砷的生物转化与代谢机制研究进展

砷广泛分布于自然界中,与人类的活动与健康息息相关.一方面,砷中毒与砷污染事件在全球范围内的频发,严重威胁人类的健康.目前的研究发现自然界中的抗砷微生物直接参与了砷的地球化学循环,在砷的生物转化以及在砷污染的生物治理中起到重要的作用.另一方面,砷作为毒物的同时亦是一种有效的中西药制剂,在治疗血液系统、恶性淋巴系统疾病及癌症中具有明显的疗效,因此砷在人体的转化过程与机制研究有待深入的探讨.有鉴于此,对砷在自然界和人体中的转化形式以及抗砷微生物的抗砷机制进行了阐述,并对砷的微生物转化在环境与医学中的应用进行了展望.     

克雷伯杆菌甘油脱氢酶和1,3-丙二醇氧化还原酶的动力学机制

本研究主要对克雷伯杆菌甘油转化1,3-丙二醇代谢途径中的2个关键酶甘油脱氢酶(GDH)、1,3-丙二醇氧化还原酶(PDOR)反应机制和动力学进行了研究。首先,通过初速度和产物抑制动力学研究确定了GDH、PDOR双底物酶促反应机制为有序BiBi机制,明确了由反应物消耗到产物生成之间的历程。其次,建立了GDH、PDOR双底物酶促反应动力学模型,由动力学模型可知,在偶合反应中,如果GDH和PDOR酶量相同,GDH氧化反应成为限速反应,而辅酶I将主要以氧化型NAD+形式存在。动力学信息为酶法合成1,3-丙二醇和代谢工程研究提供理论指导。     

紫色非硫细菌的砷代谢机制

[目的]系统阐述紫色非硫细菌(PNSB)砷代谢机制和砷代谢基因簇的进化关系.[方法]通过生物信息学方法分析了PNSB砷代谢基因簇的分布、组成、排布方式.采用UV-Vis和HPLC-ICP-MS方法,研究了3个PNSB种类对砷的抗性、砷形态及价态的转化、砷在细胞中的积累和分布以及磷酸盐对As细胞毒性的影响.[结果]砷基因簇分析表明:已公布全基因组序列的17个PNSB菌株基因组中均含有以ars operon为核心的砷代谢基因簇,由1-4个操纵子组成,主要含有与细胞质砷还原和砷甲基化代谢相关的基因,但基因的组成和排列方式因种和菌株而异,尤其是arsM和两类进化来源不同的arsC.实验结果表明:光照厌氧条件下,3个PNSB种类对As(V)和As(Ⅲ)均具有抗性,As(V)和As(Ⅲ)均能进入细胞 ;在胞内As(V)能够还原为As(Ⅲ)并被排出胞外,但不能将As(Ⅲ)氧化为As(V),也未检测到甲基砷化物 ;磷酸盐浓度升高,能够抑制As(V)进入细胞,降低As(V)对细胞的毒性,而不能抑制As(Ⅲ)进入细胞.[结论]PNSB砷代谢机制主体为细胞质As(V)还原,也还有砷甲基化途径.通过对砷代谢基因簇结构多样性特点和进化方式分析,提出了与Rosen不同的ars operon进化途径.这对深入开展PNSB砷代谢和基因之间的相互作用研究奠定基础.     

砷的微生物转化及其在环境与医学应用中的研究进展

砷广泛分布于自然界中,近年来砷污染及砷中毒事件在全球范围内的频繁发生,严重威胁着全球上千万人口的健康.砷的迁移与转化等地球化学行为的研究对探明环境中砷的来源以及砷污染整治的方法至关重要.越来越多的研究表明,自然界中的微生物广泛参与了砷的地球化学循环,在砷的迁移与转化过程中起到关键作用.综述了近年来砷的微生物转化及其相关酶类与编码基因等的研究进展,同时结合作者的工作分析与展望了砷的微生物转化在环境与医学中的应用前景.     

认识不可逆抑制酶促反应

认识不可逆抑制酶促反应任维栋,张桂春（潍坊医学院生化教研室,山东潍坊２６１０４２）关键词不可逆抑制作用,酶,动力学不可逆抑制剂对研究酶的活性中心、药物毒物的作用机理及物质代谢途径与调节起着重要作用。１９６５年,邹承鲁发表了不可逆抑制酶促反应动力学的论...     

抑制剂对有机相酶促己酸乙酯合成中固定化脂肪酶影响

稳定的催化活性和选择性的调节与控制已成为研究有机相酶促反应机制和应用的重要内容。由于选择和优化反应条件的方法具有较大局限性．目前有机介质中酶催化选择性和稳定性的调节与控制的研究除了常用的固定化手段外,更关注通过改变酶分子自身的一些方法上。如蛋白质工程、酶的化学修饰和非共价修饰〔1.2〕。相比之下,其中非共价修饰具有方便、实用的特点。酶的修饰剂大多也是酶的抑制剂。抑制剂已用来研究脂肪酶的结构与代谢特征〔3〕。Russel,Guo等通过其改变酶的构象和界面特征来调节和控制酶稳定的特异性〔4.5〕。我们在对微生物脂肪酶正庚烷中合成短链芳香酯研究基础上〔6.7〕。本文报道用具有两亲特性的表面活性剂、胆汁盐和金属离子这些脂肪酶的抑制剂对庚烷中脂肪酶酶促己酸乙酯酯化反应的影响,以促进酶活性的调节和控制的研究和应用。     

中国莲（Nelumbo nucifera）幼苗抗氧化系统对砷胁迫的响应

为了研究中国莲（Nelumbo nucifera）抗氧化系统对砷胁迫的响应,研究比较了两种不同价态无机砷As（Ⅲ）和砷As（V）对中国莲幼苗可溶性蛋白含量、丙二醛（MDA）含量以及抗氧化系统的影响。结果表明,中国莲幼苗中MDA和可溶性蛋白质含量随砷浓度的增加呈现先升高后降低的趋势。中国莲幼苗的MDA和蛋白质含量受As（V）的影响不如As（11]）敏感。抗氧化系统酶中,超氧化物歧化酶（SOD）对砷处理最敏感,当As（11I）浓度在2．5pmol／L和As（V）浓度在100μmol／L时,SOD酶活性显著高于对照组。过氧化物酶（POD）在As（Ⅲ）处理浓度为10umol／L时就出现显著上升,相对而言,过氧化氢酶（CAT）对As（V）比较敏感。实验结果表明,随着浓度的增加,砷对幼苗产生的氧化胁迫导致SOD、CAT和POD三种酶活性有所增加,以配合清除细胞内的活性氧自由基（ROS）,维持细胞代谢的稳定。本研究为进一步研究砷胁迫下莲的生理和生长变化、以及莲的培育和移植提供了部分基础数据。     

砷对大鼠生精细胞凋亡的影响

砷是已肯定的生殖毒物之一,但关于砷对雄（男）性生殖系统的毒性作用的分子机制尚不清楚。本实验通过检测慢性砷中毒大鼠睾丸脏器系数、精子头数、每日精子生成量（DSP）及生精细胞凋亡情况,探讨砷对精子发生的影响,为进一步探讨砷致雄（男）性生殖毒性机制提供基础资料。结果显示,随染砷剂量的增加,大鼠出现睾丸生精上皮结构变疏松,生精上皮细胞层次紊乱,甚至部分生精小管基膜溶解、管腔中成熟精子减少、小管间隙增宽、细胞体积缩小、间质出现水肿、渗出等病理改变;睾丸精子头计数及DSP逐渐减少,中、高剂量组变化尤其明显。     

氮磷比对有机磷环境中微藻砷代谢的生态风险效应北大核心

【目的】探究了以单酯磷D-葡萄糖-6-磷酸二钠(D-glucose-6-disodium phosphate,GP)为唯一磷源时含砷(As5+)水体中不同氮磷质量比对铜绿微囊藻(Microcystis aeruginosa)生长及砷代谢和微囊藻毒素(microcystins,MCs)释出的影响。【方法】将氮磷饥饿状态的藻细胞于含砷水体中不同氮磷比条件下进行实验,通过测定藻细胞密度(OD680)、叶绿素a(chlorophyll a,Chla)、实际光合产率(Yield)、超氧化物歧化酶(superoxide dismutase,SOD)、砷的存在形态以及微囊藻毒素含量,分析该藻在砷胁迫下的生理响应以及砷代谢机制。【结果】氮磷饥饿状态藻细胞对较高GP水平(0.1 mg/L)下的低氮磷比有更好的适应性,较低GP水平(0.02 mg/L)下的高氮磷比能显著促进培养初期藻细胞的OD680、Chla和Yield;SOD在培养初期与末期受氮磷比影响显著。GP环境下铜绿微囊藻经8 d培养后氮磷比为10:0.1介质中的砷表现为以亚砷酸盐(As^(3+))为主,占水体总砷(total arsenic,TAs)含量的78.8%,其余氮磷比环境中仍以As^(5+)为主,藻体砷形态则均以As^(5+)为主,氮磷比为1:0.1时有机砷占藻体TAs比例最高。藻细胞砷代谢受GP水平影响显著,较高GP环境(0.1 mg/L)下砷的代谢总量也更高,氮磷比为10:0.1时砷代谢以As^(5+)的还原和As^(3+)释出为主,低GP环境下(0.02 mg/L)砷代谢的甲基化水平提高。介质中MCs的含量与GP水平有关,较高GP的低氮磷比水体中MCs含量最低。【结论】研究结果对全面了解有机磷源含砷水体中藻华暴发及砷生态风险的科学管控具有重要意义。     

代谢的调节

概况生物的生存依赖于有机体不停地进行物质代谢。食进的有机营养物质主要有糖、脂及蛋白质三大类;主要的代谢器官和组织有肝、肾、肌肉及脂肪组织;主要的排泄物有H_2O、CO_2及尿素。体内各物质的生成、分解以及功能的体现,包括数目繁多的酶促反应。完整的物质代谢过程是由很多酶促反应以一定的顺序连续进     

Oxidative stress by inorganic arsenic: modulation by thyroid hormones in rat

Arsenic toxicity is attributed mainly to lipid peroxidation and oxidative stress. We therefore studied the modulatory effects of thyroid hormones on arsenic toxicity in rat on lipid peroxidation and oxidative stress. Thyroid hormones, through a mechanism unknown at present, inhibit arsenic accumulation in liver and kidney. Mobilization of arsenic apparently diminishes lipid peroxidation and improves reduced glutathione status, two biochemical demands of combating arsenic toxicity. Results are discussed in reference to the effect of thyroid hormones on microsomal metabolism of arsenic. Arsenic is less toxic in hyperthyroid than in hypothyroid rats. A physiological antagonism between arsenic and thyroxine is discussed.     

Arsenic in contaminated waters: Biogeochemical cycle, microbial metabolism and biotreatment processes

Arsenic is responsible for the contamination of water supplies in various parts of the world and poses a major risk to human health. Its toxicity and bioavailability depend on its speciation, which in turn, depends on microbial transformations, including reduction, oxidation and methylation. This review describes the development of bioprocesses for the treatment of arsenic-contaminated waters based on bacterial metabolism and biogeochemical cycling of arsenic.     

Microbial arsenic: from geocycles to genes and enzymes

Arsenic compounds have been abundant at near toxic levels in the environment since the origin of life. In response, microbes have evolved mechanisms for arsenic resistance and enzymes that oxidize As(III) to As(V) or reduce As(V) to As(III). Formation and degradation of organoarsenicals, for example methylarsenic compounds, occur. There is a global arsenic geocycle, where microbial metabolism and mobilization (or immobilization) are important processes. Recent progress in studies of the ars operon (conferring resistance to As(III) and As(V)) in many bacterial types (and related systems in Archaea and yeast) and new understanding of arsenite oxidation and arsenate reduction by respiratory-chain-linked enzyme complexes has been substantial. The DNA sequencing and protein crystal structures have established the convergent evolution of three classes of arsenate reductases (that is classes of arsenate reductases are not of common evolutionary origin). Proposed reaction mechanisms in each case involve three cysteine thiols and S-As bond intermediates, so convergent evolution to similar mechanisms has taken place.     

Arsenic Carcinogenesis in the Skin

Summary Chronic arsenic poisoning is a world public health issue. Long-term exposure to inorganic arsenic (As) from drinking water has been documented to induce cancers in lung, urinary bladder, kidney, liver and skin in a dose–response relationship. Oxidative stress, chromosomal abnormality and altered growth factors are possible modes of action in arsenic carcinogenesis. Arsenic tends to accumulate in the skin. Skin hyperpigmentation and hyperkeratosis have long been known to be the hallmark signs of chronic As exposure. There are significant associations between these dermatological lesions and risk of skin cancer. The most common arsenic-induced skin cancers are Bowen’s disease (carcinoma in situ), basal cell carcinoma (BCC) and squamous cell carcinoma (SCC). Arsenic-induced Bowen’s disease (As-BD) is able to transform into invasive BCC and SCC. Individuals with As-BD are considered for more aggressive cancer screening in the lung and urinary bladder. As-BD provides an excellent model for studying the early stages of chemical carcinogenesis in human beings. Arsenic exposure is associated with G2/M cell cycle arrest and DNA aneuploidy in both cultured keratinocytes and As-BD lesions. These cellular abnormalities relate to the p53 dysfunction induced by arsenic. The characteristic clinical figures of arsenic-induced skin cancer are: (i) occurrence on sun-protected areas of the body; (ii) multiple and recrudescent lesions. Both As and UVB are able to induce skin cancer. Arsenic treatment enhances the cytotoxicity, mutagenicity and clastogenicity of UV in mammalian cells. Both As and UVB induce apoptosis in keratinocytes by caspase-9 and caspase-8 signaling, respectively. Combined UVB and As treatments resulted in the antiproliferative and proapoptotic effects by stimulating both caspase pathways in the keratinocytes. UVB irradiation inhibited mutant p53 and ki-67 expression, as well as increased in the number of apoptotic cells in As-BD lesions which resulted in an inhibitory effect on proliferation. As-UVB interaction provides a reasonable explanation for the rare occurrences of arsenical cancer in the sun-exposed skin. The multiple and recurrent skin lesions are associated with cellular immune dysfunction in chronic arsenism. A decrease in peripheral CD4+ cells was noticed in the inhabitants of arsenic exposure areas. There was a decrease in the number of Langerhans cells in As-BD lesion which results in an impaired immune function on the lesional sites. Since CD4+ cells are the target cell affected by As, the interaction between CD4+ cells and epidermal keratinocytes under As affection might be closely linked to the pathogenesis of multiple occurrence of arsenic-induced skin cancer. In this review, we provide and discuss the pathomechanisms of arsenic skin cancer and the relationship to its characteristic figures. Such information is critical for understanding the molecular mechanism for arsenic carcinogenesis in other internal organs.     

Arsenic and weight loss: At a crossroad between lipogenesis and lipolysis

Arsenic is found in soil, food, water and earth crust. Arsenic exposure is associated with chronic diseases such as cancer, cardiovascular disease as well as diabetes. One of complex effects of arsenic is on weight gain or loss. Involvement of arsenic in both weight loss and gain signaling pathways has previously been reported; however, too little attention has been paid to its weight reducing effect. Animal studies exhibited a role of arsenic in weight loss. In this regard, arsenic interference with endocrine system, leptin and adiponectin hormones as well as thermogenesis is more evidence. Apparently, arsenic-induced weight lossis generally meditated by its interaction with thermogenesis. In this review we have discussed the irregularities in metabolic pathways induced by arsenic that can lead to weight loss.     

Enumeration and characterization of culturable arsenate resistant bacteria in a large estuary

Arsenic is a toxic element that exists in two major inorganic forms, arsenate and arsenite. A number of bacteria have been shown to resist arsenic exposure, and even more bacteria appear to possess the genes for arsenic resistance. In this study, the numbers of culturable arsenate-resistant bacteria present in water at three coastal sites in the Lake Pontchartrain estuary, Louisiana, was determined. Despite insignificant (less than 1.33 μM) levels of arsenic in this system, 20–50% of the viable count of bacteria showed appreciable arsenate resistance, suggesting that arsenic-resistant bacteria are common and widespread. A diverse array of arsenate-resistant isolates was obtained, with 16S rRNA sequence analysis indicating 37 different bacterial strains, representing six major bacterial groups. Many of these isolates were affiliated with groups of bacteria that have been poorly characterized in terms of arsenic resistance, such as the Betaproteobacteria or Flavobacteria. Some isolates were capable of tolerating very high (>100 mM) levels of arsenate, although arsenite resistance was generally much lower. The results suggest that arsenic-resistant bacteria are common, even in environments with insignificant arsenic contamination, and that many different groups of aquatic bacteria show appreciable arsenic resistance.     

High arsenic intake raises kidney copper and lowers plasma copper concentrations in rats

The effect of high arsenic intake on copper metabolism was investigated. Male rats aged 6 wk had free access to purified diets containing either 0 or 100 mg As/kg diet and demineralized water for a period of 2 wk. Arsenic was added to the diet in the form of NaAsO₂. The high-arsenic diet decreased feed and water intake and body weight gain, but significantly increased liver weight. Kidney weight was not affected. Arsenic feeding drastically elevated kidney copper concentration, but significantly reduced copper concentration in plasma. Both true absorption and biliary excretion of copper were decreased significantly in rats fed the high-arsenic diet. True copper absorption was lowered essentially through the lower copper intake in the rats fed arsenic. It is speculated that arsenic feeding primarily leads to copper accumulation in the kidney, followed by a decrease in feed intake and thus in true, absolute copper absorption, a decrease in plasma copper concentration, and a decrease in biliary copper excretion.     

Potential molecular mechanisms for combined toxicity of arsenic and alcohol

Arsenic is a ubiquitous environmental factor that has been identified as a risk factor for a wide range of human diseases. Alcohol is clearly a toxic substance when consumed in excess. Alcohol abuse results in a variety of pathological effects, including damages to liver, heart, and brain, as well as other organs, and is associated with an increased risk of certain types of cancers. In history, arsenic-contaminated beers caused severe diseases. There are populations who are exposed to relatively high levels of arsenic in their drinking water and consume alcohol at the same time. In this focused review, we aim to discuss important molecular mechanisms responsible for arsenic toxicity and potential combined toxic effects of alcohol and arsenic.     

砷对烤烟碳氮代谢及其产量和品质的影响

采用盆栽试验,系统地研究了砷对烤烟全生育期的碳氮代谢及其产量和品质的影响。结果表明,砷毒害对烤烟全生育期的碳代谢有显著影响,抑制了碳的同化和转化,降低了整个生育期的叶绿素含量、光合速率,造成了全生育期可溶性糖的积累,导致了生育后期淀粉含量的降低,最终使碳积累减少。砷毒害也改变了烤烟的氮代谢,造成生育前期氮同化能力的降低,表现出硝酸还原酶(NR)活性下降、总氮和蛋白质含量低于CK。砷毒害烤烟的氮转化表现活跃,提高了其中的游离氨基酸含量和谷氨酸-丙酮酸转氨酶(GPT)活性,最终导致烤烟生育中后期总氮和蛋白质的积累,但使整个生育期的烟碱含量降低。研究还表明,砷毒害降低了烤烟的产量和经济性状,增加了叶片中砷的积累,可溶性总糖含量的提高和糖氮比的协调虽好,但烟碱含量的降低和总氮、蛋白质含量的增加,以及糖碱比和氮碱比的失调,不利于碳氮代谢有关的化学品质形成。