重金属镉对鲫鱼碱性磷酸酶和酸性磷酸酶活性的影响

研究了重金属镉对鲫鱼肠、肝胰脏、鳃组织碱性磷酸酶和酸性磷酸酶活性的影响。结果表明,在0.2、0.4、0.8mg/L镉浓度条件下静态染毒12h、24h、48h、96h后,鲫鱼肠、鳃组织中碱性磷酸酶(AKP)和酸性磷酸酶(ACP)的活性降低,肝和胰脏的碱性磷酸酶活性没有明显变化,其酸性磷酸酶活性则升高。     

高效氯氰菊酯对草鱼器官酸性磷酸酶活性的影响

研究了暴露于不同高效氯氰菊酯浓度下草鱼Ctenopharyngodon idellas鳃、肝胰脏和肾脏的酸性磷酸酶(Acid Phosphatase,ACP)的活性变化。实验中高效氯氰菊酯浓度设5组,其中对照组1个(0μg/L),处理组4个(0.5μg/L、1.0μg/L、3.0μg/L、5.0μg/L)。每组随机投放40尾草鱼,分别于1d、5d、12d取样,测定鳃、肝胰脏和肾脏的ACP活性。结果显示,鳃ACP活性低浓度(0.5μg/L、1.0μg/L)短时间(1d)暴露下显著增加(P<0.01),高浓度(3.0μg/L、5.0μg/L)暴露时显著下降(P<0.01);肝胰脏ACP活性暴露1d时,除0.5μg/L组外,其他处理组均显著下降(P<0.05、P<0.01),暴露5d、12d时,各处理组均显著上升(P<0.05、P<0.01);肾脏ACP活性除0.5μg/L组在暴露1d、5d时无显著变化(P>0.05)外,其他处理组在暴露时间内均显著下降(P<0.01)。实验表明:高效氯氰菊酯能够通过影响草鱼鳃、肝胰脏和肾脏中酶的活性,干扰这些器官的代谢平衡,进而对这些器官产生毒害作用。     

重金属铅对鲫鱼乳酸脱氢酶和过氧化氢酶活性的影响

在实验室条件下,采用毒性实验方法研究不同浓度重金属铅(Pb2 )对鲫鱼血清乳酸脱氢酶(LDH)和血液过氧化氢酶(CAT)活性的影响.研究结果表明,随着金属离子铅浓度的增高血清中乳酸脱氢酶和过氧化氢酶活性下降,但两种酶对铅离子影响的敏感程度不同.影响LDH活性的最低铅离子浓度为0.1mmol/L,而CAT为0.5mmol/L,当Pb2 浓度为1mmol/L时,LDH活性降至10%左右,而CAT仅下降50%左右.对重金属离子浓度与酶活性的定性和定量分析为有效监控鱼类生存环境提供了有价值的参考资料.     

镉和铅对鲫鱼肝胰脏过氧化氢酶活性的影响

采用急性毒性实验方法,研究了不同ρCd2+和ρPb2+及二者混合溶液,染毒24、48、72、96 h后,对鲫鱼肝胰脏过氧化氢酶(CAT)活性的影响.结果表明:在实验剂量范围(ρCd2+为0.5～6.0 mg/L,ρPb2+为10.0～40.0 mg/L),0.5、2.0 mg/L的Cd2+对CAT活性有诱导作用,6.0 mg/L的Cd2+对CAT活性有抑制作用;Pb2+胁迫对CAT活性随着染毒时间的延长而逐渐降低;2.0 mg/L Cd2+与20.0 mg/L Pb2+混合胁迫对CAT活性有明显的诱导作用;0.5mg/L Cd2++10.0 mg/L Pb2+,6.0 mg/L Cd2++40.0 mg/L Pb2+混合胁迫对CAT活性有明显抑制作用.Cd2+和Pb2+胁迫对CAT活性的影响呈剂量效应,二者关系曲线为抛物线,抛物线顶点是鲫鱼对Cd2+和Pb2+污染从适应到中毒反应的阈值,可间接作为检测环境污染情况的指标.     

鲫鱼酸性磷酸酶酶学特性及不同效应物对酶活力的影响

经NaAc-HAc缓冲液(pH5.0)抽提,正丁醇处理,硫酸铵分级沉淀,DEAE-32离子交换层析,SephadexG-150凝胶过滤纯化,从鲫鱼内脏中分离纯化出电泳纯的酸性磷酸酶。该酶提纯倍数为30.82,比活力195.06U/mg。研究表明,该酶催化对硝基苯磷酸二钠水解反应,最适pH4.8,pH小于4和大于7时不稳定;最适温度45℃,温度高于50℃不稳定;米氏常数为0.23mmol/L,利用SDS-PAGE测定酶亚基分子量为33.3kD。化学修饰剂SUAN、PMSF、DTT、NBS对该酶活力影响不大,BrAc和IAc有明显抑制作用。金属离子对该酶催化活力有不同影响,Na+、K+、Ni2+、Co2+影响不显著,Mg2+、Ca2+、Ba2+、Mn2+有激活作用,Ag+、Cu2+、Pb2+、Cd2+有抑制作用,其中Mg2+、Ca2+、Pb2+、Cd2+对鲫鱼酸性磷酸酶荧光光谱的影响表明金属离子对酶活力的影响与酶构象改变有关。       

氦氖激光穴位照射对兔子宫和卵巢酸性磷酸酶（ACP）、碱性磷酸酶（ALP）的作用效应

用不同功率的Ｈｅ—Ｎｅ激光照射雌兔外生殖器,照射穴位,用生化方法分别测定兔子宫和卵巢的酸性磷酸酶（ＡＣＰ）、碱性磷酸酶（ＡＬＰ）的活性。激光照射后,子宫和卵巢的ＡＣＰ酶活性保持基本恒定状态,与正常对照组比较无显著差异;相反,ＡＬＰ酶活性显著提高。分析这两种酶在激光用射后表现出来的差异,作者认为是激光作用使细胞内环境改变之故,且这两种酶对Ｈｅ—Ｎｅ激光的耐受性有所不同。     

电气石处理水对Caco-2细胞生长和碱性磷酸酶活性的影响

水是由若干水分子通过氢键结合形成的水分子团簇结构,水分子团簇的改变能产生多种生物学效应。研究了电气石对液态水团簇的影响,并以Caco-2细胞培养模型,探讨了电气石处理DMEM培养液对细胞生长和碱性磷酸酶活性的影响。研究结果显示：电气石使蒸馏水^17O核磁共振(^17O nucleaur magnetic resonance,^17O NMR)半高幅宽变窄,降低了水分子缔合度;电气石处理的DMEM培养液培养Caco-2细胞促进了细胞生长,提高了细胞碱性磷酸酶活性。结果表明电气石可降低水分子缔合度,促进细胞生长和分化。     

金属离子和脲对白蜡虫碱性磷酸酶的影响

各种金属离子及脲对白蜡虫Ericerus pela (Chavannes)碱性磷酸酶的活性有不同的影响。从白蜡虫雌成虫中分离纯化得到碱性磷酸酶,加入各种不同浓度的金属离子及脲测定酶的活力。一价金属离子Na⁺、K⁺、Li⁺对酶活力没有影响。碱土金属离子Ca²⁺、Mg²⁺、Ba²⁺对酶有激活作用,激活作用的大小顺序依次为Ca²⁺、Ba²⁺、Mg²⁺。第一过渡金属离子中,Mn²⁺、Co²⁺、Ni²⁺对酶有激活作用,而Zn²⁺、Cu²⁺有抑制作用。重金属离子Cd²⁺、Pb²⁺对酶有抑制作用。Ca²⁺激活作用表现为非竞争性激活效应。Cu²⁺抑制作用表现为非竞争性抑制效应。脲对碱性磷酸酶有变性失活作用,按脲浓度可分为低于3 mol/L和高于3 mol/L两种类型。低浓度的脲对白蜡虫碱性磷酸酶的活性抑制的动力学表现为混合型效应。     

部分根系供磷对黄瓜根系和幼苗生长及根系酸性磷酸酶活性影响

通过分根处理研究了部分根系供磷对黄瓜幼苗生长、植株体内的含磷量及根系酸性磷酸酶活性的影响。结果表明 ,2 0 %根系缺磷 (1条根缺磷 ,4条根供磷 )可以促进根系及植株地上部的生长 ,其根系及地上部的生物量分别是正常生长植株的 1.39倍和 1.2 1倍。2 0 %根系缺磷 ,还可以促进其它供磷根系对磷的吸收。分根处理后 ,2 0 %根系缺磷不影响植物对磷营养的需要 ,但却表现出了R/S比增大的典型缺磷反应 ,说明植物感应缺磷根系起着比地上部更为重要的作用。分根处理后不供磷根系的酸性磷酸酶活性显著高于供磷根系的酸性磷酸酶活性 ,并且根系的酸性磷酸酶活性只与根系的含磷量显著相关 ,与地上部的磷营养状况关系不明显。这说明 ,缺磷条件下 ,黄瓜植株根系分泌酸性磷酸酶活性的增高 ,是黄瓜根系对低磷胁迫的适应性机理 ,而不是地上部改善体内磷营养的调控机理。     

模拟氮沉降对鼎湖山森林土壤酸性磷酸单酯酶活性和有效磷含量的影响

采用野外原位试验模拟氮(N)沉降,研究了其对鼎湖山马尾松林、混交林和季风林3种森林类型土壤酸性磷酸单酯酶活性(APA)和有效磷(AP)含量的影响.在季风林中设置对照(0 kg N·hm-2·a-1)、低N(50 kg N·hm-2·a-1)、中N(100 kg N·hm-2·a-1)和高N(150 kg N·hm-2·a-1)处理,在马尾松林和混交林中只设置对照、低N和中N处理.结果表明:随着土层加深,土壤APA和AP含量降低.土壤APA在季风林中最高,而AP含量在3种林型中没有显著差异.N沉降增加对土壤APA的作用与林型有关.季风林中适度N沉降可使APA升高,且低N处理的APA(19.52 μmol·g-1·h-1)最高;马尾松林和混交林中,中N处理的APA最高,分别为12.74和11.02 μmol·g-1·h-1.3种林型的AP含量均在低N处理下最高,但各N处理之间的差异并不显著.土壤APA与AP含量之间呈显著正相关关系.     

非特异性酯酶及酸性和碱性磷酸酶在奥尼罗非鱼消化道的分布

本研究利用石蜡切片结合组织化学方法研究奥尼罗非鱼(Oreochromis niloticus♀×O. aureus♂)消化道内非特异性酯酶、酸性磷酸酶和碱性磷酸酶的分布特点。结果发现,非特异性酯酶大量密集分布在前肠,其次为胃、食管和中肠,极少数零散分布在后肠。酸性磷酸酶主要分布于奥尼罗非鱼前肠,其次为中肠和胃,极少数分布在食管和后肠。碱性磷酸酶大量密集分布在奥尼罗非鱼前肠和中肠,其次为胃、食管和后肠。结果表明,奥尼罗非鱼前肠、中肠、食管和胃有较强的吸收和消化脂质功能,前肠、中肠和胃是蛋白质吸收和消化的主要部位,前肠和中肠是营养物质吸收的主要部位。     

Effects of Magnesium Ions on Thermal Inactivation of Alkaline Phosphatase

The effect of Mg²⁺ on the thermal inactivation and unfolding of calf intestinal alkaline phosphatase has been studied at different temperatures and Mg²⁺ concentrations. Increasing the Mg²⁺ concentration in the denatured system significantly enhanced the inactivation and unfolding of the enzyme during thermal inactivation. The analysis of the kinetic course of substrate reaction during thermal inactivation showed that at 47°C the increased free Mg²⁺ concentration caused the inactivation rate to increase. Increasing the temperature strengthened the effect of Mg²⁺ on the thermal inactivation. Control experiment showed that this is not due to salt effect. The time course of fluorescence emission spectra showed that the emission maximum for Mg²⁺-containing system was always higher than that of Mg²⁺-free system, and the higher temperature enhanced this difference. In addition, Mg²⁺also enhanced the unfolding rate of the enzyme at 47°C. The potential biological significance of these results are discussed.     

金属离子对牛小肠碱性磷酸酶的影响

经Tris-HCl缓冲液抽提,硫酸铵分级分离沉淀,2次DEAE-32柱层析和Sephadex G-150凝胶过滤,从牛小肠中得到碱性磷酸酶(ALP),提纯倍数为50.69倍,比活为48.87 U/mg,酶液经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)检测呈现单一条带.该酶催化底物对硝基苯磷酸二钠(pNPP)水解反应的最适pH值为9.7,最适温度为45℃.研究了多种金属离子对牛小肠碱性磷酸酶活性的影响,结果表明:一价金属离子Na+、K+对酶活力没有影响;不同的二价金属离子对酶的影响不同,过渡态金属离子中,Ni2+、Co2+对酶的活力影响不大,Mg2+、Ca2+、 Mn2+对酶有不同程度的激活作用,Zn2+、Cu2+对酶有抑制作用;重金属离子Pb2+、Cd2+对酶的活力起抑制作用.     

Alkaline and Acid Phosphatase Demonstration in Human Bone and Cartilage: Effects of Fixation Interval and Methacrylate Embedments

Human bone and cartilage specimens were evaluated for acid and alkaline phosphatase localization following varying fixation periods for fresh or frozen tissue. Formalin fixations of up to 183 hr were followed by embedment in methyl methacrylate; frozen tissue was examined either without fixation or following fixation for up to 1 hr and subsequent glycol or methyl methacrylate embedding. The humeral epiphysis of a young patient with osteogenic sarcoma showed optimum acid and alkaline phosphatase localization following fixation for periods up to 15 hr and embedding in methyl methacrylate. Frozen costochondral junction from a newborn with osteogenesis imperfecta type II showed optimum acid and alkaline phosphatase localization following 30 min fixation in formalin and embedding in methyl methacrylate or after 5 min fixation and embedding in glycol methacrylate.     

Alkaline Phosphatase Content and the Effects of Prednisolone on Mammalian Cells in Culture

The alkaline phosphatase content of different tissue culture cell lines has been shown to vary from no detectable activity to high enzyme concentration. Within the epithelial lines studied alkaline phosphatase is either constitutive or inducible. Two epithelial cell strains in which alkaline phosphatase was "absent" could be induced to develop significant amounts of the enzyme when grown in the presence of Δ¹-hydrocortisone. Phosphate did not repress enzyme induction by prednisolone. Under conditions of deadaptation the induced enzyme was diluted by cell multiplication. The mouse fibroblastic L line and several human fibroblastic lines did not contain alkaline phosphatase when grown under the conditions described nor could they be induced to produce the enzyme when cultivated in medium with prednisolone. Δ¹-Hydrocortisone has other characteristic effects on established mammalian cell cultures which vary among cell lines. Human epithelial lines show reduction in cell multiplication with increase in mitotic index. The cytoplasm is increased and cell volume is nearly doubled. Mouse fibroblasts show a similar reduction in cell multiplication with a decrease in mitotic index. There is no increase in cell cytoplasm. Human fibroblast strains show no inhibition of multiplication or alteration in total cell protein when grown in medium containing prednisolone. Antisera prepared against "negative" prednisolone-inducible human cell lines and against a positive human line inhibited alkaline phosphatase activity to an equal degree.     

Effects of Alkaline Phosphatase Activity on Nucleotide Measurements in Aquatic Microbial Communities

Alkaline phosphatase (APase) activity was detected in aquatic microbial assemblages from the subtropics to Antarctica. The occurrence of APase in environmental nucleotide extracts was shown to significantly affect the measured concentrations of cellular nucleotides (adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, guanosine triphosphate, uridine triphosphate, and cytidine triphosphate), adenylate energy charge, and guanosine triphosphate/adenosine triphosphate ratios, when conventional methods of nucleotide extraction were employed. Under the reaction conditions specified in this report, the initial rate of hydrolysis of adenosine triphosphate was directly proportional to the activity of APase in the sample extracts and consequently can be used as a sensitive measure of APase activity. A method was devised for obtaining reliable nucleotide measurements in naturally occurring microbial populations containing elevated levels of APase activity. The metabolic significance of APase activity in microbial cells is discussed, and it is concluded that the occurrence and regulation of APase in nature is dependent upon microscale inorganic phosphate limitation of the autochthonous microbial communities.     

Effects of Polyethylene Glycol on Bovine Intestine Alkaline Phosphatase Activity and Stability

In this study, we evaluated the effects of polyethylene glycol (PEG) on bovine intestine alkaline phosphatase (BIALP) activity and stability. In the hydrolysis of p-nitrophenylphosphate (pNPP) at pH 9.8 at 20 °C, the k _cat?K _m values of BIALP plus 5–15% w/v free PEG with molecular masses of 1, 2, 6, and 20 kDa (PEG1000, PEG2000, PEG6000, and PEG20000 respectively) were 120–140%, 180–300%, 130–170%, and 110–140% respectively of that of BIALP without free PEG (1.8 μM^?1 s^?1), indicating that activation by PEG2000 was the highest. Unmodified BIALP plus 5% PEG2000 and BIALP pegylated with 2,4-bis(O-methoxypolyethylene glycol)-6-chloro-s-triazine exhibited 1.3-fold higher activity on average than that of BIALP without free PEG under various conditions, including pH 7.0–10.0 and 20–65 °C. The temperatures reducing initial activity by 50% in 30-min incubation of unmodified BIALP plus 5% PEG2000 and pegylated BIALP were 51 and 47 °C respectively, similar to that of BIALP without free PEG (49 °C). These results indicate that the addition of PEG2000 and pegylation increase BIALP activity without affecting its stability, suggesting that they can be used in enzyme immunoassay with BIALP to increase sensitivity and rapidity.     

Chronic Cadmium Exposure Lead to Inhibition of Serum and Hepatic Alkaline Phosphatase Activity in Wistar Rats

Alkaline phosphatase (ALP) activity in the serum and liver from rats administered with cadmium (Cd) in drinking water was studied. After metal administration, Cd showed a time‐dependent accumulation in the liver, meanwhile metallothionein had a maximum increase at 1 month, remaining in this level until the end of the study. On the other hand, serum and liver ALP activity was decreased after 3 months exposure. To determine if Cd produced an inhibition on enzyme, apo‐ALP prepared from both nonexposed and exposed rats was reactivated with Zn, showing 60% more activity as compared with the enzyme isolated from nonexposed rats. In vitro assays showed that Cd‐ALP was partially reactivated with Zn; however, in the presence of cadmium, Zn‐ALP was completely inhibited. Kinetic studies indicate a noncompetitive inhibition by Cd; these results suggest that Cd can substitute Zn, and/or Cd can interact with nucleophilic ligands essential for the enzymatic activity.