Identification of the Dinuclear Metallo-Cofactor and Elucidation of the Reaction Mechanism of the Aldehyde Decarbonylase from the Alkane-producing Cyanobacterium Nostoc Punctiforme

产烷烃点状发菜蓝藻双核金属辅因子的鉴定及醛脱羰酶反应机制的阐明

• 批准号: 1122079
• 负责人: Carsten Krebs
• 金额: $ 81.28万
• 依托单位: Pennsylvania State Univ University Park
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1122079&HistoricalAwards=false
• 关键词: Identification; Dinuclear; Metallo; Cofactor; Elucidation

This award in the molecular biosciences program supports work at the Pennsylvania State University to study the enzyme aldehyde decarbonylase (AD) from the alkane-producing cyanobacterium Nostoc punctiforme (Np). The reaction catalyzed by Np AD is the second of the two reactions in the biosynthesis of alkanes by cyanobacteria. Thus, research supported by this project aims at elucidating the detailed chemical steps of the key alkane-producing step in cyanobacteria. Because cyanobacteria are environmentally versatile, rapidly growing, genetically alterable, and (most importantly) photosynthetic, they are being actively pursued as a vehicle for development of renewable energy. These studies include the identification of the dinuclear metallocofactor required for activity of Np AD and determination of the reaction mechanism of Np AD by a combination of biochemical and biophysical approaches. The utilization of fatty acids suggests that the soluble Np AD has devised a novel strategy to obtain this water insoluble substrate. The mechanism of substrate acquisition by Np AD will also be evaluated to differentiate between a channeling mechanism and membrane targeting. This project uses a versatile set of experimental strategies to evaluate the biosynthesis of alkanes. The results from these proposed studies will provide valuable information on new chemical properties of an enzyme metal center. The broader impacts of this work are two-fold. First, the research area at the interface of biological and inorganic chemistry is very interdisciplinary and offers students and postdoctoral scholars to be trained in a wide variety of methods. A workshop will be offered by bioinorganic chemists from Penn State and other institutions to approximately 80 students and postdoctoral scholars as a unique training opportunity in many of the methods used in this research field. Second, the research is relevant for the development of renewable energy, because the use of cyanobacteria for alkane production is a promising strategy for biofuel production. This process is particularly environmentally friendly, because the process is (i) carbon neutral (all carbon found in the alkane product derives from carbon dioxide fixed by the cyanobacteria) and (ii) driven by solar energy (due to the photosynthetic nature of cyanobacteria). This project will aid in the rational design of genetically engineered cyanobacteria for enhanced biofuel production.

该分子生物科学项目奖项支持宾夕法尼亚州立大学研究来自产烷烃蓝细菌点状发菜 (Np) 的醛脱羰酶 (AD) 的工作。 Np AD 催化的反应是蓝细菌生物合成烷烃的两个反应中的第二个反应。因此，该项目支持的研究旨在阐明蓝藻中关键烷烃生产步骤的详细化学步骤。由于蓝藻具有环境多功能性、生长迅速、基因可变以及（最重要的是）光合作用，因此人们正在积极将其作为开发可再生能源的工具。这些研究包括鉴定 Np AD 活性所需的双核金属辅因子，以及通过生物化学和生物物理相结合的方法确定 Np AD 的反应机制。脂肪酸的利用表明可溶性 Np AD 设计了一种新的策略来获得这种水不溶性底物。还将评估 Np AD 获取底物的机制，以区分通道机制和膜靶向。该项目使用一套多功能的实验策略来评估烷烃的生物合成。 这些拟议研究的结果将为酶金属中心的新化学性质提供有价值的信息。这项工作的更广泛影响有两个方面。首先，生物化学和无机化学交叉领域的研究领域具有很强的跨学科性，为学生和博士后学者提供了多种方法的培训。来自宾夕法尼亚州立大学和其他机构的生物无机化学家将为大约 80 名学生和博士后学者举办研讨会，作为该研究领域使用的许多方法的独特培训机会。其次，该研究与可再生能源的发展相关，因为利用蓝细菌生产烷烃是生物燃料生产的一种有前景的策略。该过程特别环保，因为该过程是（i）碳中性（烷烃产物中发现的所有碳均来自蓝藻固定的二氧化碳）和（ii）由太阳能驱动（由于蓝藻的光合作用性质）。该项目将有助于合理设计基因工程蓝细菌，以提高生物燃料的生产。