Isolation and Characterization of Novel Hyperthermophilic Manganese (II)-oxidase from Purple Pool Yellowstone National Park (2021-03-12)

Abstract

Manganese (Mn) oxides are some of the strongest naturally occurring oxides in nature,playing an important role in geochemical cycling, and living systems. Several theories onthe role of Mn in microbial systems exist, but there is not enough supporting evidence toprovide a strong argument for these theories. Understanding the role of Mn in microbialsystems will allow for further investigations into the evolution of early life, defensivemechanisms, and electron transfer systems in microbes. Preliminary sequence data ofbiofilms and enrichments of biofilms collected from a Mn-depositing hot spring (102ºC) inYellow Stone National Park (YNP), have yielded findings that suggest a novelhyperthermophilic archaea may be responsible for the accumulation of Mn(III/IV) oxideswithin the hot spring. There are currently no known hyperthermophilic archaea that iscapable of oxidizing Mn(II), an investigation into the properties of the Mn oxidizingproperties will have wide impacts in fields of early life, bioremediation, and industry. Theinvestigation will involve biofilm studies, to study its role in Mn oxidation mechanisms, andalso in Mn oxide morphologies as a respect of Mn concentration and flow rate. Thesestudies would be useful in biomarker analysis in Mn containing geological features, mostnotably in YNP. A bioinformatics component of the investigation will involve aninvestigation of the genome and mapping of possible gene sequences responsible for Mnoxidation. The Mn oxidase genes will then be further contextualized by constructingmetabolic pathways of the genomes. The biochemical component will include the isolationand expression, and synthesis of the proteins from the sequenced identified for Mnoxidation. As a protein originating from hyperthermophilic archaea, it is hypothesized that itwill have retained activity in high temperatures. These hypothesis will be tested by runningactivity assays, in respect to product/intermediate formations. In addition, denaturationconditions will be tested to examine the durability of the protein structure. Characterizationof the novel Mn oxidation components of the thermophilic archaea will be impactful infields of environmental science, bioinorganic chemistry, and industry.