Dataset Information

Fixing N₂ into cyanophycin: continuous cultivation of Nostoc sp. PCC 7120.

ABSTRACT: Two diazotrophic cyanobacteria (Anabaena cylindrica PCC 7122 and Nostoc sp. PCC 7120) were cultivated to produce cyanophycin, a nitrogen reserve compound, under nitrogen fixing conditions. In preliminary continuous experiments, Nostoc sp. was shown to be more efficient, accumulating a higher amount of cyanophycin and showing a greater capability to fix atmospheric nitrogen in the biomass (67 mg_N d^-1 of fixed nitrogen per liter of culture). The operating conditions were then optimized to maximize the cyanophycin productivity: the effect of incident light intensity, residence time and nitrogen availability were investigated. Nitrogen availability and/or pH played a major role with respect to biomass production, whereas phosphorus limitation was the main variable to maximize cyanophycin accumulation. In this way, it was possible to achieve a stable and continuous production of cyanophycin (CGP) under diazotrophic conditions, obtaining a maximum cyanophycin productivity of 15 mg_CGP L^-1 d^-1. KEY POINTS: • Diazotrophic cyanobacteria produce stable amount of cyanophycin in continuous PBR. • Nostoc sp. proved to be more efficient in producing cyanophycin than Anabaena sp. • P deprivation is the major variable to increase cyanophycin productivity in continuous.

SUBMITTER: Trentin G

PROVIDER: S-EPMC9750909 | biostudies-literature | 2023 Jan

REPOSITORIES: biostudies-literature

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Fixing N2 into cyanophycin: continuous cultivation of Nostoc sp. PCC 7120.

Trentin Giulia G Piazza Francesca F Carletti Marta M Zorin Boris B Khozin-Goldberg Inna I Bertucco Alberto A Sforza Eleonora E

Applied microbiology and biotechnology 20221126 1

Two diazotrophic cyanobacteria (Anabaena cylindrica PCC 7122 and Nostoc sp. PCC 7120) were cultivated to produce cyanophycin, a nitrogen reserve compound, under nitrogen fixing conditions. In preliminary continuous experiments, Nostoc sp. was shown to be more efficient, accumulating a higher amount of cyanophycin and showing a greater capability to fix atmospheric nitrogen in the biomass (67 mgN d-1 of fixed nitrogen per liter of culture). The operating conditions were then ...[more]

PMID: 36435857

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Project description:BackgroundThe maturation of hydrogenases into active enzymes is a complex process and e.g. a correctly assembled active site requires the involvement of at least seven proteins, encoded by hypABCDEF and a hydrogenase specific protease, encoded either by hupW or hoxW. The N2-fixing cyanobacterium Nostoc sp. strain PCC 7120 may contain both an uptake and a bidirectional hydrogenase. The present study addresses the presence and expression of hyp-genes in Nostoc sp. strain PCC 7120.ResultsRT-PCRs demonstrated that the six hyp-genes together with one ORF may be transcribed as a single operon. Transcriptional start points (TSPs) were identified 280 bp upstream from hypF and 445 bp upstream of hypC, respectively, demonstrating the existence of several transcripts. In addition, five upstream ORFs located in between hupSL, encoding the small and large subunits of the uptake hydrogenase, and the hyp-operon, and two downstream ORFs from the hyp-genes were shown to be part of the same transcript unit. A third TSP was identified 45 bp upstream of asr0689, the first of five ORFs in this operon. The ORFs are annotated as encoding unknown proteins, with the exception of alr0692 which is identified as a NifU-like protein. Orthologues of the four ORFs asr0689-alr0692, with a highly conserved genomic arrangement positioned between hupSL, and the hyp genes are found in several other N2-fixing cyanobacteria, but are absent in non N2-fixing cyanobacteria with only the bidirectional hydrogenase. Short conserved sequences were found in six intergenic regions of the extended hyp-operon, appearing between 11 and 79 times in the genome.ConclusionThis study demonstrated that five ORFs upstream of the hyp-gene cluster are co-transcribed with the hyp-genes, and identified three TSPs in the extended hyp-gene cluster in Nostoc sp. strain PCC 7120. This may indicate a function related to the assembly of a functional uptake hydrogenase, hypothetically in the assembly of the small subunit of the enzyme.

Project description:BackgroundThe last step in the maturation process of the large subunit of [NiFe]-hydrogenases is a proteolytic cleavage of the C-terminal by a hydrogenase specific protease. Contrary to other accessory proteins these hydrogenase proteases are believed to be specific whereby one type of hydrogenases specific protease only cleaves one type of hydrogenase. In cyanobacteria this is achieved by the gene product of either hupW or hoxW, specific for the uptake or the bidirectional hydrogenase respectively. The filamentous cyanobacteria Nostoc punctiforme ATCC 29133 and Nostoc sp strain PCC 7120 may contain a single uptake hydrogenase or both an uptake and a bidirectional hydrogenase respectively.ResultsIn order to examine these proteases in cyanobacteria, transcriptional analyses were performed of hupW in Nostoc punctiforme ATCC 29133 and hupW and hoxW in Nostoc sp. strain PCC 7120. These studies revealed numerous transcriptional start points together with putative binding sites for NtcA (hupW) and LexA (hoxW). In order to investigate the diversity and specificity among hydrogeanse specific proteases we constructed a phylogenetic tree which revealed several subgroups that showed a striking resemblance to the subgroups previously described for [NiFe]-hydrogenases. Additionally the proteases specificity was also addressed by amino acid sequence analysis and protein-protein docking experiments with 3D-models derived from bioinformatic studies. These studies revealed a so called "HOXBOX"; an amino acid sequence specific for protease of Hox-type which might be involved in docking with the large subunit of the hydrogenase.ConclusionOur findings suggest that the hydrogenase specific proteases are under similar regulatory control as the hydrogenases they cleave. The result from the phylogenetic study also indicates that the hydrogenase and the protease have co-evolved since ancient time and suggests that at least one major horizontal gene transfer has occurred. This co-evolution could be the result of a close interaction between the protease and the large subunit of the [NiFe]-hydrogenases, a theory supported by protein-protein docking experiments performed with 3D-models. Finally we present data that may explain the specificity seen among hydrogenase specific proteases, the so called "HOXBOX"; an amino acid sequence specific for proteases of Hox-type. This opens the door for more detailed studies of the specificity found among hydrogenase specific proteases and the structural properties behind it.

Dataset Information

Fixing N₂ into cyanophycin: continuous cultivation of Nostoc sp. PCC 7120.

Publications

Fixing N<sub>2</sub> into cyanophycin: continuous cultivation of Nostoc sp. PCC 7120.

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