水分胁迫对露花磷酸烯醇式丙酮酸羧化酶表达水平及特性的影响

水分胁迫能引进露花叶片ＰＥＰ羧化酶的活力,酶蛋白和ｍＲＮＡ水平的提高。复水后,叶片ＰＥＰ羧化酶表达量降低,茎中的ＰＥＰ羧化酶在水分胁迫和恢复水分供应过程中变化情况与叶片相似,兼性ＣＡＭ植物的碳代谢类型转变发生在植物的绿色组织中。     

几种CAM植物PEP羧化酶同工酶的比较研究

比较研究几种兼性和专一性ＣＡＭ植物材料的ＰＥＰＣ同工酶表明：经自然干旱诱导,兼性ＣＡＭ植物露花（Ｍｅｓｅｍｂｒｙａｎｔｈｅｍｕｍｃｏｒｄｉｆｏｌｉｕｍ）、长药景天（Ｓｕｄｕｍｓｐｅｃｔａｂｉｌｅ）有新的ＰＥＰＣ同工酶的出现,诱导前后各同工酶的天然分子量变化不大;而土三七（Ｓｅｄｕｍａｉｚｏｏｎ）则没有新的ＰＥＰＣ同工酶出现,但诱导后其同工酶的天然分子量有所增大。以上几种兼性ＣＡＭ植物的ＰＥＰＣ同工酶酶谱无明显昼夜变化。专一性ＣＡＭ植物的ＰＥＰＣ酶谱和天然分子量均较一致,亦无昼夜差异。     

大豆C4途径与光系统Ⅱ光化学功能的相互关系

测定了不同发育时期大豆（Ｇｌｙｃｉｎｅ ｍａｘ（Ｌ）Ｍｅｒｒ．）“黑农４１”叶片的４种Ｃ４酶（ＰＥＰＣａｓｅ（磷酸烯醇式丙酮酸羧化酶）、ＮＡＤＰ－ＭＤＨ（ＮＡＤＰ苹果酸脱氢酶）、ＮＡＤＰ－ＭＥ（ＮＡＤＰ苹果酸酶）和ＰＰＤＫ（丙酮酸磷酸二激酶））活性、荧光动力学数值（Ｆｖ／Ｆｏ（ＰＳⅡ活性）、ｑＰ（光化学淬灭）、ｑＮ（非光化学淬灭、ΦＰＳⅡ（有效ＰＳⅡ光化学效率））和光合速率。结果表明在“黑农４１”     

曲霉N1—14‘胞质酶活性与产L—苹果酸能力的关系

Ｌ－苹果酸（ＬＭＡ）高产突变株曲霉Ｎ１－１４’在高产酸状态下,其胞质中催化ＣＯ２固定反应的酶有四种：丙酮酸羧化酶（ＰＣ）、磷酸烯醇丙酮羧化酶（ＰＥＰＣ）、磷酸烯醇丙酮酸羧化激酶（ＰＣＫ）和苹果酸酶（ＭＥ）;除ＭＥ之外,三种羧化酶的活性与ＬＭＡ产生速率呈较好的线性正相关关系;苹果酸脱氢酶（ＭＤＨ）活性比ＰＣ等酶高２￣３个数量级;琥珀酸脱氢酶（ＳＤＨ）活力则明显低,几种酶只有ＳＤＨ与发酵醪中ＬＭＡ含量     

C-末端酰胺化酶的研究进展

Ｃ－末端酰胺化酶的研究进展王翔林（沈阳药学院沈阳１１００１５）Ｃ－末端酰胺化酶（Ｃａｒｂｏｘｙｌ－ｔｅｒｍｉｎａｌα－ＡｍｉｄａｔｉｎｇＥｎｚｙｍｅ,ｃ－ＡＥ）,又称甘氨肽酰化单氧酶（Ｐｅｐｔｉｄｙｌｇｌｙｃｉｎｅα－Ａｍｉｄａｔｉｎｇｍｏｎｏｏｘｙｇｅｎａｓｅ,ＰＡＭ,ＥＣ１．１４．１７．１３）是动物体内普遍存在的一种酶。     

菠萝叶片PEP羧激酶活性及草酰乙酸与腺苷酸库的昼夜变化

自然条件下生长的菠萝植株,其叶片的ＰＥＰ羧激酶的脱羧与可逆羧化活性于中午１２时达最大值。脱羧／羧化比值在上午明显高于下午和晚上。脱羧反应底物ＯＡＡ含量于夜间增高,至次日早晨６时最高,随后下降。白天的ＯＡＡ／Ｍａｌ值高于夜间。参与Ｍａｌ及ＯＡＡ形成转化的ＰＥＰＣ和ＭＤＨ活性的昼夜变化与ＰＥＰ羧激酶（ＰＥＰＣＫ）活性的图式相似。ＰＥＰ羧激酶脱羧反应的另一底物ＡＴＰ在夜间０～６时内处于低水平,白天９～１５时内则显著增高。     

暖温带地区几种木本植物碳稳定同位素的特点

通过对暖温带落叶阔叶林生态系统主要乔、灌木植物叶片、树干、花、果实碳稳定性同位素比率（δ＾１３Ｃ值）测定和比较,发现这些植物叶片的δ＾１３Ｃ值受多种因子的影响,具有较大的种间差异及时空异质性,主要表现在不同植物δ＾１３Ｃ值不同;种内δ＾１３值变化也很大,以荆条的差异最大,为６．５４９‰（－２２．２２６‰－－２８．７７５‰）;其次是大叶白蜡５．７０６‰（－２３．６８７‰－－２９．３９３‰）;核桃秋５     

野生经济植物资源橡籽仁可利用价值的研究

以京白种鸡作鸡作试验动,以玉米、稻谷作对照,采用鸡真代谢能（ＴＭＥ）法研究野生经济植物资源橡胶籽仁的可利用营养价值。结果表明,橡籽仁１ｋｇ含量总能（ＧＥ）１６．５３ＭＪ,表观代谢能（ＡＭＥ）１１．１３ＭＪ,真代谢能（ＴＭＥ）１１．６６ＭＪ,,含粗蛋白（ＣＰ）１０．６３％,ＣＰ的表观利用率（ＣＰＡＡ）４５．５５％,真利用率（ＣＰＴＡ）４９．８３％,１７种氨基酸（ＡＡ）总含量９．２３％,必需氨基酸（Ｅ     

CaM BP－10对NAD激酶的抑制效应

ＮＡＤ激酶在光合作用等植物生理过程中起重要作用。ＮＡＤ激酶的激活依赖于钙离子和钙调素（ＣａｌｍＯｄｕｌｉｎ,ＣａＭ）．从植物中分离得到的一种新的ＣａＭ结合蛋白ＣａＭＢＰ－１０（ＢＰ－１０）明显抑制ＮＡＤ激酶的激活活性,抑制作用可被ＣａＭ所克服．动力学研究表明,抑制效应是ＢＰ－１０与ＣａＭ之间特异性相互作用的结果。实验证实ＢＰ－１０对ＮＡＤ激酶活性起着重要调节作用．     

干旱胁迫对红松幼苗保护酶活性及脂质过氧化作用的影响

随着土壤的逐渐干旱,红松（Ｐｉｎｕｓ ｋｏｒａｉｅｎｓｉｓ Ｓｉｅｂ．ｅｔ．Ｚｕｃｃ）幼苗叶中膜质过氧化产物丙二醛（ＭＤＡ）含量和膜相对透性均在干旱处理后第３天迅速上升;组织自动氧化速率先是增加,在干旱处理第３天后恢复到处理前水平。保护酶ＳＯＤ、ＰＯＤ、ＣＡＴ的活性明显提高,只有ＡＳＰ的活性下降。用ＰＥＧ模拟干旱胁迫与土壤自然干旱胁迫结果略有不同,－１．０ＭＰａ ＰＥＧ溶液对红松幼苗具有较为明显的     

Stable Carbon Isotope Ratio and Activities of PEP Carboxylase and PEP Carboxykinase in Pineapple Leaves

Slight flutuation in carbon isotope values were found in counted from top dounward to the 35th in pineapple, Ananas comosus (L.) Merr., but more negative δ13C value (less heavier 13C) was observed in lower position leaves. The average δ 13C value was –12.94‰ in 11 leaves with maximum range of variation as –2.06‰. Similar single peak curves were found between PEPCase and PEP carboxykinase activities with leaves at various positions. Both enzymes reached the maximum activity in 8—11th leaves, then declined in others at lower positions. PEP carboxykinase activity was 3.4 folds higher than PEPCase activity under the present experimental condition (25—30 ℃). The results indicated that metabolic coordination evisted between dark carboxylation and light decarboxylation. For the obligate CAM plant, pineapple, though the carboxylation and decarxylation activities did occur in old leaves, the CAM level change did much, however.     

植物磷酸烯醇式丙酮酸羧化酶的可逆胍变性

纯化的高梁叶片磷酸烯醇式丙酮酸羧化酶(PEP羧化酶)经不同浓度的盐酸胍处理变性失活后,在试验的蛋白浓度范围内,它的失活时间进程的动力学分析表明为一级反应。0.4 M盐酸胍处理25分钟后(O℃),酶的催化活性完全丧失,酶蛋白的远紫外圆二色性光谱、内源荧光光谱及免疫特异性等测定均表明酶的结构发生了深刻变化。甘油及PEP羧化酶的变构效应剂G6P和甘氨酸对酶在盐酸胍溶液中的变性作用有一定的保护效果。变性酶用复性缓冲液稀释20倍后,在最佳条件下,再经30分钟保温,酶的催化活性能恢复70％以上。G6P和甘氨酸能促进变性酶的复性,甘油亦有明显效果。随着酶活性的恢复,它的远紫外圆二色性、内源荧光及免疫特异性也随之恢复,变性酶的复性速率在常温下(25℃)比在低温下(0℃)要快得多。     

Expression, purification, and characterization of a bacterial GTP-dependent PEP carboxykinase

The Corynebacterium glutamicum (C. glutamicum) phosphoenolpyruvate carboxykinase (PCK) gene (pckA) was cloned into an Escherichia coli expression vector with a glutathione S-transferase (GST) tag. This recombinant DNA can produce highly overexpressed tagged protein in soluble form. This is the first report of the production of C. glutamicum PCK overexpressed in E. coli. The GST-fused PCK was purified using the glutathione-Sepharose 4B affinity column and the GST tag was removed in one-step. This one-step, easy purification method would be very useful for future mutational and structural studies. The molecular mass of the purified protein is approximately 68 kDa as confirmed by mass spectrometry and it is a monomeric enzyme. Also, the enzyme assays revealed that C. glutamicum PCK has a GTP-specific activity and that its activity is maximal in the presence of both Mn2+ and Mg2+.     

Relationship between pseudo-HPr and the PEP: fructose phosphotransferase system in Salmonella typhimurium and Escherichia coli

Summary We have studied in Salmonella typhimurium and Escherichia coli the properties of pseudo-HPr suppressor mutations. These mutations suppressed the defects in a ptsH mutant which lacks HPr, one of the enzymes of the phosphoenolpyruvate: carbohydrate phosphotransferase system. The suppressor mutation was mapped in S. typhimurium at 3 min, closely linked to leu. The corresponding chromosomal fragment of 1.7 kb from S. typhimurium and E. coli (extending clockwise from ilvH) was cloned. In a maxicell system a protein with an approximate molecular weight of 36,000 was synthesized. Pseudo-HPr suppressor mutations (fruR) and a deletion extending clockwise from leu resulted in the constitutive expression of the fru operon containing the genes for II^Fru (fruA), III^Fru (fruB), fructose 1-phosphate kinase (fruK) and pseudo-HPr (fruF). fruR probably codes for a repressor of the fru operon. Tn10 mutagenesis revealed the following order of genes in the fru operon: fruB-(fruK, fruF)-fruA. Pseudo-HPr activity could replace HPr in PEP-dependent phosphorylation of PTS carbohydrates. III^Fru could be phosphorylated both via HPr and pseudo-HPr, since mutants lacking pseudo-HPr activity were still able to phosphorylate fructose in the presence of added HPr. Both the pseudo-HPr suppressor mutations at 3 min and the deletion extending from leu had an additional phenotype. Introduction of these mutations or deletions was always accompanied by disappearance of PEP synthase activity. Complementation of such a mutant with the cloned fragments reversed both phenotypes at the same time. Possibly, the fruR gene product acts as an activator of the gene coding for PEP synthase.     

Metabolic networks to generate pyruvate,PEP and ATP from glycerol in Pseudomonas fluorescens

Glycerol is a major by-product of the biodiesel industry. In this study we report on the metabolic networks involved in its transformation into pyruvate, phosphoenolpyruvate (PEP) and ATP. When the nutritionally-versatile Pseudomonas fluorescens was exposed to hydrogen peroxide (H₂O₂) in a mineral medium with glycerol as the sole carbon source, the microbe reconfigured its metabolism to generate adenosine triphosphate (ATP) primarily via substrate-level phosphorylation (SLP). This alternative ATP-producing stratagem resulted in the synthesis of copious amounts of PEP and pyruvate. The production of these metabolites was mediated via the enhanced activities of such enzymes as pyruvate carboxylase (PC) and phosphoenolpyruvate carboxylase (PEPC). The high energy PEP was subsequently converted into ATP with the aid of pyruvate phosphate dikinase (PPDK), phosphoenolpyruvate synthase (PEPS) and pyruvate kinase (PK) with the concomitant formation of pyruvate. The participation of the phospho-transfer enzymes like adenylate kinase (AK) and acetate kinase (ACK) ensured the efficiency of this O₂-independent energy-generating machinery. The increased activity of glycerol dehydrogenase (GDH) in the stressed bacteria provided the necessary precursors to fuel this process. This H₂O₂-induced anaerobic life-style fortuitously evokes metabolic networks to an effective pathway that can be harnessed into the synthesis of ATP, PEP and pyruvate. The bioconversion of glycerol to pyruvate will offer interesting economic benefit.     

PEP car☐ykinase containing species in the Brachiaria group of the subfamily panicoideae

In the Gramineae, a survey of species among the Brachiaria group of the subfamily Panicoideae, tribe Paniceae revealed that they are PEP car?ykinase containing species. This group includes the genera Brachiaria, Eriochloa and Urochloa. With the exception of the genus Panicum, these are the only genera within the Panicoideae found to contain PEP car?ykinase species. It is suggested that the PEP car?ykinase species of the genus Panicum, P. fasciculatum, P. maximum, P. molle and P. texanum, might be best placed in the Brachiaria group.     

甘蓝型油菜PEP基因片段的克隆和种子特异性反义表达载体的构建及转基因油菜的获得

丙酮酸羧化酶(PEP)是控制油菜蛋白质/油脂含量比例的一个种子的含油量.本研究利用PCR法从甘蓝型油菜花油5号(H045)克隆了PEP基因片段,并与载体pBI121-B构建了反义PEP基因的种子特异性植物表达载体,通过激光微束穿刺法将其转化到甘蓝型油菜中,目前已获得了转基因植株.     

Leupaxin binds to PEST domain tyrosine phosphatase PEP

PEST domain tyrosine phosphatase (PEP) is an intracellular protein tyrosine phosphatase and characterized by PEST motifs and proline-rich domains in the carboxyl terminal half. PEP is primarily expressed in hematopoietic cells, and together with PEP-binding Csk, may act as a negative regulator of antigen receptor signaling in lymphocytes. Here, we show the binding capability of PEP for leupaxin, which is preferentially expressed in hematopoietic cells and a comparatively new member of the paxillin family characterized by two protein-protein interaction modules, LIM domains and LD motifs. These results suggested that leupaxin might participate in the regulation of the signaling cascade through the binding to PEP in lymphocytes. (Mol Cell Biochem 269: 13–17, 2005)