This opinion piece proposes that the environmental crises of our time arise from a failure to recognise the vital role of microbes in sustaining life on Earth, where ecosystems have been shaped for billions of years by microbial processes, including oxygen production, nutrient cycling and climate regulation. Yet the idea that microbes can 'teach' us how to navigate complexity, adapt across scales, and sustain planetary systems is still marginalised in science, policy, and education. A paradigm shift is proposed: microbes must be reframed as active collaborators in solving global challenges. This perspective is grounded in microbial ecology, Indigenous knowledge, and ethical philosophy, advocating for 'learning' through and with microbial life. To institutionalise this transition, policy and educational reforms are urged, centring microbial literacy as a foundation for ecological understanding. By integrating microbial agency into human knowledge systems, societal actions could be realigned with the biochemical and evolutionary logics that have sustained life for millennia. Ultimately, a deeper engagement with microbial knowledge is called for-one that informs a more sustainable future.

Microbial biotechnologies could affect health through multiple pathways, including impacts on food, nutrition, and the physical, economic, and social environment. Health in All Policies is an approach to ensure that plans and policies in all sectors maximise health gains and minimise any health risks. This approach often uses health impact assessment as a structured process to identify and assess positive and negative health impacts and make recommendations to improve these. There are very few examples where HIA has been applied to the implementation of microbial biotechnologies. As more biotechnologies are developed and implemented, more routine use of HIA could help to avoid adverse effects and realise their potential to improve health and reduce health inequalities. This will require greater awareness and understanding of the breadth of links to health, research to build the evidence base for these links, and governance mechanisms to oversee the development and implementation of microbial biotechnologies that prioritise health, equity and sustainability.

ABSTRACTN‐Acyl‐D‐amino acid deacylases (EC 3.5.1.81, also known as D‐acylases) have been studied for decades for their utility in the kinetic resolution of N‐acetyl‐D,L‐amino acids (NAAs) due to a marked stereospecificity. In conjunction with an N‐succinyl‐amino acid racemase (NSAR), they impulse the dynamic kinetic resolution (DKR) of different NAAs until the corresponding enantiomerically pure D‐amino acids. Besides the clear interest in this enzyme cascade, the application of D‐acylase/NSAR tandems has been only briefly described outside the industrial field. In this work, we revisit D‐acylases for the DKR of NAAs, reporting the characterisation of two new recombinant D‐acylases belonging to Bordetella petrii and Klebsiella pneumoniae. The enzymes were successfully coupled with the recombinant NSAR from Geobacillus stearothermophilus for the biosynthesis of D‐methionine or D‐aminobutyric acid. We also carried out the structural characterisation of the D‐acylase from Klebsiella pneumoniae (KleDacyl), providing the second experimental 3‐D structure of a member of this family of enzymes. The structural model shows a highly dynamic character of this amidohydrolase superfamily member, supplying a snapshot of an open conformation of the enzyme most likely preceding substrate entrance into the catalytic cleft. Our results confirm for the first time the importance of an α/β mobile domain in the substrate specificity of D‐acylases (region 282–341 in KleDacyl), opening up new strategies for structural‐based protein engineering strategies.

Efficient and economical purification methods are crucial for the commercial production of recombinant proteins with biomedical applications. In this study, we developed an affinity chromatography system that leverages the polysaccharide-binding properties of galectin-1 (GAL1) as a protein tag. The known GAL1-binding material, chitin, was used as the purification matrix. Melittin (MELT), a bee venom peptide known for its antimicrobial and anti-inflammatory properties with commercial potential, was chosen to validate this system. The GAL1-MELT fusion protein was expressed in Escherichia coli (E. coli) and successfully purified using a chitin-based matrix with sodium dodecyl sulfate (SDS) as a removable eluant. This method demonstrated higher purification efficiency compared to the His-tag/Ni-NTA approach, indicating that the GAL1/chitin system could serve as a superior alternative. The GAL1-MELT fusion protein retained strong antibacterial and anti-inflammatory activities, as well as collagen content modulation effects, confirming that MELT maintained its bioactivity. Apart from that, the GAL1-DsRed fusion protein was used as an additional protein target to evaluate the efficiency of the chitin-based column. Notably, all experiments were conducted without tag cleavage, showing that enzyme treatments for MELT isolation were unnecessary. This study highlights the potential of GAL1-polysaccharide interactions as a cost-effective and highly efficient alternative method for recombinant protein purification.

Psilocybin, a tryptamine-derived alkaloid, has been granted Breakthrough Therapy designation by the U.S. FDA for treatment-resistant depression, underscoring its clinical importance. Therefore, sustainable and economic production is urgently needed. Manufacturing of psilocybin in Escherichia coli has drawn great attention. However, due to the low expression and activity of the eukaryotic cytochrome P450 enzyme PsiH in the psilocybin biosynthetic pathway, de novo synthesis of psilocybin in prokaryotic cells has been hampered. To overcome this dilemma, we herein demonstrated de novo synthesis of psilocybin in E. coli by constructing PsiH variants with N-terminal domain modifications and expressing the entire biosynthetic pathway at a concordantly low temperature. Improving the supply of precursor and engineering the P450 electron transfer chain resulted in a 33-fold increase in the titre of norbaeocystin (105.3 mg/L), a key intermediate of psilocybin biosynthesis, and a 17-fold increase in the titre of psilocybin (14 mg/L). Further enhancement of psilocybin production was achieved by converting norbaeocystin to psilocybin by overexpressing an extra copy of the methyltransferase gene psiM. Finally, 79.4 mg/L of psilocybin was produced by optimising flask fermentation conditions, a 100-fold improvement over the starting strain. Our work demonstrates the successful fungal P450 engineering to improve the catalytic activity in E. coli and will advance the sustainable production of the important antidepressant psilocybin in prokaryotic microbial cells.

An upsurge of monkeypox disease (mpox) cases with clade I virus in Central Africa led WHO to declare a Public Health Emergency of International Concern for a second time shortly after the worldwide clade II mpox epidemic in 2022/3 among homosexual men. In the Democratic Republic of Congo (DRC), the annual incidence of clade I mpox, transmitted mostly from animal sources to children, increased 20-fold between 1980 and 2007; 60,000 mpox cases occurred between 2010 and 2023. The incidence again doubled between 2023 and 2024, showing a case fatality rate of 3.3%. A new clade Ib virus was detected in 2024 in eastern DRC where mostly adults were infected by heterosexual contact. Ib was recently introduced and showed a mutation spectrum of human-to-human transmission. Asymptomatic mpox infections, the release of infectious virus before symptom onset in a subgroup of cases, and superspreaders complicate containment measures during the 2022 epidemic. Isolation of cases until two consecutive negative PCR tests was recommended but necessitates cheap and rapid diagnostic tests which are in development. Sexual behavioural changes during the 2022 epidemic have contributed more to the curbing of the epidemic than vaccination. The smallpox vaccine Dryvax protected children exposed to clade I mpox in DRC in the 1980s. The attenuated third-generation smallpox Modified Vaccinia Ankara (MVA) vaccines and derivatives showed robust protection against clade IIb mpox during the 2022/3 epidemic in various study formats. Vaccine efficacy exceeding 75% was reported after two doses. mRNA in lipid-nanoparticle encoding surface proteins from extracellular enveloped and intracellular mature virions of monkeypox virus (MPXV) induced humoral and cellular immune responses that protected macaques against mpox disease with clade I and II viruses better than MVA. Only mixtures of monoclonal antibodies protected mice from mpox. The antiviral tecovirimat showed no efficacy in two clinical trials against clade I and II mpox.

In systems biocatalysis, combining pathway enzymes invitro allows for the conversion of basic substrates into more complex, valuable chemicals. However, invitro enzyme cascades are not yet economically viable for large-scale bio-based chemical production. Enhancing pathway efficiency through enzyme colocalization on synthetic protein scaffolds is a proposed solution, though still debated. We constructed a synthetic protein scaffold that colocalises the first three glycolytic enzymes using cohesin-dockerin interactions. Initially, we converted wild-type enzymes to the docking enzyme mode and evaluated their activity. Next, we demonstrate how the colocalisation of the three docking enzymes on distinct scaffolds enhances the enzyme cascade's production. Starting from glucose, the multi-enzyme complexes produced fructose-1,6-bisphosphate, confirming the activity of each enzyme. PfkA, which converts fructose-6-phosphate and ATP to fructose-1,6-bisphosphate and ADP, was identified as the rate-limiting enzyme. We demonstrated that scaffolding proximity effects lead to higher product output than free docking enzymes, particularly at lower enzyme densities. Further research is needed to determine the relevance of enzyme colocalisation under industrial production settings. In addition, optimising an enzyme cascade demands a thorough understanding of reaction mechanisms and kinetics. The VersaTile method streamlines optimisation studies of modular proteins and complexes, enabling analysis of a broader design space by bypassing technical preparatory hurdles.

The COVID-19 pandemic has caused unparalleled impacts on global social dynamics, healthcare systems and economies, highlighting the urgent need for effective interventions to address current challenges and future pandemic preparedness. This study introduces a novel virus neutralisation platform based on SimCells (~1 μm) and mini-SimCells (100-200 nm), which are chromosome-free and non-replicating bacteria from an LPS-free Escherichia coli strain (ClearColi). SimCells and mini-SimCells were engineered to display nanobodies on their surface, specifically targeting the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein - a critical immunogenic fragment essential for viral entry into host cells. It was demonstrated that nanobody-expressing SimCells achieved over 90% blocking efficiency against synthesised RBD from both the original Wuhan and the B.1.351 (Beta) variant using competitive enzyme-linked immunosorbent assay (ELISA) assay. More importantly, live virus neutralisation assays demonstrated that NB6 nanobody-presenting mini-SimCells effectively neutralised the live SARS-CoV-2 Victoria variant with an IC50 of 2.95 × 109 ± 1.40 × 108 mini-SimCells/mL. Similarly, VE nanobody-presenting mini-SimCells effectively neutralised the B.1.351 (Beta) variant of the SARS-CoV-2 virus with an IC50 of 5.68 × 109 ± 9.94 × 108 mini-SimCells/mL. The mini-SimCells successfully protected Vero cells, a cell line derived from the kidney of an African green monkey, from infection by the live virus of SARS-CoV-2 and its variants. These results suggest that SimCell-based neutralisation offers a promising strategy for the prevention and treatment of SARS-CoV-2, and potentially other viral infections.