Challenging plastic pollution with hydrocarbonoclastic lineages
The hydrocarbonoclastic lineages that have existed for millennia are responsible for the degradation of diverse aliphatic and aromatic compounds, regulating the ocean hydrocarbon cycles. Given the metabolic similarities in breaking down plastics and hydrocarbons, a thorough understanding and leveraging of these processes can provide biotechnologically based solutions to tackle global plastic pollution.
Laws and green incentives: guiding China's new biomass energy future
In this article, we focus on green incentives and laws guiding China's new biomass energy future. We offer proposals to reinforce green incentives and legal standards in this field.
Advancing biopharmaceutical manufacturing: economic and sustainability assessment of end-to-end continuous production of monoclonal antibodies
Monoclonal antibodies (mAbs) have become essential therapeutics for treating various diseases. The robust, cost-effective, and sustainable production of mAbs is crucial due to their growing clinical and commercial demand. Advances in bioprocessing, such as improved cell lines, perfusion bioreactors, multicolumn chromatography, and automation, can significantly increase productivity, making treatments more accessible. Streamlining the production process also aligns with environmental sustainability by reducing waste and energy consumption. This study quantifies the economic and environmental impacts of incorporating recent advances into end-to-end continuous bioprocessing of mAbs. The results demonstrate that, compared with an optimized best-in-class fed-batch process (with 15 g/l titer and multicolumn chromatography), continuous manufacturing can reduce the total annual production costs, facility footprint, plastic waste, and CO emissions by up to 23%, 51%, 57%, and 54%, respectively, in a multiproduct facility producing clinical and commercial lots. Additionally, uncertainty analysis indicates that these gains are even more substantial under demand fluctuations.
Rapid point-of-care pathogen sensing in the post-pandemic era
In the post-pandemic era, interest in on-site technologies capable of rapidly and accurately diagnosing viral or bacterial pathogens has significantly increased. Advances in functional nanomaterials and bioengineering have propelled the progress of point-of-care (POC) sensors, enhancing their speed, specificity, sensitivity, affordability, ease of use, and accuracy. Notably, biosensors that utilize surface-enhanced Raman scattering (SERS) technology have revolutionized the rapid and sensitive diagnosis of biomarkers in pathogenic infections. This review of current POC diagnostics highlights the growing emphasis on immunoassays for swift pathogen analysis, augmented by the integration of deep learning for swift interpretation of complex signals through tailored algorithms.
Designing synthetic microbial communities with the capacity to upcycle fermentation byproducts to increase production yields
Microbial cell factories, which convert feedstocks into a product of value, have the potential to help transition toward a bio-based economy with more sustainable ways to produce food, fuels, chemicals, and materials. One common challenge found in most bioconversions is the co-production, together with the product of interest, of undesirable byproducts or overflow metabolites. Here, we designed a strategy based on synthetic microbial communities to address this issue and increase overall production yields. To achieve our goal, we created a Yarrowia lipolytica co-culture comprising a wild-type (WT) strain that consumes glucose to make biomass and citric acid (CA), and an 'upcycler' strain, which consumes the CA produced by the WT strain. The co-culture produced up to two times more β-carotene compared with the WT monoculture using either minimal medium or hydrolysate. The proposed strategy has the potential to be applied to other bioprocesses and organisms.
A high-performance protein preparation approach in a single column-free step
Protein purification remains a formidable and costly technical obstacle in biotechnology. Here, we present a new column-free method, utilizing the cleavable self-aggregating tag 2.0 (cSAT2.0) scheme, to streamline protein production in Escherichia coli, yielding high quantities with exceptional purity. In shake-flask experiments using lysogeny broth (LB) medium, the cSAT2.0 scheme successfully produced one peptide and five proteins, with yields ranging from 24 mg/l to 89 mg/l, and purity levels exceeding 98%. The cSAT2.0 scheme also enabled high-throughput protein preparation on microplates. Furthermore, we scaled up the fermentation process for caplacizumab, achieving 1.4 g/l of highly purified protein in a 5-l fermenter. Our results demonstrate that the cSAT2.0 scheme can serve as an economical and robust platform for protein production from microplate to fermenter scales.
Revolutionizing IBD research with on-chip models of disease modeling and drug screening
Inflammatory bowel disease (IBD) comprises chronic inflammatory conditions with complex mechanisms and diverse manifestations, posing significant clinical challenges. Traditional animal models and ethical concerns in human studies necessitate innovative approaches. This review provides an overview of human intestinal architecture in health and inflammation, emphasizing the role of microfluidics and on-chip technologies in IBD research. These technologies allow precise manipulation of cellular and microbial interactions in a physiologically relevant context, simulating the intestinal ecosystem microscopically. By integrating cellular components and replicating 3D tissue architecture, they offer promising models for studying host-microbe interactions, wound healing, and therapeutic approaches. Continuous refinement of these technologies promises to advance IBD understanding and therapy development, inspiring further innovation and cross-disciplinary collaboration.
Intracellularly synthesized ssDNA for continuous genome engineering
Despite the prevalence of genome editing tools, there are still some limitations in dynamic and continuous genome editing. In vivo single-stranded DNA (ssDNA)-mediated genome mutation has emerged as a valuable and promising approach for continuous genome editing. In this review, we summarize the various types of intracellular ssDNA production systems and notable achievements in genome engineering in both prokaryotic and eukaryotic cells. We also review progress in the development of applications based on retron-based systems, which have demonstrated significant potential in molecular recording, multiplex genome editing, high-throughput functional variant screening, and gene-specific continuous in vivo evolution. Furthermore, we discuss the major challenges of ssDNA-mediated continuous genome editing and its prospects for future applications.
Cas9-PE: a robust multiplex gene editing tool for simultaneous precise editing and site-specific random mutation in rice
In molecular design breeding, the simultaneous introduction of desired functional genes through specific nucleotide modifications and the elimination of genes regulating undesired phenotypic traits or agronomic components require advanced gene editing tools. Due to limited editing efficiency, even with the use of highly precise editing tools, such as prime editing (PE), simultaneous editing of multiple mutation types poses a challenge. Here, we replaced Cas9 nickase (nCas9) with Cas9 to construct a Cas9-mediated PE (Cas9-PE) system in rice. This system not only enables precise editing, but also allows for site-specific random mutation. Moreover, leveraging the precision of Cas9-PE, we established a transgene-free multiplex gene editing system using a co-editing strategy. This strategy involved the Agrobacterium-mediated transient expression of the precise editing rice endogenous acetolactate synthase gene ALS to confer herbicide bispyribac-sodium (BS) resistance as a selection marker. This study provides a versatile and efficient multiplex gene editing tool for molecular design breeding.
Producing multiple chemicals through biological upcycling of waste poly(ethylene terephthalate)
Poly(ethylene terephthalate) (PET) waste is of low degradability in nature, and its mismanagement threatens numerous ecosystems. To combat the accumulation of waste PET in the biosphere, PET bio-upcycling, which integrates chemical pretreatment to produce PET-derived monomers with their microbial conversion into value-added products, has shown promise. The recently discovered Rhodococcus jostii RPET strain can metabolically degrade terephthalic acid (TPA) and ethylene glycol (EG) as sole carbon sources, and it has been developed into a microbial chassis for PET upcycling. However, the scarcity of synthetic biology tools, specifically designed for this non-model microbe, limits the development of a microbial cell factory for expanding the repertoire of bioproducts from postconsumer PET. Herein, we describe the development of potent genetic tools for RPET, including two inducible and titratable expression systems for tunable gene expression, along with serine integrase-based recombinational tools (SIRT) for genome editing. Using these tools, we systematically engineered the RPET strain to ultimately establish microbial supply chains for producing multiple chemicals, including lycopene, lipids, and succinate, from postconsumer PET waste bottles, achieving the highest titer of lycopene ever reported thus far in RPET [i.e., 22.6 mg/l of lycopene, ~10 000-fold higher than that of the wild-type (WT) strain]. This work highlights the great potential of plastic upcycling as a generalizable means of sustainable production of diverse chemicals.
A snapshot of biomanufacturing and the need for enabling research infrastructure
Biomanufacturing is crucial for the bioeconomy, with growing investment and attention from industries and governments. Over recent decades numerous biotech companies have been founded, and policies have increasingly prioritised sustainable production methods. However, translation of biotechnological innovations into industrial applications remains challenging, requiring interdisciplinary research infrastructures (RIs) to address gaps in bioprocess development, scalability, and competitiveness. This opinion examines the current landscape of biomanufacturing and highlights the pivotal role of RIs in supporting these transitions. It also proposes enhanced research interoperability, standardisation, and democratisation through meta-workflows that streamline operations within and between RIs. By improving data sharing, process harmonisation, and scalability, these ecosystems can help to overcome technical and economic barriers in a concerted effort towards sustainable, bio-based global manufacturing.
Plant-inspired building blocks for future plastics
The transition from a linear fossil-based economy to a renewable circular economy requires a new approach to produce building blocks for plastics. This provides opportunities to reshape the plastic landscape and will positively impact the wide range of applications that make use of plastics. We propose that plant enzymes, which underlie the large biochemical diversity present in plant specialized metabolism, will facilitate the production of novel building blocks for new polymers via biotechnological processes. Thereby, plant-inspired plastic building blocks may enable the development of new plastics for targeted applications that can contribute to a future with renewable plastics.
Chairside live biotherapeutic hydrogel for comprehensive periodontitis therapy
Periodontitis, characterized by microbial dysbiosis and immune dysregulation, destroys tooth-supporting tissues and negatively affects overall health. Current strategies face significant challenges in restoring damaged tissues while halting periodontitis progression. In this study, we introduce a live biotherapeutic product (LBP) in an engineered living hydrogel for comprehensive periodontitis therapy. A dental blue light-responsive hydrogel (LRG) was fabricated to deliver and confine live Lactobacillus rhamnosus GG (LGG) in periodontal pockets, endowing the LRG with sustained antibacterial and immunomodulatory effects. The LRG was engineered through peptide modification to also promote tissue regeneration. Both in vitro and in vivo evaluations confirmed the effectiveness of this integrated therapeutic strategy, which combines antibacterial, anti-inflammatory, and regenerative properties with an underlying immunomodulatory mechanism that involves suppressor of cytokine signaling (SOCS)3 upregulation and the Janus kinase/signal transducer and activator of transcription (JAK-STAT) pathway suppression in macrophages. Demonstrating a new paradigm, this proof of concept highlights the synergistic integration of live organisms and synthetic material engineering in a chairside treatment to address the multifaceted etiology of periodontitis.
Small cells with big photosynthetic productivities: biotechnological potential of the Picochlorum genus
The Picochlorum genus is a distinctive eukaryotic green-algal clade that is the focus of several current biotechnological studies. It is capable of extremely rapid growth rates and has exceptional tolerances to high salinity, intense light, and elevated temperatures. Importantly, it has robust stability and high-biomass productivities in outdoor field trials in seawater. These features have propelled Picochlorum into the spotlight as a promising model for both fundamental and biotechnological research. Recently, several genetic tools, including genome editing, were developed for these algae, enabling insights into Picochlorum photophysiology and algal transformations for expanded capabilities. Here, we survey the Picochlorum genus, its genetic toolbox, recently characterized transformants, and discuss the commercial potential of Picochlorum as a salt-water photoautotrophic biocatalyst.
Engineered, environmentally friendly leather-like bio-based materials
Leather is important to the global manufacturing industry, contributing to both the economy and society. However, because of ecological and ethical considerations, alternative bio-based materials to natural leather are now being investigated. Advancements in biotechnology and bio-based materials, combined with flourishing biomanufacturing, have driven product development. In recent years, animal-free, biotechnology-based leather-like material has seen significant growth. Recent progress in leather-like bio-based materials development has been achieved using proteins, mycelium, cellulose, and other sustainable natural materials. This review provides a comprehensive overview of these bio-based materials, addressing their challenges, practical implications, and potential to play a growing role in the emerging field of animal-free alternative. The development of 'future leather' has significant economic and environmental potential.
A cell-free bacteriophage synthesis system for directed evolution
Efficient phage production has always been an urgent need in fields such as drug discovery, disease treatment, and gene evolution. To meet this demand, we constructed a robust cell-free synthesis system for generating M13 phage by simplifying its genome, enabling a three-times faster efficiency compared with the traditional method in vivo. We further developed a cell-free directed evolution system in droplets, comprising a modified helper plasmid (ΔPS-ΔgIII-ΔgVI) and the simplified M13 genome-carrying gene mutation library. This system was greatly improved when coupled with fluorescence-activated droplet sorting (FADS). We successfully evolved the T7 RNA polymerase (RNAP), achieving a twofold higher activity to read through the T7 terminator. Moreover, we evolved the tryptophan tRNA into a suppressor tRNA with an eightfold increase in activity to read through the stop codon UAG.
Peptides from non-immune proteins target infections through antimicrobial and immunomodulatory properties
Encrypted peptides (EPs) have been recently described as a new class of antimicrobial molecules. They have been found in numerous organisms and have been proposed to have a role in host immunity and as alternatives to conventional antibiotics. Intriguingly, many of these EPs are found embedded in proteins unrelated to the immune system, suggesting that immunological responses extend beyond traditional host immunity proteins. To test this idea, we synthesized and analyzed representative peptides derived from non-immune human proteins for their ability to exert antimicrobial and immunomodulatory properties. Most of the tested peptides from non-immune proteins, derived from structural proteins as well as proteins from the nervous and visual systems, displayed potent in vitro antimicrobial activity. These molecules killed bacterial pathogens by targeting their membrane, and those originating from the same region of the body exhibited synergistic effects when combined. Beyond their antimicrobial properties, nearly 90% of the peptides tested exhibited immunomodulatory effects, modulating inflammatory mediators, such as interleukin (IL)-6, tumor necrosis factor (TNF)-α, and monocyte chemoattractant protein-1 (MCP-1). Moreover, eight of the peptides identified, collagenin-3 and 4, zipperin-1 and 2, and immunosin-2, 3, 12, and 13, displayed anti-infective efficacy in two different preclinical mouse models, reducing bacterial infections by up to four orders of magnitude. Altogether, our results support the hypothesis that peptides from non-immune proteins may have a role in host immunity. These results potentially expand our notion of the immune system to include previously unrecognized proteins and peptides that may be activated upon infection to confer protection to the host.
Leveraging the versatile properties of bacterial spores in materials
Inspired by biological functions of living systems, researchers have engineered cells as independent functional materials or integrated them within a natural or synthetic matrix to create engineered living materials (ELMs). However, the 'livingness' of cells in such materials poses serious drawbacks, such as a short lifespan and the need for cold-chain logistics. Bacterial spores have emerged as a game changer to bypass these shortcomings as a result of their intrinsic dormancy and resistance against harsh conditions. Emerging synthetic biology tools tailored for engineering spores and better understanding of their physical properties have led to novel applications of spore-based materials. Here, we review recent advances in such materials and discuss future challenges for the development of time- and cost-efficient spore-based materials with high performance.
Cellobiose lipids: applications, production, and downstream processing
Cellobiose lipids (CBLs) are glycolipid biosurfactants that have garnered attention due to their potential applications in diverse industries. Here, we review the current state of CBL research, from production and purification, to the potential applications of CBLs. We elucidate CBL functionality and consider some commercial applications, as well as how operating conditions (e.g., media and organism, or production approaches) impact productivity. Methodologies based on enzymatic synthesis or postproduction chemical modification of CBL variants are also presented. Given the importance of purity in current CBL applications, we discuss CBL separation and purification techniques. Finally, we highlight the importance of techno-economic and life-cycle assessments for the industrialisation of CBLs, while suggesting potential future routes for investigation.
In vitro human cell-based models: What can they do and what are their limitations?
Fabricating an advanced electrogenic chassis by activating microbial metabolism and fine-tuning extracellular electron transfer
Exploiting electrogenic microorganisms as unconventional chassis hosts offers potential solutions to global energy and environmental challenges. However, their limited electrogenic efficiency and metabolic versatility, due to genetic and metabolic constraints, hinder broader applications. Herein, we developed a multifaceted approach to fabricate an enhanced electrogenic chassis, starting with streamlining the genome by removing extrachromosomal genetic material. This reduction led to faster lactate consumption, higher intracellular NADH/NAD and ATP/ADP levels, and increased growth and biomass accumulation, as well as promoted electrogenic activity. Transcriptome profiling showed an overall activation of cellular metabolism. We further established a molecular toolkit with a vector vehicle incorporating native replication block and refined promoter components for precise gene expression control. This enabled engineered primary metabolism for greater environmental robustness and fine-tuned extracellular electron transfer (EET) for improved efficiency. The enhanced chassis demonstrated substantially improved pollutant biodegradation and radionuclide removal, establishing a new paradigm for utilizing electrogenic organisms as novel biotechnology chassis.