DOI: 10.5281/zenodo.15839119
title: Dataset for "Engineering O2-Tolerant Chimeric Hydrogenases Optimized for Ferredoxin Coupling in Synechocystis sp. PCC 6803"

description: The uploaded files contain data on the hydrogenase activities of Synechocystis sp. PCC 6803 strains expressing chimeric NAD+-reducing [NiFe]-hydrogenases (SH), which combine structural elements from the O2-tolerant SH of Cupriavidus necator and the ferredoxin-interacting SH of Synechocystis sp. PCC 6803. The data include hydrogen consumption / production activities and / or the measured H2 values and corresponding calculations. Graphical illustrations of the results are presented in the publication “Engineering O2-tolerant chimeric hydrogenases optimized for ferredoxin coupling in Synechocystis sp. PCC 6803“.
description: “H2 consumption_GC_Synechocystis chimeras_genome integration.xlsx”: 

Data on H2 consumption measurements of Synechocystis sp. PCC 6803 strains bearing chimeric hydrogenases integrated into the genome obtained from gas chromatography (GC) analysis. 5ml Liquid cultures of Syn_CnSH, Syn_HoxYHCn, Syn_MixSH, and Syn_CnSHHoxEF-fusion were flushed with 20% H2, 10% CO2, and 70% N2 in sealed 10 ml glass vials and incubated in the presence of light. GC measurements were performed immediately after flushing (0h) as well as after 6h, 18h, 24h, 48h and 72h of incubation. The file includes (1) hydrogenase activities (U/gCDW), (2) calculations from the obtained GC data (H2 %) as well as (3+4) background information on the applied method and strains.

The data correspond to Figure S15A of the publication “Engineering O2-tolerant chimeric hydrogenases optimized for ferredoxin coupling in Synechocystis sp. PCC 6803“. 

 

“H2 consumption_GC with DCMU_Synechocystis chimeras_genome integration.xlsx”: 

Data on H2 consumption measurements of Synechocystis sp. PCC 6803 strains bearing chimeric hydrogenases integrated into the genome and supplemented with DCMU obtained from gas chromatography (GC) analysis. 5ml Liquid cultures of Syn_CnSH, Syn_HoxYHCn, Syn_MixSH, and Syn_CnSHHoxEF-fusion supplemented with 20 µM DCMU were flushed with 20% H2, 10% CO2, and 70% N2 in sealed 10 ml glass vials and incubated in the presence of light. GC measurements were performed immediately after flushing (0h) as well as after 6h, 18h, 24h, 48h and 72h of incubation. The file includes (1) hydrogenase activities (U/gCDW), (2) calculations from the obtained GC data (H2 %) as well as (3+4) background information on the applied method and strains.

The data correspond to Figure S15B of the publication “Engineering O2-tolerant chimeric hydrogenases optimized for ferredoxin coupling in Synechocystis sp. PCC 6803“. 

 

„H2 consumption_GC_Synechocystis chimeras_replicative plasmid.xlsx“:

Data on H2 consumption measurements of Synechocystis sp. PCC 6803 strains bearing chimeric hydrogenases on replicative plasmids obtained from gas chromatography (GC) analysis. 5ml Liquid cultures of Syn_pCnSH, Syn_pMixSH, Syn_pCnSHHoxEF-fusion, Syn_pCnSHΔNTD-HoxF+HoxE, Syn_pCnSH+HoxE, Syn_pCh-HoxEFSyn+UCn and Syn_pCh-HoxUswapCTD induced with 2.5 µM Ni2+ for 24h were flushed with 20% H2, 10% CO2, and 70% N2 in sealed 20 ml glass vials and incubated in the presence of light. GC measurements were performed immediately after flushing (0h)  as well as after 2h, 4h, 6h and 8h of incubation. The file includes (1) hydrogenase activities (U/gCDW), (2) GC data (H2 %) as well as (3+4) background information on the applied method and strains.

The data correspond to Figure 5C and Table S4 of the publication “Engineering O2-tolerant chimeric hydrogenases optimized for ferredoxin coupling in Synechocystis sp. PCC 6803“.

 

„H2 production_Clark electrode_Synechocystis chimeras_genome integration.xlsx“:

Data on H2 production measurements of Syn_CnSH, Syn_Δhox’, Syn_MixSH, and Syn_CnSHHoxEF-fusion cultures using a Clark-type electrode and dithionite-reduced methyl viologen (MV) as an artificial electron donor. The file includes (1) hydrogenase activities (U/gCDW), (2) calculations as well as (3+4) background information on the applied method and strains.

The data correspond to Figure 5A of the publication “Engineering O2-tolerant chimeric hydrogenases optimized for ferredoxin coupling in Synechocystis sp. PCC 6803“.

 

„H2 production_Unisense electrode_Synechocystis chimeras_genome integration.xlsx“:

Data on H2 production measurements of Syn_CnSH, Syn_Δhox’, Syn_MixSH, and Syn_CnSHHoxEF-fusion cultures under fermenttive and photosynthetic conditions using a Unisense electrode. The file includes (1) H2 % values well as (2+3) background information on the applied method and strains.

The data correspond to Figure 5B and Figure S16 of the publication “Engineering O2-tolerant chimeric hydrogenases optimized for ferredoxin coupling in Synechocystis sp. PCC 6803“.