PETROX - Petroleum Alkane oxidation by Anaerobic Archaea

石油正构烷烃是原油中的主要化合物类别，其丰度决定了原油的流动性和可开采性。对于好氧和厌氧微生物来说，石油烷烃也是一种有吸引力的能源和碳源。这项建议旨在利用最近获得的独特的假丝酵母(ANME-1古菌的近亲)和假丝酵母Ca的培养来解决厌氧、石油烷烃氧化古菌的功能问题。Cerberiaceum(哈德氏菌深分枝属的一员)。这两种微生物都与脱硫杆菌作为硫酸盐还原伙伴一起生长。古菌用烷基辅酶M还原酶激活中到长链正构烷烃(C6-C20)，烷基辅酶M还原酶是甲基辅酶M还原酶的不同变体，这种酶已知可以产生或捕获甲烷。古生菌可能是地下石油转化的关键角色，具有耐人寻味的未知生物化学。在目前的PETROX项目中，我们将解决古生菌中石油烷烃降解的功能。围绕着一个为期31年半的博士项目，我们将(I)优化两种培养物的生长条件，并确定古生菌及其伙伴适应溶剂型底物的策略(Ii)使用天然结晶通过ACR解决石油烷烃激活的分子基础(Iii)在生理实验、稳定同位素探测实验和超微结构分析中评估古生菌与其细菌伙伴的相互作用。我们结合了我们在厌氧烃微生物学和生物化学方面的专业知识，并与可视化技术方面的专家合作。该项目将提供关于地下微生物组成员功能的基础知识，即激活长链烷烃和合养相互作用的酶机制。

Petroleum n-alkanes are a major compound class in crude oil, and their abundance determines the oils’ fluidity and exploitability. Petroleum alkanes are also an attractive energy and carbon source for aerobic and anaerobic microorganisms. This proposal aims at resolving the functioning of the anaerobic, petroleum-alkane oxidizing archaea using recently obtained, unique cultures of Candidatus Alkanophaga (a relative of the ANME-1 archaea) and Candidatus Ca. Cerberiarchaeum (a member of the deep-branching Hadarchaea). Both organisms grow with Thermodesulfobacteria as sulfate-reducing partners. The archaea activate mid- to long-chain n-alkanes (C6-C20) with alkyl coenzyme M reductases, divergent variants of methyl coenzyme M reductase, the enzyme known to generate or capture methane. The archaea are likely key players in subsurface oil transformations and have intriguing uncharted biochemistry. In the presented project PETROX, we will resolve the functioning of petroleum alkane degradation in archaea. Centralized around a 31/2-year PhD project, we will (I) optimize the growth conditions from both cultures and determine the strategies of the archaea and their partner to adapt to the solvent-like substrate (II) resolve the molecular basis of petroleum alkane activation via ACR using native crystallization (III) assess the interaction of the archaea with their bacterial partners in physiological experiments, stable isotope probing experiments and ultrastructural analyses. We combine our expertise in anaerobic hydrocarbon microbiology and biochemistry, and collaborate with experts in visualization techniques. This project will provide fundamental knowledge on the functioning of members from the subsurface microbiome, the enzyme mechanistic to activate long-chain alkanes and syntrophic interactions.