Responses of soil extracellular enzyme activities to carbon input alteration and warming in a subtropical evergreen broad-leaved forest

土壤胞外酶来源于土壤微生物、植物和动物, 是土壤生物地球化学过程的积极参与者, 在森林生态系统的物质循环和能量流动过程中扮演着重要角色。为探明土壤胞外酶活性对碳输入变化及增温的响应, 该研究基于长期增温、去除地表凋落物(以下简称去凋)和切根处理的云南哀牢山亚热带常绿阔叶林控制实验平台, 研究了不同处理(对照、去凋、切根、切根并增温)下表层矿质土壤(0-5和5-10 cm)与碳氮磷获取相关的胞外酶活性, 包括多酚氧化酶(POX)、过氧化物酶(PER)、β-葡萄糖苷酶(BG)、β-1,4-N-乙酰氨基葡萄糖苷酶(NAG)和酸性磷酸酶(AP)。结合铵态氮(NH₄⁺-N)含量、硝态氮(NO₃^--N)含量、溶解有机碳(DOC)含量、溶解总氮(DN)含量、土壤含水量(SWC)等相关指标, 探讨凋落物碳输入、根系碳输入和温度变化对土壤胞外酶活性及其生态化学计量特征的影响。研究结果表明: 在对照样方, 除POX外其余酶活性均为0-5 cm层显著高于5-10 cm层。与对照相比, 长期的凋落物去除显著降低了0-5 cm层土壤AP和BG活性, 对NAG、PER和POX活性则无显著影响; 长期切根处理显著降低了0-5 cm层土壤BG活性, 但提高了两个土层PER活性; 长期切根并增温处理显著降低了0-5 cm层AP和BG活性, 对其余酶活性无显著影响。冗余分析结果显示SWC和NH₄⁺-N含量是驱动土壤酶活性变化的重要因子。本研究为该生态系统土壤碳氮磷生物地球化学关键过程对全球变化的响应提供了土壤酶学的依据。     

亚热带常绿阔叶林土壤胞外酶活性对碳输入变化及增温的响应

土壤胞外酶来源于土壤微生物、植物和动物, 是土壤生物地球化学过程的积极参与者, 在森林生态系统的物质循环和能量流动过程中扮演着重要角色。为探明土壤胞外酶活性对碳输入变化及增温的响应, 该研究基于长期增温、去除地表凋落物(以下简称去凋)和切根处理的云南哀牢山亚热带常绿阔叶林控制实验平台, 研究了不同处理(对照、去凋、切根、切根并增温)下表层矿质土壤(0-5和5-10 cm)与碳氮磷获取相关的胞外酶活性, 包括多酚氧化酶(POX)、过氧化物酶(PER)、β-葡萄糖苷酶(BG)、β-1,4-N-乙酰氨基葡萄糖苷酶(NAG)和酸性磷酸酶(AP)。结合铵态氮(NH₄⁺-N)含量、硝态氮(NO₃^--N)含量、溶解有机碳(DOC)含量、溶解总氮(DN)含量、土壤含水量(SWC)等相关指标, 探讨凋落物碳输入、根系碳输入和温度变化对土壤胞外酶活性及其生态化学计量特征的影响。研究结果表明: 在对照样方, 除POX外其余酶活性均为0-5 cm层显著高于5-10 cm层。与对照相比, 长期的凋落物去除显著降低了0-5 cm层土壤AP和BG活性, 对NAG、PER和POX活性则无显著影响; 长期切根处理显著降低了0-5 cm层土壤BG活性, 但提高了两个土层PER活性; 长期切根并增温处理显著降低了0-5 cm层AP和BG活性, 对其余酶活性无显著影响。冗余分析结果显示SWC和NH₄⁺-N含量是驱动土壤酶活性变化的重要因子。本研究为该生态系统土壤碳氮磷生物地球化学关键过程对全球变化的响应提供了土壤酶学的依据。     

Comparison of substrate specificities of Escherichia coli endonuclease III and its mouse homologue (mNTH1) using defined oligonucleotide substrates

Escherichia coli endonuclease III (Endo III) and its eukaryotic homologues are major repair enzymes for pyrimidine lesions formed by reactive oxygen species and ionizing radiation. In the present study, the activities of Endo III and its mouse homologue (mNTH1) have been compared using defined oligonucleotide substrates containing a urea residue (UR), two cis-thymine glycol (TG) diastereoisomers, 5, 6-dihydrothymine (DHT), and 5-hydroxyuracil (HOU). The substrates were incubated with Endo III and mNTH1, and their activities were compared based on the product analysis by gel electrophoresis. Endo III recognized all base lesions tested, but the activity for DHT was extremely lower than other substrates. In contrast, albeit some preference of UR, mNTH1 showed essentially comparable activities for all substrates including DHT. Comparison of the enzymatic parameters for cis-TG and DHT revealed that large decreases in the affinity (K(m), 27-fold) and k(cat) (11-fold) relative to cis-TG made DHT an very poor substrate for Endo III. mNTH1 had comparable affinities and k(cat) for both cis-TG and DHT, though turnover (k(cat)) of mNTH1 was notably slower than Endo III. In view of the reaction mechanism, the paired base effect on the damage recognition by the two enzymes was also examined. The activities of Endo III for UR and HOU were paired base-independent, but those for cis-TG and DHT were significantly enhanced when paired with G. With mNTH1, the paired base effect was evident only for DHT. The variations of the repair activity with paired bases and enzymes are discussed in relation to the base flipping mechanism suggested for base excision repair enzymes.     

The development of gluconeogenic enzymes in the liver and kidney of fetal and newborn foals.

The activities of glucose-6-phosphatase (G6P), fructose diphosphatase, phosphoenolpyruvate carboxykinase (PEPCK), aspartate and alanine transferases were measured in liver and kidney of fetal foals between 100-318 days of gestation (term approximately 335 days) and during the immediate postnatal period (0-48 h after birth). All 5 enzymes could be detected in the fetal liver and kidney at the youngest gestational age studied. Mean fetal activities were lower than those observed in their mothers and showed no change with gestational age for the majority of enzymes studied. However, renal PEPCK and renal and hepatic G6P did increase towards term. At birth, hepatic and renal activities of these two enzymes were higher than those found in late gestation or in the adult animals. There was no apparent change in the activities of any of the other enzymes at birth. In late gestation (80-90% gestation), the activities of G6P and PEPCK in the foal were low compared to those in other species at the same stage of gestation. Similarly, the perinatal increase in enzyme activity occurred closer to term in the foal than in most other species. These observations indicate that maturation of glucogenic capacity occurs relatively late in the fetal foal and suggests that this process may be dependent on the prepartum rise in fetal cortisol as occurs in other species.     

Acetyl-CoA Synthesizing Enzymes in Cholinergic Nerve Terminals

The activities of five enzymes involved in acetyl-CoA synthesis, pyruvate dehydrogenase complex, ATP citrate lyase, carnitine acetyltransferase, acetyl-CoA synthetase, and citrate synthase, were determined in normal nucleus interpeduncularis and nucleus interpeduncularis in which cholinergic terminals were removed following lesion of the habenulointerpeduncular tract. The activities of aspartate transaminase, fumarase, and GABA transaminase also were determined to compare the effect of lesion on other mitochondrial enzymes which are not linked to the biosynthesis of ACh. In normal nucleus interpeduncularis the activities of carnitine acetyltransferase and pyruvate dehydrogenase complex were higher than the activity of ChAT (choline acetyltransferase), whereas the activities of acetyl-CoA synthetase and citrate synthase were considerably lower than that of ChAT. The effect of the lesion separated the enzymes into two groups: the activities of pyruvate dehydrogenase complex, carnitine acetyltransferase, fumarase and aspartate transaminase decreased by 30--40%, whereas the activities of the other enzymes descreased 5--15%. ChAT activity was in all cases less than 15% of normal. It could be concluded that none of the acetyl-CoA synthesizing enzymes decreased to the degree that ChAT did. Only pyruvate dehydrogenase complex and carnitine acetyltransferase seem to be localized in cholinergic terminals to a significant degree. ATP citrate lyase as well as acetyl-CoA synthetase seem to have less significance in supporting acetyl-CoA formation in cholinergic nerve terminals.     

Effect of carbon and nitrogen growth limitation upon nutrient uptake and metabolism in batch cultures of Catharanthus roseus (L) G. Don

Cell suspension cultures of the Madagascan Periwinkle, Catharanthus roseus (L). G. Don were grown as batch cultures in two different types of media; in one medium the limiting nutrient was inorganic nitrogen, and in the other it was carbon. The response of the cells to these growth-limiting conditions was monitored by measuring cellular fresh weight, dry weight and protein accumulation, cell viability, medium sugar and nitrate levels, and the activities of certain intracellular enzymes throughout growth in batch culture. The enzymes investigated were glucose-6-phosphate dehydrogenase (EC 1.1.1.49), 6-phosphogluconate dehydrogenase (EC 1.1.1.44), hexokinase (EC 2.7.1.40), phosphofructokinase (EC 2.7.1.11), nitrate reductase (EC 1.6.6.1), glutamate dehydrogenase (EC 1.4.1.2) and glutamine synthetase (EC 6.3.1.2). The effect of culturing the cells under different nutritional regimes was apparent in all aspects of growth; only some enzyme activities were unaffected. Cell viability remained at a high level for several days after growth limitation in both types of culture. The possibility that protein degradation in nitrogen-limited batch cultures is under very stringent control is discussed.     

Comparative study of the antioxidant defence systems in the erythrocytes of australian marsupials and monotremes

A comparison of the erythrocyte (RBC) antioxidant metabolites and enzymes in nine marsupial and two monotreme species was carried out. Reduced glutathione (GSH) concentrations were comparable with those reported for other marsupial and eutherian species. An important finding was that the erythrocytes of the southern hairy nosed wombat regenerated GSH faster than the erythrocytes from its close relative, the common wombat. The activities of glutathione-S-transferase, NADH-methaemoglobin reductase, Superoxide dismutase, and glutathione peroxidase (GSH-Px), showed similar levels and extents of variation as those observed in other marsupial and eutherian species. Catalase activities in the marsupials were lower than those measured in the two monotreme species and much lower than those reported in eutherian species. A negative correlation, significant at P < 0.05, was observed between GSH-Px and catalase activities in the RBC of the marsupials. Since both these enzymes “detoxify” H₂O₂, there appears to be a reciprocal relationship between the activities of these enzymes in marsupial RBC     

Long-term feeding effect of thermally oxidised oils on antioxidant enzymes in rats

Long term feeding effect of thermally oxidised oils on activities of hepatic antioxidant enzymes viz. catalase, glutathione peroxidase (GPX), glutathione-S-transferase (GST) and superoxide dismutase (SOD) at 5 and 20% level for 20 weeks was studied in laboratory rats. Activity of catalase increased in heated and fried oil fed group of rats, whereas activities of GPX, GST and SOD decreased in both heated and fried oil groups. Increase or decrease in activities of these enzymes may be related to several factors like heating and frying conditions, nature of fat, extent of peroxidation, presence of antioxidants, duration of feeding, beside other factors.     

Some properties of acid and alkaline phosphates from boar sperm plasma membranes

Boar sperm plasma membranes were purified by differential and sucrose density equilibrium centrifugation and were found to yield a single band at a density of 1.14 g/cm3. Both alkaline and acid phosphatase activities were enriched in this fraction. The alkaline phosphatase activity was optimal in 100 mM tris (hydroxymethyl) methylamine (Tris)-NaHCO3 at pH 9.9 with 0.05% Triton X-100 and 1 mM MgCl2. This activity was inhibited by ethylenediaminetetraacetic acid (EDTA), cadmium, zinc or heating at 60 degrees C for 30 min. Also, L-homoarginine caused approximately 70% inhibition and L-phenylalanine or L-leucine caused about 10 to 20% inhibition. Acid phosphatase activity was optimal in 100 mM sodium acetate at pH 5.1 with 0.05% Triton. Sodium dodecyl sulfate, potassium fluoride (KF) or sulfhydryl reagents inhibited the activity, while EDTA or heating at 60 degrees C had no effect. These data for enzymes from boar sperm plasma membranes can be used for future work on the quantitation of the enzymes, distinguishing these two phosphatases from other phosphohydrolases, purification of the enzymes and for comparison to phosphatases in other tissues.     

Changes in digestive enzymes through developmental and molt stages in the spiny lobster, Panulirus argus

Changes in major digestive enzymes through developmental and molt stages were studied for the spiny lobster Panulirus argus. There were significant positive relationships between specific activity of trypsin and amylase enzymes and lobster size, whereas esterase and lipase specific activities decreased as lobsters aged. No relationship was found between amylase/trypsin ratio and lobster size. Positive trends were found, however, for trypsin/lipase and amylase/lipase ratios. Results suggest that changes in enzyme activity respond to the lobsters' physiological needs for particular dietary components although multivariate analysis suggested that enzyme activities could be not totally independent of diet. On the other hand, the pattern of changes of major enzyme activities through molt cycle was similar for most enzymes studied. Following molt, trypsin, chymotrypsin, amylase, and lipase activities gradually increased to maximal levels at late intermolt (C4) and premolt (D). There were no variations in the electrophoretic pattern of digestive enzymes through developmental and molt stages and thus, it is demonstrated that regulation is exerted quantitatively rather than qualitatively. Further studies on the effect of other intrinsic and extrinsic factors on digestive enzyme activities are needed to fully understand digestive abilities and regulation mechanisms in spiny lobsters.     

Peptidohydrolases of Soybean Root Nodules : IDENTIFICATION, SEPARATION, AND PARTIAL CHARACTERIZATION OF ENZYMES FROM BACTEROID-FREE EXTRACTS

Nodule extracts prepared from Glycine max var Woodworth possessed endopeptidase, aminopeptidase, and carboxypeptidase activities. Three distinct endopeptidase activities could be resolved by disc-gel electrophoresis at pH 8.8. According to their order of increasing electrophoretic mobility, the first of these enzymes hydrolyzed azocasein and n-benzoyl-l-Leu-beta-naphthylamide, while the second hydrolyzed n-benzoyl-l-Arg-beta-naphthylamine (Bz-l-Arg-betaNA), n-benzoyl-l-Arg-p-nitroanilide (Bz-l-Arg-pNA), and azocasein. The third endopeptidase hydrolyzed Bz-l-Arg-betaNA, Bz-l-Arg-pNA, and hemoglobin. Fractions of these enzymes extracted from electrophoresis gels were shown to have pH optima from 7.5 to 9.8. All of the endopeptidases were completely inhibited by diisopropylphosphorofluoridate, demonstrating that they were serine proteases.Aminopeptidase activity was measured using amino acyl-beta-naphthylamides. Electrophoresis of nodule extracts at pH 6.8 resolved the aminopeptidase activity of nodule extracts into at least four fractions based on mobility and on activities toward amino acyl-beta-naphthylamides. The major activity of two of the aminopeptidases was directed toward l-Leu- and l-Met-beta-naphthylamide, while the other two aminopeptidases exhibited broader specificity and were capable of hydrolyzing a large number of amino acyl-beta-naphthylamides. Two of the aminopeptidases extracted from electrophoresis gels were classified as thiol type enzymes, and all four aminopeptidases had neutral to basic pH optima.     

Regulation of enzyme synthesis in the arginine biosynthetic pathway of Pseudomonas aeruginosa.

In Pseudomonas aeruginosa the synthesis of only two out of eight arginine biosynthetic enzymes tested was regulated. Comparisons were made between the specific activities of these enzymes in bacteria grown on arginine or on its precursor, glutamate. N2-Acetylornithine 5-aminotransferase (ACOAT), an enzyme involved in both the biosynthesis and catabolism of arginine, was induced about 14-fold during growth of the organism on arginine as the only carbon and nitrogen source, and the anabolic ornithine carbamoyltransferase (aOTC), a strictly biosynthetic enzyme, was repressed 18-fold. Addition of various carbon sources to the arginine medium led to repression of ACOAT and to derepression of aOTC. Fructose, which supported only slow growth of P. aeruginosa, had a weak regulatory effect on the synthesis of the two arginine enzymes while citrate, a good carbon source for this organism, had a strong effect. The repression of ACOAT by citrate was not relieved by adding cyclic AMP to the medium. Under a variety of growth conditions leading to different enzyme activities, a linear relationship between the reciprocal of the specific activity of ACOAT and the specific activity of aOTC was observed. This inverse regulation of the formation of the two enzymes suggested that a single regulatory system governs their synthesis. Such a view was supported by the isolation of citrate-resistant regulatory mutants which constitutively formed ACOAT at the induced level and aOTC at the repressed level.     

PprI和RecX蛋白对耐辐射奇球菌抗氧化作用的影响

利用基因突变、化学发光法和酶活性分析研究了耐辐射奇球菌中与辐射抗性密切相关的基因pprI(Dr0167)和recX(Dr1310)突变对菌体活性氧清除作用的影响,分析了其对抗氧化酶活性的调控功能。实验结果表明,缺失pprI的突变株对活性氧自由基氧化异常敏感,过氧化氢酶和超氧化物歧化酶活性显著降低。与之相反,RecX对菌体活性氧清除作用表现为一种“负”的影响,即缺失recX的突变株对活性氧自由基的清除能力反而增强了,过氧化氢酶和超氧化物歧化酶的酶活性明显增加。表明这两个基因与抗氧化系统的调控有关。为进一步研究该菌的抗氧化机制提供了一些思路。     

Development of the activities of oxidative, glycolytic and muscle enzymes during early larval life in three families of freshwater fish

The development of the activities of oxidative (COX, CS), glycolytic (PFK, PK, LDH) and muscle enzymes (CK, MK, Pase) was studied in representatives of the families Coregonidae, Salmonidae and Cyprinidae, from hatching to an age of approximately 100 days. In addition, the activities of two enzymes of amino acid metabolism (GOT, GPT) were followed in rainbow trout and in roach.
Water content of fresh body weight and protein content of dry body weight decrease during the early larval period. Specific activities of the two oxidative enzymes decline, whereas those of glycolytic and muscle enzymes increase in all species.
A family-specific event is the enormous increase in glycolytic and muscle enzymes from very low values in the early larva to very high levels in adult Coregonus sp. In rainbow trout, CS activity begins with a low-level period lasting throughout the yolk-sac period, whereas in the other species CS activity is high immediately after hatching.
Acclimation to either 15 or 20° C has no effect on the mass-specific activities of PFK, M K, CK and Pase in roach and chub, but the former three enzymes appear to be strongly dependent on rearing conditions during the early larval period, whereas Pase is not.     

Effect of lanthanum ions (La3+) on the reactive oxygen species scavenging enzymes in wheat leaves

Physiological effects of lanthanum ions on the activities of the enzymes in the reactive oxygen species (ROS) scavenging system in leaves of wheat (Triticum aestivum L.) seedlings were studied. Wheat leaves treated in Hogland solution with 0.1 mM LaCl₃ for 48 h showed increased levels of superoxide dismutase (SOD), catalase (CAT), ascorbate-specific peroxidase (AsA-POD), and dehydroascorbate reductase (DHAR). However, a minor effect was observed on the levels of monodehydroascorbate reductase (MDAR) and glutathione reductase (GR), which regulate the release of energy required by the ROS scavenging system. The whole system was linked up by H⁺ transmission. Our results indicated that the activities of the enzymes that function directly to remove ROS were elevated by La³⁺ treatment, which is consistent with the observations that La³⁺-treated plants had increased tolerance to environmental stresses. The remaining levels of MDAR and GR suggested that these two enzymes might be regulated differently from that of the other four enzymes studied.     

Enzymes of the purine interconversion system in chronic lymphatic leukemia: decreased purine nucleoside phosphorylase and adenosine deaminase activity.

Activities of adenosine deaminase (ADA), adenosine kinase (AK), adenine phosphoribosyltransferase (APRT), hypoxanthine guanine phosphoribosyltransferase (HGPRT), and purine nucleoside phosphorylase (PNP), all enzymes of the purine interconversion system, were determined in lymphocytes of 25 patients with chronic lymphatic leukemia (CLL) and in 23 controls. A statistically significant decrease of PNP activities and a reduction of ADA activities at borderline levels were found in the patients, whereas for the other enzymes assayed no deviation from normal values was observed.     

近地层臭氧对小麦抗氧化酶活性变化动态的影响

摘要:利用开顶式气室（TOCs）模拟了地表O3浓度升高的大田试验条件,测定了小麦各生长期(返青期、拔节期、孕穗期、抽穗期、灌浆期、成熟期)叶片抗氧化酶的活性。通过两年的重复试验,得到相似结果。试验表明,过氧化氢酶（CAT）和过氧化物酶（POD）活性在小麦的生长进程中,先升高后降低。O3浓度升高使CAT酶和POD酶活性的峰值期提前,POD酶峰值期明显提前所需的O3浓度比CAT酶高,O3诱导小麦生长前期的CAT、POD酶活性升高,后期则起抑制作用。POD酶对低浓度O3的胁迫响应时间较晚。当O3浓度提高到150 nL?L-1时,超氧化物歧化酶（SOD）活性随小麦生长期的变化,由原来的双峰曲线变为单峰曲线,高浓度O3使SOD酶活性在抽穗期后持续显著降低。O3不明显改变抗坏血酸过氧化物酶（APX）对胁迫的响应时间,但一定浓度范围内O3能诱导APX酶活性升高。不同浓度的O3对SOD酶和APX酶的作用效应和强度显著不同, CAT酶和POD酶对O3比较敏感。 因此,近地层O3浓度增加,改变了CAT、POD和SOD酶活性变化的时序特征,抗氧化酶对O3胁迫的响应特征取决于小麦的生育期、O3的浓度和薰气时间。     

Hypolipidemic drug clofibrate induces hepatic dedifferentiation

The effect of clofibrate on rat liver enzymes and metabolites was compared with that produced by partial hepatectomy and an extrahepatic tumor. Clofibrate administration produced decrease in gamma-glutamyltranspeptidase (GGT) activity with concomitant increase in glutathione concentration. The drug was able to exert its GGT-lowering effect even when fed to tumor-bearing animals. Presence of an extrahepatic neoplasm as well as administration of clofibrate resulted in marked decrease in activities of hepatic arginase and ornithine transaminase. Administration of clofibrate to the tumor-bearing rat produced a further decrease in activities of these two enzymes. These results suggest that clofibrate causes hepatic dedifferentiation and simulates an extrahepatic tumor. However, clofibrate did not induce any significant increase in polyamine profile unlike the other two experimental conditions.