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The demand for lithium production has seen a significant rise, with the growing electric vehicle and stationary battery markets requiring further development of sustainable and scalable extraction met...

Valores excessivos pagos por cirurgias da pele estão a ser apurados pelo hospital, que também recebeu do Estado pelos atos cirúrgicos feitos aos fins de semana. Depois será a ACSS a determinar se o Hospital tem dinheiro a devolver. Decisão será conhecida em dezembro

The development of hydrogenases capable of operating under oxygenic photosynthetic conditions remains a key challenge for sustainable biohydrogen production. In this study, we developed a series of chimeric NAD+-reducing [NiFe]-hydrogenases (SH) combining structural elements from the O2-tolerant SH of Cupriavidus necator (CnSH) and the ferredoxin-interacting SH of Synechocystis sp. PCC6803 (SynSH). By engineering chimeric HoxU and HoxF subunits, we developed constructs─MixSH, Ch-HoxEFSyn+UCn, and Ch-HoxUswapCTD─that successfully couple the CnHoxYH hydrogenase module to the SynHoxEFU reductase module while retaining O2 tolerance and enhancing interaction with reduced ferredoxin. The lithoautotrophic growth of C. necator confirmed the tolerance of these variants to O2, while activity assays in Synechocystis demonstrated partial hydrogenase function, including H2 consumption and fermentative H2 production. Notably, Ch-HoxEFSyn+UCn retained ferredoxin interaction despite lacking the [4Fe4S]U4 cluster, showing [2Fe2S]F2 in HoxF as a functional ferredoxin-binding site. Moreover, we achieved the artificial integration of a [2Fe2S] cluster into CnHoxF and identified the CnHoxF N-terminal domain as structurally and functionally analogous to SynHoxE. Although electron transfer efficiency and activity in Synechocystis remained limited, this work validates the modular engineering of [NiFe]-hydrogenases, uniting O2-tolerance with ferredoxin interaction and offering a foundational step toward photosynthesis-coupled H2 production.

While Yarrowia lipolytica has gained prominence as a microbial chassis for biomanufacturing, its broader application faces two critical limitations: incomplete genetic annotation and insufficient char...