Biosynthesis and Monitoring of the Cyanobacterial Toxin Anatoxin-a(s)

蓝藻毒素 Anatoxin-a(s) 的生物合成和监测

基本信息

批准号：
10042382
负责人：
BRADLEY S MOORE
金额：
$ 19.69万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-13 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10042382
关键词：
Acetylcholinesterase Acute Address Aldehyde-Lyases Alkaloids Anabolism Anatoxins Animals Arginine Biochemical Bioremediations Chemicals Cholinesterase Inhibitors Clinical Communities Cyanobacterium Data Data Set Detection Development Diagnostic Dose Drug Metabolic Detoxication Economics Emergency Situation Environment Environmental Health Environmental Monitoring Enzymes Event Floods Florida Frequencies Fresh Water Gene Cluster Genes Genetic Screening Genomics Goals Health Human Industry Standard Knowledge Lead Lethal Dose 50 Metagenomics Monitor Mus Names Neurotoxins Ohio Organism Organophosphates Outcomes Research Paraoxon Pathway interactions Phosphoric Monoester Hydrolases Prevalence Privatization Production Reaction Recombinants Reporting Research Project Grants Safety Sarin Series Testing Time Torque Toxin Transaminases Transcript Tropanes Tube Water Water Supply Work anthropogenesis base chemical standard cyanobacterial toxin enzyme biosynthesis experience experimental study gene discovery harmful algal blooms metatranscriptomics neurotoxic public health relevance reconstitution

项目摘要

Project Summary / Abstract Freshwater harmful algal blooms of cyanobacteria or cyanoHABs are increasing in frequency and impact worldwide due to natural and anthropogenic flooding events. In the USA, all 50 states now regularly experience major cyanoHAB incidents, including Florida and Ohio that have declared states of emergency in recent years to address increased cyanotoxin loads in public waterways posing human health, environmental, and economic threats. While harmful cyanobacteria and their toxins are actively monitored, one of the most neurotoxic cyanotoxins, the organophosphate anatoxin-a(s), is not monitored. This water-soluble, UV-insensitive, and zwitterionic toxin is notoriously reactive, which has led to the ongoing challenges in its environmental monitoring. We recently discovered the genes encoding anatoxin-a(s) biosynthesis from the planktonic cyanobacterium Sphaerospermopsis torques-reginae ITEP-024. Through a series of genomic, chemical, and biochemical experiments, we have nearly reconstituted the entire biosynthetic pathway from arginine to anatoxin-a(s) with recombinant enzymes. Moreover, we identified nearly complete anatoxin-a(s) transcripts from the nearshore Western Basin of Lake Erie at Toledo, Ohio, suggesting that residents in Toledo as well as other communities across the USA may experience anatoxin-a(s) exposure without their knowledge and that of regional and national monitoring agencies. Our discovery sets the stage for this 2-year, R21 application to apply our biosynthetic expertise to fill the gap in knowledge about the prevalence, significance, and impact of this critical toxin in the environment. We propose four specific aims to address our broad goals. First, we plan to complete the functional assignment of all anatoxin-a(s) biosynthesis enzymes. Second, we will broadly analyze metagenomic and metatranscriptomic fresh water supplies for anatoxin-a(s) genes and to identify the producing cyanobacteria. Third, we aim to develop a rapid, PCR screen specific for environmental anatoxin- a(s) gene detection. And fourth, we will evaluate the native phosphatase AnsH and commercial phosphatases as anatoxin-a(s) degradation enzymes with potential bioremediation applications.

项目摘要 /摘要蓝细菌或氰基栖息的淡水有害藻华频率和影响正在增加由于自然和人为的洪水事件，全球范围内。在美国，所有50个州现在都经常体验近年来宣布紧急状态在内的重大蓝绿色事件，包括佛罗里达州和俄亥俄州解决构成人类健康，环境和经济状况的公共水道中增加的氰毒素负荷威胁。虽然有害的蓝细菌及其毒素被积极监测，但神经毒性之一是氰毒素，有机磷酸盐Anatoxin-A（S）未经监测。这种水溶性，不敏感的和奇异的毒素是反应性的，这导致了环境中的持续挑战监视。我们最近发现了浮游生物的编码Anatoxin-A（S）生物合成的基因 Sphaerospermopsis torques-Reginae itep-024。通过一系列基因组，化学和生化实验，我们几乎重构了整个生物合成途径从精氨酸到带有重组酶的Anatoxin-A（S）。此外，我们几乎确定了几乎完整的Anatoxin-A（S）转录本从俄亥俄州托莱多的伊利湖的近岸西部盆地，暗示托莱多和美国其他社区可能会在不知情的情况下经历Anatoxin-A的暴露，并且区域和国家监测机构。我们的发现为这项为期两年的R21申请奠定了基础应用我们的生物合成专业知识来填补有关患病率，意义和影响的知识的空白这种关键的毒素在环境中。我们提出了四个特定的目标，以解决我们的广泛目标。首先，我们计划完成所有Anatoxin-A（S）生物合成酶的功能分配。其次，我们将广泛分析Anatoxin-A（S）基因的元基因组和元文字淡水供应，并鉴定产生蓝细菌。第三，我们旨在开发针对环境解剖素的快速，PCR屏幕 - A（S）基因检测。第四，我们将评估天然磷酸酶ANSH和商业磷酸酶作为具有潜在生物修复应用的Anatoxin-A（S）降解酶。