乙醇法木质素羟甲基化改性研究

以刺五加根茎剩余物为原料,以乙醇作为提取木质素溶剂,通过单因素实验和正交实验优化乙醇法木质素羟甲基化改性,其最优工艺条件是:碱量2.0%,反应温度80℃,反应时间3.5 h,碱木质素与甲醛质量比3∶1。在此条件下获得羟甲基化改性乙醇法木质素羟甲基含量最高可达到11.56%,游离甲醛含量为0.257%,本研究为实现乙醇法木质素的高附加值资源化应用提供理论和实验依据。     

氮杂环卡宾二氧化碳加合物催化转酯反应制备生物柴油

为了开发一种无金属有机催化剂用于生物柴油的制备,合成了一系列咪唑(啉)类氮杂环卡宾的二氧化碳加合物(N-heterocyclic carbenes CO2adducts,NHC-CO2),通过加热使其释放游离卡宾,并催化转酯反应制备生物柴油。为了比较催化活性,不同结构的NHC—CO2被用于大豆油的转酯反应中。结果发现:当使用咪唑类催化剂时,产物中甲酯含量大于90%,而当使用咪唑啉型催化剂,甲酯含量不足20%,这说明咪唑类催化剂更适合本研究中的转酯反应。催化剂最佳用量为大豆油的2%(摩尔百分比),最佳醇油比为12∶1。本研究中催化剂前体释放游离卡宾进入反应介质,反应迅速,产品分离简单,是制备生物柴油的有效绿色方法。     

制革醛鞣固体废弃物的酶法资源化利用

为建立毛皮制革产生固体废弃物的清洁化、资源化利用工艺,采用JW酶制剂对兔毛皮甲醛鞣制固废进行酶解处理,并对酶解产物的工农业用途进行评估。结果表明,固废中的皮块能够被JW-4酶制剂完全水解。皮块水解后回收的毛纤维经检测其长度优于刀剪工艺,断裂强度、伸长率与原始纤维相当。酶解液中含大量小分子的多肽和氨基酸,氨基酸浓度为5.23%,游离甲醛含量为37.34 mg/kg,重金属含量低于氨基酸肥料标准;将酶解液培养小麦种子后,能够促进小麦提前1 d生根,培养6 d的植株鲜重要优于对照。用清洁产品处理制革固废,不仅避免二次污染,还可对固废进行资源化利用。     

不同的甲醛杀菌浓度对A群、C群脑膜炎球菌荚膜多糖内毒素含量的影响

目的研究不同的甲醛杀菌浓度对A群、C群脑膜炎球菌荚膜多糖内毒素含量的影响。方法将A群脑膜炎球菌发酵液分成A、B两组,A组采用体积分数为2.5%甲醛杀菌,B组采用体积分数为2.0%甲醛杀菌。将C群脑膜炎球菌发酵液分成C、D两组,C组采用体积分数为2.5%甲醛杀菌,D组采用体积分数为2.0%甲醛杀菌。分别纯化获得荚膜多糖,用动态浊度法测定荚膜多糖中内毒素的含量。结果 A、C两组荚膜多糖中内毒素含量显著低于B、D两组荚膜多糖中的内毒素含量(P<0.05)。结论使用不同浓度的甲醛杀菌,对A群、C群脑膜炎球菌荚膜多糖内毒素的含量有显著影响。较高浓度的甲醛利于菌体细胞的固定,从而防止细菌自溶释放内毒素,因而高浓度的甲醛能够减少其内毒素的含量,提高了产品质量。     

脱共生作用对黑豆蚜氨基酸代谢的影响(英文)

为了解共生菌对黑豆蚜蛋白质、氨基酸代谢的影响 ,用利福平处理黑豆蚜以除去其细胞内共生细菌 ,产生脱共生蚜虫。结果表明 ,被脱去共生菌的蚜虫与未经抗生素处理的正常蚜虫相比 ,7日龄时 ,脱共生蚜虫每毫克鲜重的总蛋白含量降低了 2 9% ,每毫克鲜重的游离氨基酸含量提高了 17%。对黑豆蚜取食的蚕豆苗韧皮部组织中必需氨基酸所占的比例进行分析后发现 ,蚕豆苗韧皮部组织中的必需氨基酸含量仅占 2 0 % ,而有共生菌的黑豆蚜组织中必需氨基酸已达到 4 4% ,脱共生后降低到 37% ,这些结果证明了黑豆蚜的胞内共生菌为其寄主提供了部分必需氨基酸。通过对游离氨基酸组成的分析发现 ,在测定的 17种氨基酸中 ,必需氨基酸中的苏氨酸在共生蚜虫中所占的比例为 2 1 6 % ,在脱共生蚜虫中仅为 16 7%。同样 ,非必需氨基酸中的酪氨酸和丝氨酸 ,在共生蚜虫中分别占总游离氨基酸的 8 9%和 5 6 % ,而在脱共生蚜虫中却分别升高到 2 1 1%和 13 6 %。这些结果表明 ,各种氨基酸比例的失调 ,造成了脱共生蚜虫蛋白质合成受阻和部分游离氨基酸的积累 ,并因此导致蚜虫发育和繁殖的失调。     

环己亚胺对盐诱导游离脯氨酸累积的影响

环己亚胺(CHX)单独作用会增加高梁苗中游离脯氨酸的含量,原因可能有:一是CHX抑制了根的正常吸收功能,导致植株失水,游离脯氨酸增加;二是CHX抑制了蛋白质合成,使总的游离氨基酸累积,从而也表现出游离脯氨酸含量的增加,后者可能更为主要。为此,用CHX研究与脯氨酸合成有关的基因活性化或表达时,一定要考虑CHX单独的作用。NaCl诱导的游离脯氨酸的累积可被CHX处理所抑制。在NaCl处理2～4h内加CHX后,抑制效果几乎可达到100％,以后随CHX处理的时间越长,其抑制作用越小。     

兴安落叶松游离氨基酸的动态变化与损伤因子的关系

为研究不同损伤因子及其不同损伤程度对落叶松游离氨基酸组分及其含量的影响,用剪叶和落叶松毛虫(Dendrolimussuperans)取食损伤兴安落叶松(Larixgmelinii)幼苗,使用HPLC柱前AccQ.Tag衍生法测定针叶内游离氨基酸的组成及含量的变化。结果表明,松针中含有17种游离氨基酸,以苯丙氨酸含量最高,损伤15d后,剪叶4枝比对照增加36.6%,虫害4枝增加的百分率达到98.3%。剪叶4枝15d后及虫害4枝5d后,损伤25%与50%之间氨基酸总含量差异显著(P0.05),且昆虫取食4枝25%、50%、75%后松针内游离氨基酸总含量的变化趋势与剪叶处理后总氨基酸含量的变化一致,均在处理10d后呈现较高的含量,虽虫害后的氨基酸总含量高于剪叶损伤后的总含量,但二者之间差异不显著。剪叶4枝、8枝、12枝3个损伤程度后针叶内的游离氨基酸含量无显著差异,说明剪叶4枝即能超过落叶松的耐受阈值,达到防御诱导效应。两种处理下多种游离氨基酸的含量均有下降,其中8种氨基酸含量存在显著差异(P0.05),即丝氨酸、谷氨酸、甘氨酸、组氨酸、脯氨酸、半胱氨酸、赖氨酸和苯丙氨酸。表明人工模拟剪叶的损伤方式与虫害都可诱导松树氨基酸组分及含量的变化,而间接影响害虫的生长发育。     

不同施肥水平对不同品种小麦籽粒蛋白质和地上器官游离氨基酸含量的影响

研究了小麦不同品种开花后各地上器官游离氨基酸含量的变化动态及其与籽粒蛋白质含量的关系,以及氮素营养的调节作用。结果表明,小麦叶片和颖壳十穗轴游离氨基酸含量均在开花后持续增加,至花后14d达到最大值,之后开始下降。茎和叶鞘游离氨基酸含量则在开花后上升较缓慢,至花后21d达最大值。籽粒游离氨基酸含量一般在开花后就持续降低。各地上器官游离氨基酸含量与成熟期籽粒蛋白质含量均呈显著或极显著正相关,说明源器宫中游离氨基酸供应充足,有利于籽粒蛋白质积累。增施氮肥能够提高各地上器官中的游离氨基酸含量,进而促进籽粒蛋白质的合成,提高籽粒蛋白质含量。品种之间籽粒蛋白质含量的差异,是由各地上营养器官向籽粒运输氨基酸的综合作用所造成的。     

酶法催化甲醛合成木酮糖

木酮糖是生物体内的代谢中间产物,是多种稀有糖合成的前体物质,因其独特的生物活性在膳食、保健、医药等领域发挥着重要作用。本研究旨在从最基本有机原料之一的甲醛出发,利用生物酶法催化甲醛合成木酮糖。通过来源于恶臭假单胞菌Pseudomonas putida的苯甲酸脱羧酶(Benzoylformate decarboxylase)突变体BFD-M3催化甲醛聚合生成羟基乙醛和1,3-二羟基丙酮(DHA)。通过来源于大肠杆菌的转醛醇酶(Transaldolase)突变体Tal B-F178Y进一步催化羟基乙醛和DHA聚合生成木酮糖,最终实现甲醛到木酮糖的酶法转化,转化率为0.4%。此外,经过优化甲醛底物浓度,木酮糖转化率达到4.6%,比优化前提高了11.5倍。为了进一步提高木酮糖的转化率,采用Scaffold多酶组装技术固定BFD-M3、Tal B-F178Y蛋白,使木酮糖转化率达到14.02%,较未用Scaffold技术前提高3倍,为生物法合成稀有糖提供了一种新方案。     

过量无机盐及谷氨酸抑制重组菌核黄素发酵

在重组枯草芽孢杆菌24/pMX45核黄素发酵中,酵母粉促进核黄素合成,酵母抽提物抑制核黄素合成。分析显示,酵母抽提物的无机离子和游离氨基酸含量均高于酵母粉。在酵母粉基础发酵培养基中,添加各种无机离子和游离氨基酸,使其含量与酵母抽提物相同。摇瓶发酵结果表明:过量的无机离子和谷氨酸对核黄素合成有显著的抑制作用。酵母抽提物含有较高浓度的谷氨酸,是其抑制核黄素合成的主要原因。     

樟芝发酵液和菌丝体提取物抗氧化性能研究

采用甲醇、乙醇、异丙醇对樟芝发酵液和菌丝体进行提取,对其不同提取物进行还原力,DPPH、羟基自由基、超氧自由基、过氧化氢清除能力,亚铁离子螯合能力等抗氧化能力的测定。比较发酵液提取物和菌丝体提取物的抗氧化效果,比较不同提取剂提取物的抗氧化效果。结果显示:(1)发酵液提取物还原力最高的为异丙醇提取物,亚铁离子螯合能力最强的为乙醇提取物;DPPH清除能力乙醇和异丙醇提取物最高都达到97.88%;羟基自由基清除能力最高的为乙醇提取物,达到84.22%;3种提取物的超氧自由基清除能力都较高,具有最高清除率的异丙醇提取物达到94.89%;过氧化氢清除能力最高的为乙醇提取物的97.53%。(2)菌丝体提取物还原力最高和亚铁离子螯合能力最强的均为乙醇提取物;DPPH清除能力最高的为乙醇提取物的89.95%;3种提取物的羟基自由基清除能力普遍偏低,最高的仅为异丙醇提取物的37.28%;超氧自由基清除能力则普遍较高,最高为甲醇提取物的90.05%;过氧化氢清除能力最高的为乙醇和异丙醇提取物,最高清除率均为95.06%。抗氧化性能的研究比较,为进一步开发利用樟芝提供了依据。     

V-A菌根真菌对柑桔吸收钙素效应的研究初报*

应用放射性同位素~(45)Ca示踪来探明泡囊—丛枝菌根真菌对柑桔吸收钙素的作用。两次盆栽试验,施用含~(45)Ca(NO_3)_2的Hoagland营养液,分别在红壤及无土基质人工接种菌根真菌Glomus citricolum于枳(Poncirus trifoliata)和构头橙(Citrus aurantium)实生苗根系,以不接种为对照。结果表明在红壤中接种2个月后,枳已形成菌根,5个月的菌根感染率为24.5％。在无土基质中,菌根的形成更好;接种3个月后,构头橙的菌根感染率为49.5％,并且发生丰富的泡囊。在红壤中,有菌根枳实生苗的地上部从肥料中吸收的钙素和全钙含量都比无菌根的增加75％左右。在无土基质中,有菌根的枳和构头橙实生苗地上部含钙量及放射性强度都比无菌根的显著增高。放射性自显影直接地显示了菌根真菌将吸收的~(45)Ga运转至地上茎叶中,对构头橙的效应更明显。     

93份贵州地方辣椒资源品质性状分析

对93份贵州地方辣椒资源的品质性状进行了分析,结果表明:粗纤维含量在22.99%~44.06%之间,牛角椒含量最高,为33.71%;粗脂肪含量在11.49%~27.46%之间,锥形椒含量最高,为21.55%;蛋白质含量在12.81%~22.97%之间,指形椒含量最高,为18.78%;辣椒素含量在0.28~7.61 mg/g之间,指形椒含量最高,为4.47 mg/g;4个指标的平均隶属度值在0.21~0.68之间,超过0.60的辣椒资源有S106、S103、S072和S016;聚类分析表明,93份辣椒资源按平均隶属度大小可分为四大类型,即高品质型、中品质型、一般品质型和特殊品质型。贵州地方辣椒种质资源品质性状分析可为辣椒种质资源的创新利用、新品种的选育奠定基础。     

化妆品防腐剂布罗波尔的稳定性及其释放甲醛的研究

研究发现化妆品防腐剂布罗波尔在水中会因pH增加、储存温度增高和放置时间的增长等因素,在水中缓慢分解且释放出少量甲醛。在含布罗波尔的化妆品中,采用乙酰丙酮比色法测得甲醛含量,比采用高效液相色谱法测得甲醛要高。此外,还发现28 d内的布罗波尔可释放甲醛的含量比较低,引起化妆品中甲醛超标的风险性较小。     

AN ARTEFACT IN RADIOAUTOGRAPHY DUE TO BINDING OF FREE AMINO ACIDS TO TISSUES BY FIXATIVES

The binding of labeled free amino acids to liver and to purified protein by commonly used fixatives was investigated. Glutaraldehyde caused 25% of free leucine to be bound to serum albumin in solution, whereas formaldehyde bound only 0.5%. Liver slices were incubated for 2 min in the presence of labeled leucine and of puromycin, which permits absorption of leucine into the cell but inhibits incorporation into protein. Both counting and radioautographic techniques showed that glutaraldehyde bound 30 times, and osmic acid six times, as much free amino acid as did formaldehyde. By comparing liver slices incubated with and without puromycin for 2 min, it was calculated that in radioautographs prepared after fixation with glutaraldehyde, osmic acid, or formaldehyde 63, 25, and 4% respectively of the grains were due to binding of free amino acid. Formaldehyde, freshly prepared from paraformaldehyde, gives good preservation and is the recommended fixative for radioautography. When levels of free substrate in a tissue are high at the time fixative is added, the amount of binding of free substrate induced by the fixative should be included as a control in radioautographic experiments.     

两阶段氮源补料策略促进微拟球藻EPA高效积累

为促进微拟球藻EPA高效积累,本研究探索了光照强度和氮源种类对微拟球藻生理生化及EPA相对含量的影响;根据光照和氮源实验结果设计两阶段氮源补料策略,并优化了氮源补料时间。结果表明:光强从1 000 lux增加至9 000 lux,微拟球藻比生长速率从0.25 d~(-1)增加至0.54 d-1,油脂含量从11.4%增加至20.2%,而EPA相对含量从25.5%降低至13.1%;培养10 d后乙酸铵组生物量和油脂含量最高,分别为0.41 g·L~(-1)和14.3%,硝酸钠组EPA相对含量最高,为27.1%;两阶段硝酸钠加乙酸铵培养模式,最佳乙酸铵补料时间为第11 d,培养20 d最终生物量及EPA相对含量分别为0.68 g·L~(-1)和24.3%。采用两阶段氮源补料策略能促进微拟球藻高效积累EPA。     

Formaldehyde metabolism in semicarbazide intoxication

Subchronic administration of semicarbazide in the experiment with the rats was used to reduce the formaldehyde level in the organism in order to reveal the interaction between formaldehyde metabolism and biochemical parameters, which define the oxidant-antioxidant system condition and NO metabolism. It has been found that under semicarbazide impact the generation of free radicals, ROS, nitrite and nitrate were enhanced while aldehydes level was reduced that resulted from not only semicarbazide effect like the aldehydes acceptor, but the formaldehyde synthesis slowdown and acceleration of its transformation into format as well. We suppose that formaldehyde plays certain role in the development of connective tissue pathology.     

珊瑚菜不同部位香豆素含量的研究

应用高效液相色谱技术,对珊瑚菜（Glehnia littoralis Fr.Schmidt ex Miq.）的果实、叶和根以及采后去皮入药的根和根皮中的补骨脂素、欧前胡素和异欧前胡素3种香豆素的含量进行了测定。结果显示：（1）3种香豆素在果实、叶与根、根皮中均有积累,总含量在果实中最高,为0.6364mg·g-1,根中为0.0657mg·g-1,根皮中为0.0312mg·g-1,叶中为0.0151mg·g-1,采后去皮处理的根中最低,仅为0.0081mg·g-1。（2）珊瑚菜根经水烫去皮处理后香豆素含量急剧下降,与同期采收未去皮处理的根相比,去皮后根内香豆素总含量、补骨脂素、欧前胡素、异欧前胡素含量分别下降了87.7%、100%、82.76%、85.25%。（3）与未去皮的根相比,处理后的根皮中香豆素总量、补骨脂素、欧前胡素和异欧前胡素含量分别是未去皮处理的珊瑚菜根中的47.42%、31.37%、51.54%和53.28%。研究表明：从充分利用香豆素成分的角度出发,根入药时应带根皮使用,另外,叶和根皮不应丢弃,均可收集作为提取香豆素新的植物资源,果实中香豆素含量很高,亦可作为香豆素资源。     

Some aspects of the Feulgen reaction in situ

Summary Cytological observations combined with studies on absorption spectra of Feulgen stained normal and lipid — extractet HeLa and ehrlich-Lettré mouse ascites cells were performed after fixation of the cells as well in neutral formaldehyde as in Serra fixative. The effects of formaldehyde treatment of the stained cells to substitute all the free amino groups of DNA bond pararosaniline molecules, were also studied. The results obtained by using DNA samples containing 2% protein and relatively free from protein, led to the conclusion that after acid hydrolysis for a short period purines in DNA become splitted and these released aldehydes react with one or two amino groups of pararosaniline, a triphenylmethane dye (according to the arrangement of purines and pyrimidines in the helices). Some protein molecules also take part in the reaction and substitute some of the free amino groups of DNA bound pararosaniline. Peulgen stained cells fixed in Serra fixative show an absorption maximum at 546–550 m. Under appropriate conditions, as in cells fixed in formaldehyde, other substances e.g. phospholipids and lipoproteins interfere with the reaction by substituting most of the free amino groups of DNA bound pararosaniline molecules. It has been argued that in histochemical reactions monosubstituted pararosaniline molecules should be coloured and further substitution of free amino groups of pararosaniline, bound in DNA helices, does not change the intensity of the colour, but gives a shift in the wavelength of the absorption spectra.It has been suggested that the differential response of the nucleoli to the Feulgen-reaction, depending on whether the cells were fixed in formaldehyde or in Serra fixative, may be due to the formation of a protecting shield around the finely distributed intranucleolar chromatin strands, when formaldehyde is being used. After this fixation lipoproteins and other lipids, present in a relatively high percentage and closely associated with the intranucleolar chromatin strands, are especially well preserved.Evidences have been put foreward in support of the amino alkylsulfonic acid theory of Rumpf (1935) and Hörmann et al. (1958) whereas the amino sulfinic acid theory to explain the Schiffs reaction (Wieland and Scheuing, 1921) was shown not to be in agreement with our results.On leave from the Department of Botany, Calcutta University, 35, Ballygunge Circular Road, Calcutta-19, India; on a fellowship from the German Academic Exchange Service.     

A free radical mechanism of prostaglandin synthase-dependent aminopyrine demethylation

The mechanism of prostaglandin synthase-dependent N-dealkylation has been investigated using an enzyme preparation derived from ram seminal vesicles. Incubation of an N-alkyl substrate, aminopyrine, with enzyme and arachidonic acid, 15-hydroperoxyarachidonic acid, or tert-butyl hydroperoxide resulted in the formation of the transient aminopyrine free radical species. Formation of this radical species, which was detected by electron paramagnetic resonance spectroscopy and/or absorbance at 580 nm, was maximal approximately 30 s following initiation of the reaction and declined thereafter. Free radical formation corresponded closely with formaldehyde formation in this system, in terms of dependence upon substrate and cofactor concentration, as well as in terms of time course. Both aminopyrine free radical and formaldehyde formation were inhibited by indomethacin and flufenamic acid, inhibitors of prostaglandin synthase. The results suggest that the aminopyrine free radical is an intermediate in the prostaglandin synthase-dependent aminopyrine N-demethylase pathway. The aminopyrine free radical electron paramagnetic resonance spectrum revealed that this species is a one-electron oxidized cation radical of the parent compound. A reaction mechanism has been proposed in which aminopyrine undergoes two sequential one-electron oxidations to an iminium cation, which is then hydrolyzed to the demethylated amine and formaldehyde. Accordingly, the oxygen atom of the aldehyde product is derived from neither molecular nor hydroperoxide oxygen, but from water.