Proteomic Analysis of 2,4,6-Trinitrotoluene Degrading Yeast Yarrowia lipolytica

Abstract

2,4,6-trinitrotoluene (TNT) is a common component of many explosives. The overproduction and extensive usage of TNT significantly contaminates the environment. TNT accumulates in soils and aquatic ecosystems and can primarily be destroyed by microorganisms. Current work is devoted to investigation of Yarrowia lipolytica proteins responsible for TNT transformation through the pathway leading to protonated Meisenheimer complexes and nitrite release. Here, we identified a unique set of upregulated membrane and cytosolic proteins of Y. lipolytica, which biosynthesis increased during TNT transformation through TNT-monohydride-Meisenheimer complexes in the first step of TNT degradation, through TNT-dihydride-Meisenheimer complexes in the second step, and the aromatic ring denitration and degradation in the last step. We established that the production of oxidoreductases, namely, NADH flavin oxidoreductases and NAD(P)+-dependent aldehyde dehydrogenases, as well as transferases was enhanced at all stages of the TNT transformation by Y. lipolytica. The up-regulation of several stress response proteins (superoxide dismutase, catalase, glutathione peroxidase, and glutathione S-transferase) was also detected. The involvement of intracellular nitric oxide dioxygenase in NO formation during nitrite oxidation was shown. Our results present at the first time the full proteome analysis of Y. lipolytica yeast, destructor of TNT.

Degradation of 2,4,6-Trinitrotoluene (TNT): Involvement of Protocatechuate 3,4-Dioxygenase (P34O) in Buttiauxella sp. S19-1

“…However, the conventional methods for remediation of aromatic pollutants, such as chemical oxidation, hydrolysis, and incineration, are costly and complex [16]. Indeed, there is strong evidence supporting TNT-degrading abilities of fungi [17][18][19][20] and plants [21][22][23]. Furthermore, previous studies showed that several bacteria, such as Pseudomonas [1], Bacillus [24], Enterobacter [25], and Rhodococcus [26], were capable of degrading TNT.…”

Review of Explosive Contamination and Bioremediation: Insights from Microbial and Bio-Omic Approaches

“…Proteomics analyses can vary, from studying a single organism or strain to conducting differential zone analyses. For example, proteomic research in Yarrowia lipolytica revealed the expression of proteins related to membrane-bound oxidoreductases, which play a pivotal role in triggering the transformation of TNT [ 55 ]. This information sheds light on the biological degradation of explosives in plants and fungi and highlights the complex changes in proteins during pollutant bioremediation, even so one limitation of proteomics analysis is its focus on individual organisms or strains, and understanding the proteomics of an entire microbial community remains a challenge [ 55 , 56 ].…”

Synthetic biology tools for engineering Yarrowia lipolytica

“…A separate transcriptome analysis, carried out in tandem with proteomics methods, was used to explore amino acid catabolism ( Mansour et al, 2009 ; Morín et al, 2007 ). Additionally, proteome analyses were conducted in Y. lipolytica to characterize the proteins involved in the yeast-to-hypha transition ( Morín et al, 2007 ); the osmotic response to erythritol ( Yang et al, 2015 ); and the degradation of TNT ( Khilyas et al, 2017 ). Fluxomics has grown as a discipline thanks to 13C-based metabolic flux analysis.…”