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Proteomic analysis of natural photoheterotrophic mixed consortium for biohydrogen production under nongrowing conditions

ABSTRACT: A biohydrogen and polyhydroxyalkanoates(PHA)-producing natural photoheterotrophic mixed culture composed mainly by Rhodopseudomonas palustris and Clostridium sp was studied by a proteomic analysis under non-growth conditions (nitrogen-absence and organic acids). Proteins in C. pasteurianum were upregulated, particularly those related to stress response. In contrast, C. pasteurianum in the consortium did not present such proteins, showing the advantage of being part of it. Both cultures showed proteins involved in organic acid metabolism and biohydrogen production, such as lactate dehydrogenase, ferredoxins, and hydrogenases. Proteomes of R. palustris as single culture and in consortium showed that organic acids were redirect into central carbon pathways to generate reduced equivalents for biohydrogen production. Light-harvesting proteins and fatty acid metabolism linked to PHA accumulation were also upregulated. This study provides insights into how the proteomes of individual organisms and their consortium counterparts adapt to non-growth conditions, shedding light on how microbial interactions influence protein expression.

INSTRUMENT(S): Orbitrap Exploris 480

ORGANISM(S): Clostridium Pasteurianum Dsm 525 = Atcc 6013 Rhodopseudomonas Palustris

TISSUE(S): Cell Culture

SUBMITTER: Timothy McCubbin

LAB HEAD: Elvia Ines Garcia Pena

PROVIDER: PXD057021 | Pride | 2025-02-06

REPOSITORIES: pride

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Project description:Peripheral light harvesting (LH) antenna complexes have been studied extensively in the purple nonsulfur bacterium Rhodopseudomonas palustris because it produces different types of LH complexes under high light intensities (LH2 complex) and low light intensities (LH3 and LH4 complex). The ability of R. palustris to alter its peripheral LH complexes in response to changes in light intensity is attributed to the multiple operons that encode the a and b peptides that make up these complexes, whose expression is affected by light intensity, light quality, and oxygen tension. However, low resolution structures, amino acid similarities between the complexes, and a lack of transcriptional analysis made it difficult to determine the LH complexes composition and functions under different light intensities. It was also unclear how much diversity of the R. palustris LH complexes exists in nature.Results: To gain insight into the composition of the LH complexes, their function under high light intensities and low light intensities, and their prevalence in the environment we undertook an integrative genomics approach using 15 closely related R. palustris strains isolated from the environment and 5 R. palustris ecotypes whose genomes have been sequenced. We sequenced the genomes for the 15 closely related strains and using RNA-seq carried out transcriptomic analysis on all 20 strains grown under high light intensity and low light intensity. We were able to determine that even closely related R. palustris strains had differences in their pucBA gene content and expression, even under the same growth conditions. We also found that the LH2 complex could compensate for the lack of an LH4 complex under LL intensities but not under extremely LL intensities. Conclusions: This is the first time an integrative genomics approach has been used to study light harvesting in the environment. The variation observed in LH gene composition and expression in environmental isolates of R. palustris likely reflects how these strains have adapted to specific light conditions in the environment. We have also shown that there is redundancy between some of the LH complexes under certain light intensities, which may partially explain why multiple operons encoding LH complexes have evolved and been maintained in R. palustris. Examing the variation observed in LH gene composition and expression in various environmental isolates

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Proteomic analysis of natural photoheterotrophic mixed consortium for biohydrogen production under nongrowing conditions

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