氨基酸转运载体研究进展

氨基酸转运载体是介导氨基酸跨膜转运的膜蛋白,在氨基酸营养机体细胞和神经调节过程中起着重要作用;而且,其功能异常会导致严重的氨基酸吸收和代谢障碍性疾病,也具有重要的病理学意义。本文就近年来关于中性氨基酸、酸性氨基酸和碱性氨基酸转运载体家族成员及其组织分布、分子生物学特征、生理功能和病理学意义等研究进展进行了综述。     

酚酸类物质对三七幼苗的化感影响

该文研究了不同浓度的阿魏酸、对香豆酸、丁香酸、对羟基苯甲酸、香草酸5种酚酸类物质对三七幼苗生长和生理的影响。结果表明:处理后,三七幼苗的苗高、根长、可溶性蛋白质含量、根系活力、CAT以及POD活性均有所降低。其中,阿魏酸各处理组幼苗的苗高及POD活性均显著降低,50、100 mg·L~(-1)的对香豆酸以及100 mg·L~(-1)的香草酸处理组幼苗苗高也分别比对照显著降低16.19%、16.67%和29.29%;对香豆酸、丁香酸以及对羟基苯甲酸各处理组幼苗根长均显著低于对照;香草酸处理组幼苗的根系活力也显著低于对照,且幼苗的CAT活性在10、50、100 mg·L~(-1)丁香酸、对羟基苯甲酸以及香草酸处理下也达到了显著降低水平。此外,1 mg·L~(-1)阿魏酸以及100 mg·L~(-1)香草酸处理组幼苗的叶绿素含量也均显著降低;中高浓度的阿魏酸、对香豆酸、丁香酸、对羟基苯甲酸增加了三七幼苗的MDA含量,而香草酸在0.1、1、10、100 mg·L~(-1)浓度下显著降低幼苗的MDA含量;丁香酸、香草酸、对羟基苯甲酸以及中高浓度的对香豆酸增加了三七幼苗的SOD活性,且香草酸各处理组均达到了显著性水平。综上结果表明,5种酚酸类物质对三七幼苗均具有一定的化感抑制作用,但各酚酸物质的作用方式及强度并不完全一致,阿魏酸的化感影响较大,这为进一步研究三七的化感自毒作用提供了一定的理论参考。     

海藻酸盐裂解酶研究进展

海藻酸盐裂解酶是一类降解褐藻中海藻酸盐的酶。此酶已经在多种有机体中得到分离。对海藻酸盐裂解酶的生物特性、研究方法及其生物学功能进行了介绍。在酶学特性研究的基础上 ,通过酶解构建新型海藻酸盐多聚物 ,可增强和扩展海藻酸盐裂解酶在工业、农业、医药领域中的应用 ,使其在海藻多糖的高值化应用中发挥重要的作用。概述了海藻酸盐和海藻酸盐裂解酶过去和现在的研究状况 ,展望了海藻酸盐和海藻酸盐裂解酶将来的应用前景。     

Biotechnological routes based on lactic acid production from biomass

Lactic acid, the most important hydroxycarboxylic acid, is now commercially produced by the fermentation of sugars present in biomass. In addition to its use in the synthesis of biodegradable polymers, lactic acid can be regarded as a feedstock for the green chemistry of the future. Different potentially useful chemicals such as pyruvic acid, acrylic acid, 1,2-propanediol, and lactate ester can be produced from lactic acid via chemical and biotechnological routes. Here, we reviewed the current status of the production of potentially valuable chemicals from lactic acid via biotechnological routes. Although some of the reactions described in this review article are still not applicable at current stage, due to their “greener” properties, biotechnological processes for the production of lactic acid derivatives might replace the chemical routes in the future.     

D-焦谷氨酸的研制

研究了Ｄ－焦谷氨酸的制备,提纯精制以及分析化验等方面的有关问题。     

Sucrose phosphorylases catalyze transglycosylation reactions on carboxylic acid compounds

Two sucrose phosphorylases were employed for glycosylation of carboxylic acid compounds. Streptococcus mutans sucrose phosphorylase showed remarkable transglycosylating activity, especially under acidic conditions. Leuconostoc mesenteroides sucrose phosphorylase exhibited very weak transglycosylating activity. Three main products were detected from the reaction mixture using benzoic acid and sucrose as an acceptor and a donor molecule, respectively. These compounds were identified as 1-O-benzoyl α-d-glucopyranoside, 2-O-benzoyl α-d-glucopyranose, and 2-O-benzoyl β-d-glucopyranose by 1D-and 2D-NMR analyses of the isolated products and their acetylated products. Time-course analyses proved that 1-O-benzoyl α-d-glucopyranoside was initially produced by the transglycosylation reaction of the enzyme. 2-O-Benzoyl α-d-glucopyranose and 2-O-benzoyl β-d-glucopyranose were produced from 1-O-benzoyl α-d-glucopyranoside by intramolecular acyl migration reaction. S. mutans sucrose phosphorylase showed broad acceptor-specificity. This sucrose phosphorylase catalyzed transglycosylation to various carboxylic compounds such as short-chain fatty acids, hydroxy acids, dicarboxylic acids, and phenolic carboxylic acids. 1-O-Acetyl α-d-glucopyranoside was also enzymatically synthesized by transglucosylation reaction of the enzyme. The sensory test of acetic acid and the glucosides revealed that the sour taste of acetic acid glucosides was significantly lower than that of acetic acid.     

Lipid composition of the pea aphid,Acyrthosiphon pisum (Harris) (homoptera: Aphididae), reared on host plant and on artificial media

Polar and neutral lipids and their constitutive fatty acids were quantified in the pea aphid, Acyrthosiphon pisum (Harris), grown on host plant or on a lipid free artificial diet. The results were compared to determine if lipids were involved in the suitability of the diet for continuous rearing of this A. pisum biotype. For apterous adults grown on plants, the lipids were characterized by a low amount of neutral lipids (2.5% weight/fresh weight) almost entirely (96.4%) composed of hexanoyl and sorboyl dimyristin. These storage lipids were higher in the alatae (3.8%), probably correlated with potential flight activity. The phospholipid amounts were identical in these two morphs (1.3–1.4% weight/fresh weight), comprised mainly of phosphatidylethanolamines (52%) and phosphatidylcholines (40.6%). These phospholipids contained a still unidentified fatty acid, with a retention time close to that of linolenic acid and synthesized by the aphid or its bacterial symbionts (not found in plants). The apterous adult aphids reared on an artificial diet showed an accumulation of neutral lipids (8.9% for the first generation); this increase was shown to be slightly greater for the hexanoyl and sorboyl triglycerides. In contrast, the phospholipids decreased in aphids reared on an artificial diet (1.1% and 0.9%, respectively, for first and second generation), correlated with a phospholipid fatty acid profile significantly deficient in C18:3 and in the unidentified peculiar fatty acid. These phospholipids are essential components of biological membranes and a diet-driven deficient synthesis in some of their components may result in the observed symptoms. © 1992 Wiley-Liss, Inc.     

尼泊尔杞木的化学成分研究

从尼泊尔杞木(Alus nepalensis)地上部分的95%乙醇提取物中首次分离到9个化合物,应用波谱方法或与已知品对照的手段鉴定它们为乙酰齐墩果酸(1)、齐墩果酸(2)、羽扇豆醇(3)、β-谷甾醇(4)、β-香树酯醇(5)、euseaphic acid(6)、喜树碱(7)β-胡萝卜甙(8)、绿原酸(9).尼泊尔杞木是喜树碱第十一个植物来源.喜树碱在尼泊尔杞木的干燥叶和茎中的含量分别是923和192μg/g.     

牛磺熊去氧胆酸的合成进展

牛磺熊去氧胆酸是由熊去氧胆酸的羧基和牛磺酸的氨基之间缩水而形成的结合型胆汁酸。主要介绍了目前四种牛磺熊去氧胆酸合成方法,并对四种方法的优缺点进行了全面的评价,从而为牛磺熊去氧胆酸的合理开发和利用提供思路。     

氨基酸L-和D-异构体对离体小麦胚植株生长的影响

5种氨基酸L-和D-异构体对离体小麦胚植株生长的影响结果表明,在1~7 mmol.L-1浓度范围内,脯氨酸异构体均不抑制植株生长;缬氨酸和蛋氨酸的L-型严重抑制生长,D-型不抑制或轻微抑制;丙氨酸则相反,L-型无抑制作用,D-型是严重抑制类型;丝氨酸的L-型轻微抑制生长,D-型严重抑制生长。生长结果也表明,DL-异构体的抑制作用介于D-型和L-型之间。     

生物催化富马酸加氨合成天门冬氨酸的研究进展

天门冬氨酸是构成蛋白质的基本氨基酸之一,也是一种重要的医药工业原料。天门冬氨酸酶催化富马酸加氨合成天门冬氨酸。本文从天门冬氨酸酶的结构与性质、产酶菌的筛选与培养条件优化、酶促反应动力学研究、酶的分子生物学研究4个方面介绍了国内外关于生物转化合成天门冬氨酸的研究概况。     

Auxin effects on rooting in pea cuttings

Light-grown stem cuttingss of Pisum sativum L. cv. Weibull's Marma were rooted in a nutrient solution. The presence of 10 μ M indolylacetic acid (IAA) in the solution for 24 h or longer periods decreased the number of roots subsequently formed to about 50% of control, provided IAA was present in the solution during any of the 4 first 24 h periods. Treatment for 6 h or shorter periods caused no or small response. IAA did not appreciably change the time needed for root formation, the time course of root appearance or the pattern of root distribution along the basal internode. IAA at 100 μ M usually increased the number of roots although variable results were obtained with this IAA concentration.
The number of roots was strongly increased by treatment with indolylbutyric acid (IBA) or 2,4-dichlorophenoxyacetic acid (2,4-D). None of these or other synthetic auxins decreased the number of roots in suboptimal concentrations. Experiments with 10 μ M IBA showed that stimulation of rooting was obtained only if the auxin was present in the rooting solution for several days. Simultaneous treatment with IAA decreased the stimulating effect of IBA to some extent, whereas no such response was obtained if IAA was combined with 2,4-D.
IAA applied in lanolin to the stem of intact cuttings decreased the number of roots formed. Decapitation and debudding of the cuttings decreased the number of roots formed. If at least 2 leaves were left this decrease was efficiently counteracted by an optimal IAA dose applied to the upper part of the stem. A five times higher dose was less effective, indicating a negative effect on rooting also by IAA applied to the shoots.     

Effects of Rat Ovariectomy on CSF Monoamine Metabolite Levels and Elimination

Cerebrospinal fluid (CSF) was removed from the third ventricle of anesthetized male, female, and ovariectomized rats. CSF 3,4-dihydroxyphenylethylamine and serotonin metabolite levels [dihydroxyphenylacetic acid, homovanillic acid (HVA), and 5-hydroxyindoleacetic acid (5-HIAA)] were determined on 15-min samples by liquid chromatography coupled with electrochemical detection. Monoamine oxidase inhibition was used for studying metabolite turnover in the CSF. No difference was observed between male, ovariectomized, and sham-operated female rats. However, ventricular CSF HVA and 5-HIAA levels were significantly higher in the ovariectomized than in the sham-operated rats. These differences do not reflect effects of ovariectomy on brain metabolite production but indicate slower metabolite elimination from the CSF.     

RNA—RNA原位杂交实验条件探讨

将Vigilin和qroal(Ⅰ)cDNA亚克隆到pGEM3Z和pGEM4Z载体,体外转录合成35s标记的cRNA探针。经RNA凝胶电泳,Southern Northern,杂交检查探针长度,杂交特性和特异性,通过系列实验探讨了RNA-RNA原位杂交实验中固定、杂交前处理、杂交温度,探针量、探针长度,洗脱严格性和RNA酶处理等对杂交结果的影响,建立了简化的RNA-RNA原位杂交方法。     

Studies on Biological Activity of Cyclic Imide Compounds

The structure-activity relationships of l-phenylpyrrolidine-2,5-dione derivatives were investigated on Sclerotinia sclerotiorum by the agar dilution method. In addition, several representative compounds were tested for antimicrobial spectra in vitro with 15 pathogenic microbes and for foliage protection activity in green house tests with rice blast, rice brown spot, rice sheath blight and kidney bean stem rot. It was found that 3,5-dihalo-substituents on the benzene moiety are essential to high antifungal activity against Sclerotinia sclerotiorum. Generally, l-(3′,5′-dihalophenyl)pyrrolidine-2,5-diones are active against Corticiaceae, Dematiaceae, Pleosporaceae and Sclerotiniaceae, especially active against Sclerotinia sclerotiorum and Botrytis cinerea (the conidia form of Sclerotinia fuckeliana). N-(3,5-Dichlorophenyl)itaconimide showed a peculiarly broad antimicrobial spectrum. In green house tests, these compounds showed high activity against rice brown spot, rice sheath blight and kidney bean stem rot. Results of green house tests on the above-mentioned diseases correlate fairly well with those of in vitro tests.     

Effect of phenolic acids on growth of Chlorella pyrenoidosa

Static bioassays were conducted on Chlorella pyrenoidosa using vanillic acid, syringic acid, and 4-hydroxybenzoic acid. At 0.1 mM, vanillic and 4-hydroxybenzoic acid stimulated cultural growth compared to the control. For both compounds, concentrations of 0.3 mM and 0.4 mM were initially inhibitory, then after 3–5 days became stimulatory compared to control. Bioassays with syringic acid at 0.1 mM, 0.3 mM, and 0.4 mM were all stimulatory. At 0.5 mM–2.5 mM, syringic acid resulted in 100% mortality in C. pyrenoidosa. Bacteria-free cultures of C. pyrenoidosa were stimulated by vanillic acid at 0.1 mM and inhibited at 0.4 mM with no shift in response observed. It was suggested that degradation of test material is responsible for the shift from inhibition to stimulation. All three compounds are reported to change from enzyme synergists to antagonists as concentrations are reduced. Because these compounds are major components of humic material, it is suggested that they have the potential to confound studies involving interaction of toxins and humics.     

Short-term fatty acid effects on adipocyte glucose uptake: Mechanistic insights

The modulation of insulin sensitivity in visceral fat tissue could be important in the treatment of Type 2 diabetes mellitus. Selected fatty acids may impact on insulin-stimulated and basal glucose uptake in adipocytes, thus isolated rat epididymal adipocytes were exposed to 100 μM oleic, arachidonic, eicosapentaenoic, docosahexaenoic or stearic acids and insulin (15 nM) or vehicle for 30 min. Glucose uptake was quantified by measuring uptake of ³H-deoxyglucose/mg adipocyte protein/min. Where appropriate, inhibitors were included to elucidate the mechanisms involved.In this model, insulin stimulated glucose uptake with 62±7%. All fatty acids tested, except for stearic acid, depressed insulin-stimulated glucose uptake by an average of 33±4.2%. On the other hand, all fatty acids tested except stearic and arachidonic acids, stimulated basal glucose uptake with an average of 34±8.1%. Inhibitor studies showed the involvement of prostaglandins, lipoxins, protein kinase C and tyrosine kinase in these processes.     

Effects of abscisic acid on CAM in Portulacaria afra

Water stress induces Crassulacean acid metabolism (CAM) in Portulacaria afra as manifested by day stomatal closure, organic acid fluctuation, and night CO₂ uptake. We now have evidence that abscisic acid treatment of leaves causes partial stomatal closure that is accompanied by the induction of CAM in a manner similar to water stress. There appears to be an inverse relationship between exogenous CO₂ uptake and decarboxylation of organic acids in that organic acids remain high during the day providing stomata are open. When stomata close, there is consumption of organic acids by decarboxylation. The hypothesis is that stomatal opening controls CAM in this species.This material is based upon work supported by the Science and Education Administration of the USDA under Competitive Grant No. 5901-0410-8-0018-0.     

Maternal grazing on stubble and Mediterranean shrubland improves meat lipid profile in light lambs fed on concentrates

Concentrates-fed lamb meat is often associated with an unfavourable lipid profile (high levels of saturated and/or n-6 polyunsaturated fatty acids; SFA and PUFA). For this reason, Spanish sheep producers from Mediterranean areas are turning to traditional grazing by ewes to obtain healthier lamb meat. The objective of this research was to determine the effects of maternal grazing on the fatty acid (FA) composition of weaned lamb meat. The ewes (Segureña breed) were allocated to two different rearing systems during pregnancy (5 months) and lactation (45 days): (i) feeding indoors on barley grain and lucerne pellets; (ii) grazing on cereal stubble, fallow land and seasonal pastures consisting of Mediterranean shrubs, herbs and trees. Two groups of 20 autumn and spring lambs were sampled. The lambs were weaned at 13.1±0.9 kg and 45.0±4.1 days age and fed on grain-based concentrates until they reached 24.8±2.1 kg live weight (light lambs slaughtered at 98.3±3.6 days of age). The FA content was determined in the intramuscular loin fat by gas chromatography using a flame ionization detector. The ewe diet did not affect the levels of the main lamb FAs (C18:1c+t, C16:0 and C18:2c), and so did not provide any additional reduction in fat saturation. Saturated fatty acids represented around 40% of total FAs determined in the meat. Ewe grazing acted as an n-3 PUFA-promoting diet, providing a lamb meat with a lower n-6/n-3 ratio. Spring lamb meat had higher proportions of n-3 PUFA (C18:3n-3, C20:5, C22:5 and C22:6) and conjugated linoleic acid (C18:2c9t11+c11t9) to the detriment of the n-6 PUFAs (C20:4, C20:2 and C22:4), while autumn lamb meat also had higher levels of C18:3n-3 and C18:3n-6, and lower level of C20:4, which points to little seasonal differences. The n-6/n-3 ratio achieved by ewe grazing fell from 8.2 to 4.1 (Spring) and from 7.6 to 5.5 (Autumn), values which are close to those recommended in human diet for good cardiovascular health. These n-6/n-3 reductions were associated with lower levels of total PUFA and C20:4n-6. Our research concluded that grazing on stubble and Mediterranean shrubland by ewes, a sustainable rearing practice involving local agro resources, contributed to obtaining weaned lamb meat with a more favourable lipid profile and so can be recommended to sheep farmers.