植物中草酸积累与光呼吸惭醇酸代谢的关系

对几种Ｃ３和Ｃ４植物中草酸含量及相应的乙醇酸氧化酶活性测定结果表明,叶片光呼吸强度及其着急酶活性大小与草酸积累量没有相关性;植物根中均能积累草酸,但未测出乙醇酸氧化酶活性。烟草根、叶中的草酸含量在不同生长时期差异明显,且二者呈显著正相关（ｙ＝２．５６５ｌｎｘ＋２．１３７,ｒ＝０．７４９,Ｐ〈０．００１）,说明振中到可能来自叶片。氧化乙醇酸的瓣活性与氧化乙醛酸的酶的活性呈极显著线性正相关（ｙ＝０．２     

荞麦与大豆叶片中草酸含量差异及其可能的原因

用1/5浓度Hoagland营养液培养荞麦和大豆幼苗l0 d后,荞麦叶片、根及其分泌物中的草酸含量均明显高于大豆,说明荞麦叶片的草酸形成能力强.荞麦叶片中存在少量的草酸氧化酶活性,而大豆中未检测到该酶活性,表明荞麦具有一定的降解草酸的能力.乙醇酸氧化酶(GO)催化乙醇酸氧化的活性两种植物之间虽差异不明显,但该酶催化乙醛酸氧化的活性荞麦显著高于大豆.荞麦GO对乙醛酸的Km值明显低于大豆GO,同时其乙醛酸含量也较高,因此其叶片中由乙醛酸形成草酸的速率应高于大豆.由此认为,由乙醛酸氧化生成草酸可能是植物草酸合成限速步骤之一,其反应速率高低可能导致不同种类植物叶片中草酸含量的差异.     

水稻叶片中的草酸积累及其与抗白叶枯病的关系(简报)1

测定不同生长时期及感染白叶枯病菌前后,水稻叶片中的草酸含量、乙醇酸氧化酶活性变化的结果,进一步证实乙醇酸氧化酶同时具有氧化乙醛酸的活性,但叶片中的内源草酸含量变化与乙醇酸氧化酶活性变化无关。高感品种玉梅153和高抗品种中二占在染病前后内源草酸含量变化之间并无显著差异。     

水稻叶片中的草酸积累及其与抗白叶枯病的关系（简报）

测定不同生长时期及感染白叶枯病菌前后,水稻叶片中的草酸含量、乙醇酸氧化酶活性变化的结果,进一步证实乙醇酸氧化酶同时具有氧化乙本钱酸的活性,但叶片另的内源草酸含量变化与乙醇酸氧化酶活性变化无关,高感品种玉梅１５３和高抗品种中二占在染病前后内源草酸含量变化之间并无显著差异。     

荞麦与大豆叶片乙醇酸氧化酶的纯化和性质比较研究

分别从荞麦与大豆叶片中部分纯化了乙醇酸氧化酶 (GO ,EC1 .1 .3 .1 ) ,并研究其部分性质。结果显示荞麦与大豆叶片中GO的催化特性有明显差异 :大豆叶片中GO对乙醇酸Km值为 0 .3 1mmol/L ,对乙醛酸Km值为 1 .98mmol/L。外源草酸对GO氧化乙醇酸活性影响很小 ,但对其氧化乙醛酸活性抑制明显 ,5mmol/L草酸可抑制 44%。而荞麦叶片中GO性质有所不同 :GO对乙醇酸Km为 0 .46mmol/L ,对乙醛酸Km为 0 .85mmol/L。草酸对荞麦GO氧化乙醇酸活性影响也很小 ,对其氧化乙醛酸活性的抑制作用明显小于大豆 ,5mmol/L草酸只抑制 2 4%。上述研究结果表明 ,荞麦GO对乙醛酸的亲和力明显强于大豆 ,并且草酸对其GO氧化乙醛酸活性影响较小。因此相对于大豆而言 ,GO可能在荞麦叶片草酸合成中起重要作用。     

植物叶片中抗坏血酸含量与草酸积累的关系

不同植物和不同生长期烟草叶片中抗坏血酸含量变化与相应的草酸含量变化之间都无显著的相关性;喂饲外源抗坏血酸后的水稻和荞麦叶中草酸含量提高不明显。据此推测：尽管不能排除抗坏血酸可能是植物草酸合成的前体,但其内源含量高低不一定影响植物中草酸积累。     

缺磷胁迫下烟草中草酸含量的变化（简报）

用1／2Hoagland营养液培养6个不同烟草品种,于缺磷后不同时期,测定根、叶中草酸含量的结果显示,在处理后10d时的根和叶中草酸含量均开始上升,并随胁迫时间的延长而增大,其中以中烟90和牛津26提高最为显著。根中未测出乙醇酸氧化酶活化,叶中此酶的活性较高,但不受缺磷胁迫的影响;缺磷处理的根和叶中异柠檬酸裂解酶活性变化也不明显。     

杂交水稻及其亲本光合特性的研究 Ⅲ.功能叶片碳代谢中一些酶活性

对杂交水稻青优159（母本青A,父本R159）和广优四号（母本广A,父本青六矮）及其亲本功能叶片光合碳代谢中一些酶活性进行了研究。实验结果表明了两组杂交水稻的RuBPCase活性、RuBPCase/RuBPOase活性比值超过其各自的亲本;而杂交水稻青化159和广优四号的RuBPOase活性、乙醇酸氧化酶活性、光呼吸速率、光呼吸速率与光合速率的比值明显的低于其各自双亲．对RuBPCase活性和乙醇酸氧化酶活性与光合速率进行相关分析,结果表明RuBPCase活性与光合速率有正相关关系,相关系数为0．768;而乙醇酸氧化酶活性与光合速率负相关,相关系数为-0．834;两者均达到α＝0．05显著水平。     

植物抗逆性与光呼吸作用之间的关系

用Na2SO3溶液浸泡,可以影响菠菜叶片中超氧物歧化酶的活性。较低浓度(0.1、1、10ppm)的该溶液使酶活性增加,而100ppm的该溶液则降低此酶的活性。同时发现叶片中光呼吸关键酶,即乙醇酸氧化酶活性也随着发生正相关的变化。讨论植物受SO32-胁迫后,乙醇酸氧化酶的活性变化与植物抗逆性的产生可能存在密切的关系。     

几种有机酸对芸苔光呼吸代谢的影响（简报）

芸苔叶圆片在乙醇酸氧化酶受抑对照光,其乙醇酸的积累受而酮酸促进,而被柠檬酸、琥珀酸及苹果酸抑制、离体条件下．添加苹果酸、柠檬酸和琥珀酸时,乙醇酸氧化酶活性增高：添加丙酮酸、柠檬酸、苹果酸和琥珀酸时．甘氨酸氧化酶活性也增大。     

乙醇酸氧化酶必需精氨酸残基的化学修饰

丁二酮能使GAO迅速失活,其失活速度受介质pH和硼酸浓度的显著影响;其修饰反应具可逆性,当透析除去修饰剂和硼酸时,活性得到恢复。失活进程表现为假一级动力学。而计算表明,酶的每一活性中心单位与一分子丁二酮结合便可引起酶的失活。底物和竞争性抑制剂均能有效地保护酶免于失活。氨基酸分析表明,酶的失活是因为丁二酮修饰了精氨酸残基。丁二酮修饰GAO后使酶的K_m增大,而V_m没有变化。     

The formation of oxalate from glycolate in rat and human liver

In this study, we attempted to elucidate the metabolic pathway and enzymes actually involved in oxalate formation from glycolate in rat and human liver. In rat liver, the formation of oxalate from glycolate appeared to take place predominantly via glyoxylate. The oxalate formation from glycolate observed with crude enzyme preparations was almost entirely accounted for by the sequential actions of glycolate oxidase and xanthine oxidase (XOD) or lactate dehydrogenase (LDH). Under the conditions used, no significant activity was attributable to glycolate dehydrogenase, an enzyme reported to catalyze the direct oxidation of glycolate to oxalate. Among the three enzymes known to catalyze the oxidation of glyoxylate to oxalate, glycolate oxidase and XOD showed much lower activities (a higher Km and lower Vmax) toward glyoxylate than those with the respective primary substrates. As to LDH, none of the LDH subunit-deficient patients examined showed profoundly lowered urinary oxalate excretion. Based on the results obtained, the presumed efficacies in vivo of individual enzymes, as catalysts of glyoxylate oxidation, and the in vivo conditions assumed to allow their catalysis of oxalate production are discussed.     

光呼吸代谢物乙醇酸、乙醛酸和草酸对烟草叶片硝酸还原的影响

乙醇酸、乙醛酸和草酸能明显促进烟草(Nicotiana rustica)叶片在黑暗中的硝酸还原,光呼吸抑制剂a-羟基吡啶甲烷磺酸能消除前二者的促进作用而不能完全消除草酸的作用。草酸+NAD~+能显著促进离体的硝酸还原。烟叶提取液加入草酸和NAD~+后生成NADH和CO_2认为活体内由乙醛酸氧化生成的草酸是经脱氢生成NADH供硝酸还原之用。未能证明在烟叶内存在乙醇酸脱氨酶,因此排除由乙醇酸直接脱氢以还原硝酸的可能。     

Biogenesis of oxalate in plant tissues

Red beet root discs aerated in potassium phosphate for 2 to 3 days and young spinach leaves actively produce oxalate. A series of labeled compounds was supplied to each of these tissues to determine the extent of conversion to oxalate. Similar results were obtained with the 2 tissues except that in the leaf tissue glyoxylate and glycolate were outstandingly good precursors. Carbon from glucose, acetate, and particularly from some acids of the tricarboxylic acid cycle was recovered in oxalate. Extracts from both tissues were found to contain an enzyme which converts oxaloacetate to oxalate and acetate. The enzyme was partially purified and some of its properties are described. A pathway of oxalate synthesis which does not include glycolate or its oxidase is therefore proposed.     

Synthesis of oxalic Acid by enzymes from lettuce leaves

A rapid purification of lactate dehydrogenase and glycolate oxidase from lettuce (Lactuca sativa) leaves is described. The kinetics of both enzymes are reported in relation to their possible roles in the production of oxalate. Lettuce lactate dehydrogenase behaves like mammalian dehydrogenase, catalyzing the dismutation of glyoxylate to glycolate and oxalate. A model is proposed in which glycolate oxidase in the peroxisomes and lactate dehydrogenase in the cytosol are involved in the production of oxalate. The effect of pH on the balance between oxalate and glycolate produced from glyoxylate suggests that in leaves lactate dehydrogenase may function as part of an oxalate-based biochemical, pH-stat.     

不同因子对荞麦中草酸含量的影响

用不同化合物从根部喂养麦幼苗,测定其根叶中草酸含量的变化。结果表明：异柠檬酸、抗坏血酸及其前体物均可不同程度地降低荞麦根叶中草酸含量;而乙醇酸与乙醛酸则显著提高其草酸含量,表明荞麦叶片草酸合成主要来自乙醇酸途径,而非来自抗坏血酸等途径。水培条件下,以铵态氮或尿素等作唯一氮源时,荞麦中草酸含量远低于以硝态氮培养的;将谷氨酸或丝氨酸加到含硝态氮培养液中也能显著降低其草酸含量,不同氮素影响荞麦草酸含量可能与乙醇酸途径有关。     

Active site and loop 4 movements within human glycolate oxidase: implications for substrate specificity and drug design

Human glycolate oxidase (GO) catalyzes the FMN-dependent oxidation of glycolate to glyoxylate and glyoxylate to oxalate, a key metabolite in kidney stone formation. We report herein the structures of recombinant GO complexed with sulfate, glyoxylate, and an inhibitor, 4-carboxy-5-dodecylsulfanyl-1,2,3-triazole (CDST), determined by X-ray crystallography. In contrast to most alpha-hydroxy acid oxidases including spinach glycolate oxidase, a loop region, known as loop 4, is completely visible when the GO active site contains a small ligand. The lack of electron density for this loop in the GO-CDST complex, which mimics a large substrate, suggests that a disordered to ordered transition may occur with the binding of substrates. The conformational flexibility of Trp110 appears to be responsible for enabling GO to react with alpha-hydroxy acids of various chain lengths. Moreover, the movement of Trp110 disrupts a hydrogen-bonding network between Trp110, Leu191, Tyr134, and Tyr208. This loss of interactions is the first indication that active site movements are directly linked to changes in the conformation of loop 4. The kinetic parameters for the oxidation of glycolate, glyoxylate, and 2-hydroxy octanoate indicate that the oxidation of glycolate to glyoxylate is the primary reaction catalyzed by GO, while the oxidation of glyoxylate to oxalate is most likely not relevant under normal conditions. However, drugs that exploit the unique structural features of GO may ultimately prove to be useful for decreasing glycolate and glyoxylate levels in primary hyperoxaluria type 1 patients who have the inability to convert peroxisomal glyoxylate to glycine.     

Oxalate accumulation from citrate by Aspergillus niger. I. Biosynthesis of oxalate from its ultimate precursor.

Carbon-14 was incorporated from citrate-1,5-14C, glyoxylate-14C(U), or glyoxylate-1-14C into oxalate by cultures of Aspergillus niger pregrown on a medium with glucose as the sole source of carbon. Glyoxylate-14C(U) was superior to glyoxylate-1-14C and citrate-1,5-14C as a source of incorporation. By addition of a great amount of citrate the accumulation of oxalate was accelerated and its maximum yield increased. In a cell-free extract from mycelium forming oxalate from citrate the enzyme oxaloacetate hydrolase (EC3.7.1.1) was identified. Its in vitro activity per flask exceeded the rate of in vivo accumulation of oxalate. Glyoxylate oxidizing enzymes (glycolate oxidase, EC1.1.3.1; glyoxylate oxidase, EC1.2.3.5;NAD(P)-dependent glyoxylate dehydrogenase; glyoxylate dehydrogenase, CoA-oxalylating, EC1.2.1.7) could not be detected in cell-free extracts. It is concluded that in cultures accumulating oxalate from citrate after pregrowth on glucose, oxalate arises by hydrolytic cleavage of oxaloacetate but not by oxidation of glyoxylate.