶中多糖的工艺条件,该研究采用超临界CO_2流体萃取技术分离苦丁茶多糖,利用苯酚-硫酸法对苦丁茶多糖含量进行测定,并考察不同萃取温度(35、40、45、50、55、60℃)、萃取压力(20、25、30、35、40、45、50 MPa)、萃取时间(30、60、90、120、150 min)、夹带剂(甲醇、95%甲醇、50%甲醇、无水乙醇、95%乙醇、50%乙醇)以及夹带剂(95%乙醇)用量(2.0、2.5、3、3.5、4.0、4.5、5.0 mL·min~(-1))对多糖得率的影响,通过设计正交实验方案,对超临界CO_2萃取广西苦丁茶多糖的提取工艺进行优化。结果表明:通过单因素和正交实验考察了苦丁茶多糖提取的主要影响因素,得到的最佳萃取工艺条件为萃取温度50℃,萃取压力40 MPa,夹带剂流量3.5 mL·min~(-1),萃取时间150 min;采用苯酚-硫酸法对苦丁茶多糖含量进行测定。在最优萃取条件下得到的苦丁茶多糖的提取率为7.05%。由此可知,采用超临界CO_2流体萃取,具有提取温度低、萃取率高、萃取周期短、低耗以及污染小等优点,适用于苦丁茶多糖的提取。     

灰树花粗多糖的提取方法比较及其分离纯化

比较了几种常规粗多糖提取法与超临界CO2提取法对灰树花粗多糖提取的差异,同时对灰树花粗多糖进行脱蛋白、脱色研究。试验结果表明:热水浸提法所得灰树花粗多糖得率最高,为28.1%,而超临界CO2提取法所得粗多糖纯度最高,为39.3%。聚酰胺法脱蛋白及脱色效果较好,蛋白脱除率及脱色率分别为69.9%、61.9%,且多糖吸附率较低。将超临界CO2法提取的灰树花粗多糖采用聚酰胺法脱蛋白及脱色后,多糖得率为79.3%,纯度为50.1%。因此聚酰胺法是分离纯化灰树花粗多糖的一种简便、高效的方法。     

省沽油种子超临界CO2 萃取物中角鲨烯和维生素E的GC-MS分析

用气相色谱-质谱联用(GC-MS)方法对超临界CO2萃取出的省沽油种子油中角鲨烯和维生素E进行分析。省沽油种子油的萃取采用超临界CO2萃取技术,压力25MPa,温度50℃,时间2h;种子油用GC-MS方法分析,以正三十二烷为内标,用内标标准曲线法同时测定省沽油种子油中角鲨烯和维生素E的含量。结果表明:超临界CO2萃取种子油的收率为28.83%;所建立的定量分析方法平均回收率在96.0%~99.0%之间,相对标准偏差小于2.8%,线性相关系数大于0.999,方法检测限在2.0~2.5mg/L之间;省沽油种子油中含角鲨烯4.65%,维生素E1.58%。     

不同方法提取的荷叶挥发油化学成分分析

采取超临界CO2萃取和水蒸气蒸馏提取荷叶挥发油,利用GC-MS对它们进行了定性、定量分析。结果表明这两种方法提取荷叶挥发油的化学成分及含量皆有很大差别。超临界CO2萃取的荷叶挥发油更具天然性,超临界CO2萃取法为提取荷叶挥发油的理想方法。     

正交实验优选八角茴香油的超/亚临界CO_2萃取工艺

通过L16(45)正交实验优选最佳的超临界CO2萃取工艺条件,并在此基础上通过二因素随机区组实验优选最佳的亚临界CO2萃取工艺条件,以八角茴香油的得油率作为考查指标,以各实验方案所得茴香油中反式茴香脑的相对含量作为茴香油质量的评价指标.实验结果表明:最佳的亚临界CO2萃取工艺条件为:萃取压力为7 MPa、萃取温度为22 ℃、解析压力Ⅰ为7 MPa、解析温度Ⅰ为30 ℃、解析压力Ⅱ为5 MPa、解析温度Ⅱ为25 ℃,萃取时间为2.0 h,在此条件下八角茴香油的得油率可达12%以上,茴香油中反式茴香脑的相对含量可达91.2178%.采用亚临界CO2萃取,即保持了超临界CO2萃取八角茴香油高品质和天然芳香的优点,又能显著降低设备投资和成产成本,更有利于在生产中推广.     

Characterization of imipenem-resistant Serratia marcescens producing IMP-type and TEM-type beta-lactamases encoded on a single plasmid

To evaluate the roles of blaIMP and blaTEM genes in the resistance of Serratia marcescens against beta-lactams and to find the spreading ways of these genes, 19 clinical isolates of imipenem-resistant Serratia marcescens were analyzed. Six strains bore blaIMP and blaTEM genes on a single plasmid, as confirmed by transferring resistance determinants via conjugation and transformation, and by detecting bla genes with PCR analysis. The six strains showed two different genomic patterns on pulsed-field gel electrophoresis. All the transconjugants and transformants gained high-level resistance to ampicillin, cephalexin, cefoxitin and cefotaxime, and showed a reduced susceptibility to imipenem, but maintained full susceptibility to aztreonam. In addition, the expressions of blaIMP and blaTEM genes were constitutive, either in Serratia marcescens clinical isolates or in their transconjugants and transformants. These findings may explain the rapid spread of the above resistance determinants among Enterobacteriaceae via transmissible plasmids in the clinical setting.     

Low levels of quantitative and molecular genetic differentiation among natural populations of Medicago ciliaris Kroch. (Fabaceae) of different Tunisian eco-geographical origin

In this paper, we analyze the genetic variability in four Tunisian natural populations of Medicago ciliaris using 19 quantitative traits and six polymorphic microsatellite loci. We investigated the amplification transferability of 30 microsatellites developed in the model legume M. truncatula to M. ciliaris. Results revealed that about 56.66% of analyzed markers are valuable genetic markers for M. ciliaris. The most genetic diversity at quantitative traits and microsatellite loci was found to occur within populations (>80%). Low differentiations among populations at quantitative traits Q _ST  = 0.146 and molecular markers F _ST  = 0.18 were found. The majority of measured traits exhibited no significant difference in the level of Q _ST and F _ST . Furthermore, significant correlations established between these traits and eco-geographical factors suggested that natural selection should be invoked to explain the level of phenotypic divergence among populations rather than drift. There was no significant correlation between population differentiation at quantitative traits and molecular markers. Significant spatial genetic structure consistent with models of isolation by distance was detected within all studied populations. The site-of-origin environmental factors explain about 9.07% of total phenotypic genetic variation among populations. The eco-geographical factors that influence more the variation of measured traits among populations are the soil texture and altitude. Nevertheless, there were no consistent pattern of associations between gene diversity (He) and environmental factors.     

On the integration of subthreshold inputs from Perforant Path and Schaffer Collaterals in hippocampal CA1 pyramidal neurons

Using a realistic model of a CA1 hippocampal pyramidal neuron, we make experimentally testable predictions on the roles of the non-specific cation current, I _h , and the A-type Potassium current, I _A , in modulating the temporal window for the integration of the two main excitatory afferent pathways of a CA1 neuron, the Schaffer Collaterals and the Perforant Path. The model shows that the experimentally observed increase in the dendritic density of I _h and I _A could have a major role in constraining the temporal integration window for these inputs, in such a way that a somatic action potential (AP) is elicited only when they are activated with a relative latency consistent with the anatomical arrangement of the hippocampal circuitry.     

Purification of cytochrome a 1 c 1 from Nitrobacter agilis and characterization of nitrite oxidation system of the bacterium

Cytochrome a ₁ c ₁ was highly purified from Nitrobacter agilis. The cytochrome contained heme a and heme c of equimolar amount, and its reduced form showed absorption peaks at 587, 550, 521, 434 and 416 nm. Molecular weight per heme a of the cytochrome was estimated to be approx. 100,000–130,000 from the amino acid composition. A similar value was obtained by determining the protein content per heme a. The cytochrome molecule was composed of three subunits with molecular weights of 55,000, 29,000 and 19,000, respectively. The 29 kd subunit had heme c.Hemes a and c of cytochrome a ₁ c ₁ were reduced on addition of nitrite, and the reduced cytochrome was hardly autoxidizable. Exogenously added horse heart cytochrome c was reduced by nitrite in the presence of cytochrome a ₁ c ₁; K _m values of cytochrome a ₁ c ₁ for nitrite and N. agilis cytochrome c were 0.5 mM and and 6 M, respectively. V _max was 1.7 mol ferricytochrome c reduced/min·mol of cytochrome a ₁ c ₁ The pH optimum of the reaction was about 8. The nitrite-cytochrome c reduction catalyzed by cytochrome a ₁ c ₁ was 61% and 88% inhibited by 44M azide and cyanide, respectively. In the presence of 4.4 mM nitrate, the reaction was 89% inhibited. The nitrite-cytochrome c reduction catalysed by cytochrome a ₁ c ₁ was 2.5-fold stimulated by 4.5 mM manganous chloride. An activating factor which was present in the crude enzyme preparation stimulated the reaction by 2.8-fold, and presence of both the factor and manganous ion activated the reaction by 7-fold.Cytochrome a ₁ c ₁ showed also cytochrome c-nitrate reductase activity. The pH optimum of the reaction was about 6. The nitrate reductase activity was also stimulated by manganous ions and the activating factor.     

Gas Exchange Responses to CO2 Concentration Instantaneously Elevated in Flag Leaves of Winter Wheat Cultivars Released in Different Years

Three winter wheat (Triticum aestivum L.) cultivars, representatives of those widely cultivated in Beijing over the past six decades, were grown in the same environmental conditions. Net photosynthetic rate (P _N) per unit leaf area and instantaneous water use efficiency (WUE) of flag leaves increased with elevated CO₂ concentration. With an increase in CO₂ concentration from 360 to 720 µmol mol^–1, P _N and WUE of Jingdong 8 (released in 1990s and having the highest yield) increased by 173 and 81 %, while those of Nongda 139 (released in 1970s) increased by 88 and 66 %, and Yanda 1817 (released in 1945, with lowest yield) by 76 and 65 %. Jingdong 8 had the highest P _N and WUE values under high CO₂ concentration, but Yanda 1817 showed the lowest P _N. Stomatal conductance (g _s) of Nongda 139 and Yanda 1817 declined with increasing CO₂ concentration, but g _s of Jingdong 8 firstly went down and then up as the CO₂ concentration further increased. Intercellular CO₂ concentration (C _i) of Jingdong 8 and Nongda 139 increased when CO₂ concentration elevated, while that of Yanda 139 increased at the first stage and then declined. Jingdong 8 had the lowest C _i of the three wheat cultivars, and Yanda 1817 had the highest C _i value under lower CO₂ concentrations. However, Jingdong 8 had the highest P _N and lowest C _i at the highest CO₂ concentration which indicates that its photosynthetic potential may be high.     

黄花棘豆脱水素OoY_2K_4的鉴定及表达分析

该研究以黄花棘豆cDNA为模板,采用同源克隆法,从黄花棘豆转录组数据库中克隆获得1个响应逆境胁迫的胚胎发育晚期丰富蛋白基因,命名为OoY_2K_4;OoY_2K_4基因ORF为786bp,编码261个氨基酸,含有2个保守的Y片段和4个K片段,为典型的Y_2K_4类脱水蛋白亚家族成员;OoY_2K_4蛋白不具有跨膜结构域,不存在信号肽,亲水性极强,含有1个糖基化位点和17个磷酸化位点;亚细胞定位显示,OoY_2K_4蛋白定位于细胞质中。多序列比对发现,OoY_2K_4蛋白与其他物种第二组LEA蛋白(脱水素)序列高度保守;进化树分析显示,该序列与三叶草、蒺藜苜蓿和紫花苜蓿相似度最高,亲缘关系最近。采用qRT-PCR对OoY_2K_4基因在干旱、高盐、低温以及脱落酸、乙烯、赤霉素处理下的表达分析显示,干旱和高盐胁迫可显著诱导OoY_2K_4基因表达,而低温胁迫下基本无变化;激素处理均可诱导OoY_2K_4基因高效表达,其中脱落酸诱导下OoY_2K_4基因表达最显著。研究推测,OoY_2K_4基因可能通过依赖ABA的信号途径参与黄花棘豆对干旱和高盐逆境胁迫的应答反应。     

Genes determining leucine aminopeptidase and mildew resistance from the ornamental apple, ‘White Angel’

Mildew resistance in the ornamental apple White Angel was found to be determined by complementary genes. The gene R _w was found to be necessary for the expression of resistance controlled by the resistance gene Pl _w . The close linkage between the isoenzyme gene, Lap-2, for leucine aminopeptidase and P1 _w was confirmed. The efficiency of Lap-2 as a marker in screening for mildew resistance is limited, as it cannot account for susceptible plants with the r _w r _w P1 _w p1 _w genotype. It has, however, an important role to play in combining resistance genes from different sources. The genotypes of White Angel (R _w r _w , Pl _w pl _w , Lap-2an), Jester (R _w r _w , p1 _w p _w , Lap-2an) Katja (R _w r _w ,p1 _w p1 _w , Lap-2an) and Gloster 69 (r _w r _w , p1 _w p1 _w , Lap-2an) were determined. It also appeared that R _w might influence Lap-2 activity in young seedlings.     

Expression of the mosquitocidal cryIVB gene under the control of different promoters in Bacillus sphaericus 2362 and acrystalliferous Bacillus thuringiensis subsp. israelensis c4Q2-72

The 3.7 kb XbaI fragment harbouring the cryIVB gene which encoded a 130 kDa mosquitocidal toxin protein from Bacillus thuringiensis subsp. israelensis (B.t.i.) was placed downstream to the cat-86 gene promoter (P _cat-86, spore stage specific expression) or bgaB gene promoter (P _bgaB , vegetative stage specific expression). The constructs were subcloned into pBC16 to obtain pBTC3 and pBTC6, respectively. Both plasmids and the other construct, pBTC1 were successfully transferred into B. thuringiensis subsp. israelensis c4Q2-72 and B. sphaericus 2362. Western blot analysis showed that P _bgaB in front of P _cryIVB could enable cells to produce a 130 kDa protein from the vegetative stage (4 h) whereas those with P _cat-86 could not. The positive detection of 130 kDa crystal protein during the vegetative stage (4 h) by Western blot analysis indicated the vegetative-stage-specific expression of P _bgaB , while the 130 kDa crystal protein produced from cryIVB gene under control of P _cat-86 was detected only at 48 h. The strong activity of P _bgaB , together with P _cryIVB within pBTC6 in both bacterial hosts was also shown by the toxicity assay against Aedes aegypti larvae (B.t.i. c4Q2-72, 5.6 ± 3.6 × 10² c.f.u./ml; B. sphaericus 2362, 5.4 ± 2.5 × 10² c.f.u./ml) which were 100-fold and 10-fold more toxic to such larvae when compared with pBTC3 (P _cat-86 together with P _cryIVB ) and pBTC1 (contained only its self promoter) in the same bacterial host strains, respectively. The plasmid pBTC6 is not stable in either Bacillus host.     

The chloroplast cytochrome b 6 f complex can exist in monomeric and dimeric states

The cytochrome b ₆ f complex isolated from spinach chloroplast membranes can be resolved into two forms, a monomeric and a dimeric form, by centrifugation on sucrose gradients. The conversion of the dimeric form of the complex into the monomeric form could be prevented by cross-linking with the homobifunctional reagent, dithiobis(succinimidylpropionate) but not by cross-linking with disuccinimidyltartrate or glutaraldehyde. SDS-PAGE analyses of the monomeric and dimeric forms of the cytochrome complex showed the presence of specific cross-linked products in each respective form of the complex. For example, the monomeric form contained a cross-linked product of cytochrome f, cytochrome b ₆ f and subunit IV while the dimeric form contained a cross-linked dimer of cytochrome b ₆ f. The presence of the former in the isolated cytochrome b ₆ f complex prepared by the method of Hurt and Hauska (Eur J Biochem 117: 591–599, 1981) indicates the presence of the monomer in his preparation.Abbreviations DBMIB 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone - DSP dithiobis(succinimidylpropionate) - DST disuccinimidyltartrate