基于宏基因组技术的聚磷菌研究进展

聚磷菌是一类非常重要的工程菌,广泛应用于污水处理厂生物除磷过程。聚磷菌可以吸收超过自身所能利用的数倍的磷,在体内合成聚磷化合物从而达到生物除磷的目的。在过去的几年里,宏基因组学以及测序技术的发展大大推动了对聚磷菌物种组成及其磷代谢过程的认识。本文主要对宏基因组技术进行介绍并对近几年基于宏基因组技术深入研究聚磷菌的文章进行综述,以期对这类重要的微生物的生理功能、代谢途径和物种多样性进行全面的了解。     

反硝化聚磷菌的脱氮除磷机制及其在废水处理中的应用

摘要:水体富营养化是当前水污染治理的重点关注对象,利用微生物脱氮除磷开展富营养化水体治理是一种重要的技术。基于反硝化细菌和聚磷菌的脱氮除磷功能,兼具反硝化和聚磷功能的微生物研究及其在污水工艺中的应用越来越广泛。在厌氧和好氧/缺氧环境中,反硝化聚磷菌的脱氮除磷机制有很大差别,且在化学和酶学方面都有所体现。其中,质子驱动力/电子受体理论能够很好地解释反硝化聚磷的化学过程,而反硝化酶系和多聚磷酸盐激酶是酶学过程的主要参与者。当前研究已明确在不同氧含量环境中氮素对磷去除的影响机制,但是否存在磷对除氮作用的影响仍有待进一步研究。在此基础上,本文以氮-磷的偶联过程为切入点,分别从反硝化聚磷的化学过程和酶学机制方面进行简要综述。此外,介绍了反硝化聚磷菌在实验室以及工厂化污水处理中的应用近况,并提出了今后的研究重点与方向,以期为反硝化聚磷菌在环境修复中的进一步开发应用提供理论依据。     

多聚磷酸相关蛋白结构及生物学功能

多聚磷酸（polyphosphate,polyP）是由几个到数百个磷酸基通过高能磷酸酐键连接而成的链状多聚体,存在于所有细胞生物中.多聚磷酸相关蛋白包括多聚磷酸相关酶和多聚磷酸结合蛋白.多聚磷酸相关酶如多聚磷酸激酶（polyphosphate kinase,PPK）催化polyPn生成polyPn+1的可逆反应;外切聚磷酸酶（exopolyphosphatase,PPX）、内切聚磷酸酶（endopolyphosphatase,PPN）能将polyP水解成磷酸残基;多聚磷酸依赖的激酶将polyP的磷转移到生物小分子上,如葡萄糖和烟酰胺腺嘌呤二核苷酸（nicotinamide adenine dinucleotide,NAD）,使其分别磷酸化为6 磷酸葡萄糖和烟酰胺腺嘌呤二核苷酸磷酸（nicotinamide adenine dinucleotide phosphate,NADP）.多聚磷酸结合蛋白可与多聚磷酸结合,发挥各种生物学功能.本文将简要介绍多聚磷酸相关蛋白的结构与主要生物学功能,以阐述多聚磷酸参与的细胞内生化过程.     

强化生物除磷系统的功能微生物研究进展

人类活动过程中排放的磷是导致水体富营养化的重要原因之一,因此,采取强化生物除磷(Enhanced biological phosphorus removal,EBPR)技术去除污水中磷,减轻对环境不利影响。由于具有经济、可持续的优点,EBPR系统在污水除磷中得到广泛应用,而体系中微生物群落组成合理、功能完整是EBPR系统高效稳定运行的关键所在。为了深入了解EBPR系统除磷机理和实现高效稳定运行,对系统中微生物群落结构和主要功能微生物进行了大量研究。EBPR系统中除了具有聚磷能力的聚磷菌(Polyphosphate-accumulating organisms,PAOs)外,还包括没有聚磷能力的非聚磷菌(non-PAOs),主要为聚糖菌(Glycogen-accumulating organisms,GAOs)和一些辅助细菌等。目前,发现与聚磷相关的功能微生物种类越来越多,研究最多的PAOs和GAOs分别为Accumulibacter和Defluviicoccus。PAOs和GAOs在不同的环境条件下存在竞争或合作关系,但是PAOs在特定条件下是否能够表现出GAOs的代谢特性这一问题还存在争论。除传统碳源、p H和温度等因素影响生物除磷外,外源污染物(如抗生素和重金属)对EBPR系统中功能微生物也产生影响。为了获得高效PAOs,传统分离方法、蓝白斑筛选法和人工构建工程菌的方法先后得到应用。现代分子生物学技术的发展为EBPR系统中功能微生物研究提供了先进可靠的技术手段,通过高效聚磷菌的构建实现高效除磷是未来提高实际污水中生物除磷效率的一个重要发展方向。     

聚磷生物膜法磷回收研究进展

从污水中回收磷已经成为当前污水全面资源化研究领域的重要研究方向之一,聚磷生物膜法磷回收工艺因其在节省碳源、简化回收流程、提高回收效果、污泥减量化等方面的潜力,成为该领域的重要技术手段和研究热点。本文着重介绍了聚磷生物膜法磷回收的基本原理、功能微生物及其代谢特征;详细梳理了生物膜法磷回收工艺不同模式的回收思路,综合分析了不同工艺模式的技术特征和磷回收效果。此外,对聚磷生物膜系统微生物及其代谢的研究方法与技术进行了整理。文章在综合分析聚磷生物膜法磷回收研究现状的基础上,对该技术的应用前景进行了宏观展望,以期为开展聚磷生物膜法磷回收的基础研究和技术开发提供支撑。     

一株高效聚磷解淀粉芽孢杆菌的分离鉴定及其除磷条件优化

[背景]磷是引起水体富营养化的主要营养物质.生物除磷具有成本低、效率高且适用范围广等特点,成为近年来水处理研究领域的热点.目前虽然有一些聚磷菌被筛选获得,但它们的除磷效率不高,其除磷条件尚需优化.聚磷菌的固定化及回收利用也亟待研究.[目的]分离筛选并鉴定高效聚磷菌株,优化除磷环境条件,研究吸附材料对所筛菌株除磷的影响,...     

高效嗜热聚磷菌的构建

以一株能聚磷但不耐热的芽孢杆菌和一株嗜热但不聚磷的嗜热菌为亲本,PEG诱导原生质体融合,筛选出2株既耐高温又能聚磷的融合菌株。对2株融合子进行传代培养,同时对未传代培养的融合子和传代培养后的融合子进行聚磷及耐热性测定,2株菌株的聚磷能力均比亲株高,其对无机磷的聚积能力分别为27.8%和5.8%,对有机磷的聚积能力分别为39.2%和56.1%,,耐热温度达55℃。该稳定的融合子具有嗜热和聚磷的双重遗传标记,可以作为研究基因重组的材料及高温生物除磷的生产菌株,对研究聚磷菌的聚磷机理和嗜热菌耐热机理,解决夏季污水处理曝气池发泡过多现象提供一定的依据。     

亚硝酸盐对污水生物除磷影响的研究进展

亚硝酸盐作为生物硝化和反硝化的中间产物, 存在于污水生物脱氮除磷系统中。对于生物强化除磷工艺亚硝酸盐既是电子受体用于反硝化除磷, 同时又是抑制剂影响生物除磷过程。本文综述了聚磷菌在厌氧、好氧和缺氧环境中的代谢机理, 在此基础上分别从好氧除磷和反硝化除磷两方面介绍了亚硝酸盐对污水生物除磷影响的研究, 同时概述了亚硝酸盐对生物除磷的抑制机理, 并对该领域的研究提出了个人见解。     

福建省3座水库库心沉积物聚磷菌的群落特征

聚磷菌在水库沉积物磷的代谢循环中起着重要作用.采用T-RFLP技术和克隆测序,以多聚磷酸盐激酶基因(ppk1)作为标记基因,研究对比了福建省3座富营养化水库(九龙江西陂水库、平潭三十六脚湖水库和三明东牙溪水库)库心沉积物中聚磷菌的多样性及群落结构.结果表明:3座水库库心沉积物聚磷菌的多样性存在一定差异,但不显著,以三十六脚湖库心样品中聚磷菌多样性最高(Shannon指数H=2.89±0.03,Simpson指数D=0.06±0.01).克隆测序和比对结果分析表明,三十六脚湖水库沉积物样品中聚磷菌种类最多,结构最复杂,这与T-RFLP的结果一致.3座水库库心沉积物样品中聚磷菌优势菌属差异较为明显,但均以变形菌门、放线菌门和酸杆菌门为主,这3个门在3座水库库心沉积物样品中所占比重分别达74.5%、85.0%和75.0%.其中,变形菌门中的厌氧粘细菌属、酸杆菌门中的Solibacter属在3座水库库心沉积物样品中均为优势菌属.沉积物中Fe/Al-P与聚磷菌多样性指数的相关性最显著.Fe/Al-P对聚磷菌群落结构影响最为显著,3个水库共有优势菌属厌氧粘细菌与各形态磷呈正相关,且与OP、Ca-P等难溶解性磷呈显著相关,Solibacter与各形态磷均呈负相关.表明聚磷菌与富营养化水库沉积物磷代谢循环密切相关.     

藻细胞的聚磷作用及其成矿意义

研究了现代蓝藻盐泽螺旋藻（Ｓｐｉｒｕｌｉｎａ ｓｕｂｓａｌｓａ）在富磷环境中细胞的聚磷作用及其与磷块岩形成的关系。实验证明,在富磷环境中盐泽螺旋藻细胞从环境中吸收磷的速度快,并主要用以合成多聚磷酸盐。模拟成矿试验证明,从藻细胞中提取出来的多聚磷酸盐在Ｃａ２＋ －ＨＰＯ２－４ －ＨＣＯ－３ －Ｆ－ －Ｈ２Ｏ 体系中,在常温常压及偏碱性条件下形成的主要成分为碳氟磷灰石、方解石及非晶质磷酸盐的矿物,为藻类成矿提供了新的极有意义的证据     

Polyphosphate in bone

Human bone-forming osteoblasts are an excellent model to investigate the multiple functions of inorganic polyphosphates (polyP) for the following reasons: 1) they contain relatively high amounts of polyP and polyP-dependent enzymes; 2) they allow the study of both general and specific functions of these polymers, and 3) medically relevant results can be expected from these studies.     

Inorganic Polyphosphate and Cancer

This review presents data on the relationship between inorganic polyphosphate metabolism and carcinogenesis including participation of polyphosphates in the regulation of activity of mTOR and other proteins involved in carcinogenesis, the role of h-prune protein (human polyphosphatase) in cell migration and metastasis formation, the prospects for using polyphosphates and inhibitors of polyphosphate metabolism enzymes as agents for controlling cell proliferation and migration.     

Polyphosphate synthesis in yeast

Polyphosphate synthesis was studied in phosphate-starved cells of Saccharomyces cerevisiae and Kluyveromyces marxianus. Incubation of these yeasts for a short time with phosphate and either glucose or ethanol resulted in the formation of polyphosphate with a short chain length. With increasing incubation times, polyphosphates with longer chain lengths were formed. Polyphosphates were synthesized faster during incubation with glucose than with ethanol. Antimycin did not affect the glucose-induced polyphosphate synthesis in either yeast. Using ethanol as an energy source, antimycin A treatment blocked both polyphosphate synthesis and accumulation of orthophosphate in the yeast S. cerevisiae. However, in K. marxianus, polyphosphate synthesis and orthophosphate accumulation proceeded normally in antimycin-treated cells, suggesting that endogenous reserves were used as energy source. This was confirmed in experiments, conducted in the absence of an exogenous energy source.     

Methods for Detection and Quantification of Polyphosphate and Polyphosphate Accumulating Microorganisms in Aquatic Sediments

It has been speculated that the microbial P pool is highly variable in the uppermost layer of various aquatic sediments, especially when an excessive P accumulation in form of polyphosphate (Poly‐P) occurs. Poly‐P storage is a universal feature of many different organisms and has been technically optimised in wastewater treatment plants (WWTP) with enhanced biological phosphorus removal (EBPR). In the recent past, new insights into mechanisms of P elimination in WWTP almost exclusively depended on the development and application of novel methods like ³¹P‐NMR spectroscopy and molecular methods for identifying Poly‐P accumulating microorganisms (PAO). The aim of the present review is to compile current methods potentially available for detection and quantification of Poly‐P in sediments and to complement it with yet unpublished results to validate their application in natural sediments. The most powerful tool for reliable Poly‐P quantification in sediments is the liquid ³¹P‐NMR technique which has been successfully used for Poly‐P measurements in a variety of aquatic sediments. But the microorganisms as well as mechanisms involved in Poly‐P storage and cycling are largely unknown. Therefore, we also intend to stimulate future studies focusing on these encouraging topics in sediment research via the implementation of novel methods. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Polyphosphate Is a Primordial Chaperone

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Inorganic Polyphosphate Modulates TRPM8 Channels

Polyphosphate (polyP) is an inorganic polymer built of tens to hundreds of phosphates, linked by high-energy phosphoanhydride bonds. PolyP forms complexes and modulates activities of many proteins including ion channels. Here we investigated the role of polyP in the function of the transient receptor potential melastatin 8 (TRPM8) channel. Using whole-cell patch-clamp and fluorescent calcium measurements we demonstrate that enzymatic breakdown of polyP by exopolyphosphatase (scPPX1) inhibits channel activity in human embryonic kidney and F-11 neuronal cells expressing TRPM8. We demonstrate that the TRPM8 channel protein is associated with polyP. Furthermore, addition of scPPX1 altered the voltage-dependence and blocked the activity of the purified TRPM8 channels reconstituted into planar lipid bilayers, where the activity of the channel was initiated by cold and menthol in the presence of phosphatidylinositol 4,5-biphosphate (PtdIns(4,5)P₂). The biochemical analysis of the TRPM8 protein also uncovered the presence of poly-(R)-3-hydroxybutyrate (PHB), which is frequently associated with polyP. We conclude that the TRPM8 protein forms a stable complex with polyP and its presence is essential for normal channel activity.     

Polyphosphate and stress response in mycobacteria

Polyphosphate (poly P) is present in every living cell. Long considered a 'molecular fossil', its role in cell physiology has been neglected. However, in the last few years it has become clear that poly P plays a role in multiple physiological functions, the best characterized of which is rpoS and recA induction during the Escherichia coli stringent response. Sureka et al. in this issue of Molecular Microbiology investigate the role of poly P in mycobacterial stress response and describe its participation in a novel regulatory pathway involving the two-component system MprAB, the alternative sigma factor sigma(E) and Rel, the enzyme responsible for (p)ppGpp metabolism in mycobacteria.