Biotechnology Advances

Oral thin films (OTFs) are innovative dosage forms that have gained tremendous attention for the delivery of nutraceuticals. They are ultra-thin, flexible sheets that can be easily placed on the tongue, sublingual or buccal mucosa (inner lining of the cheek). These thin films possess several advantages for nutraceutical delivery including ease of administration, rapid disintegration, fast absorption, rapid onset of action, bypass first-pass hepatic metabolism, accurate dosing, enhanced stability, portability, discreetness, dose flexibility and most importantly consumer acceptance...

L-Cysteine and L-methionine, as the only two sulfur-containing amino acids among the canonical 20 amino acids, possess distinct characteristics and find wide-ranging industrial applications. The use of different organisms for fermentative production of L-cysteine and L-methionine is gaining increasing attention, with Escherichia coli being extensively studied as the preferred strain. This preference is due to its ability to grow rapidly in cost-effective media, its robustness for industrial processes, the well-characterized metabolism, and the availability of molecular tools for genetic engineering...

Membraneless organelles (MLOs) formed by liquid-liquid phase separation (LLPS) have been extensively studied due to their spatiotemporal control of biochemical and cellular processes in living cells. These findings have provided valuable insights into the physicochemical principles underlying the formation and functionalization of biomolecular condensates, which paves the way for the development of versatile phase-separating systems capable of addressing a variety of application scenarios. Here, we highlight the potential of constructing synthetic MLOs with programmable and functional properties...

Thraustochytrids are marine microorganisms known for their fast growth and ability to store lipids, making them useful for producing polyunsaturated fatty acids (PUFAs), biodiesel, squalene, and carotenoids. However, the high cost of production, mainly due to expensive fermentation components, limits their wider use. A significant challenge in this context is the need to balance production costs with the value of the end products. This review focuses on integrating the efficient utilization of waste with Thraustochytrids fermentation, including the economic substitution of carbon sources, nitrogen sources, and fermentation water...

Carrageenan, the major carbohydrate component of some red algae, is an important renewable bioresource with very large annual outputs. Different types of carrageenolytic enzymes in the carrageenan metabolic pathway are potentially valuable for the production of carrageenan oligosaccharides, biofuel, and other chemicals obtained from carrageenan. However, these enzymes are not well-developed for oligosaccharide or biofuel production. For further application, comprehensive knowledge of carrageenolytic enzymes is essential...

Nitrilases represent a distinct class of enzymes that play a pivotal role in catalyzing the hydrolysis of nitrile compounds, leading to the formation of corresponding carboxylic acids. These enzymatic entities have garnered significant attention across a spectrum of industries, encompassing pharmaceuticals, agrochemicals, and fine chemicals. Moreover, their significance has been accentuated by mounting environmental pressures, propelling them into the forefront of biodegradation and bioremediation endeavors...

Inteins are proteins found in nature that execute protein splicing. Among them, split inteins stand out for their versatility and adaptability, presenting creative solutions for addressing intricate challenges in various biological applications. Their exquisite attributes, including compactness, reliability, orthogonality, low toxicity, and irreversibility, make them of interest to various fields including synthetic biology, biotechnology and biomedicine. In this review, we delve into the inherent challenges of using inteins, present approaches for overcoming these challenges, and detail their reliable use for specific cellular tasks...

As an energy-storage substance of microorganisms, polyhydroxybutyrate (PHB) is a promising alternative to petrochemical polymers. Under appropriate fermentation conditions, PHB-producing strains with metabolic diversity can efficiently synthesize PHB using various carbon sources. Carbon-rich wastes may serve as alternatives to pure sugar substrates to reduce the cost of PHB production. Genetic engineering strategies can further improve the efficiency of substrate assimilation and PHB synthesis. In the downstream link, PHB recycling strategies based on green chemistry concepts can replace PHB extraction using chlorinated solvents to enhance the economics of PHB production and reduce the potential risks of environmental pollution and health damage...

The extraordinary success that chimeric antigen receptor (CAR) T cell therapies have shown over the years on fighting hematological malignancies is evidenced by the six FDA-approved products present on the market. CAR T treatments have forever changed the way we understand cellular immunotherapies, as current research in the topic is expanding even outside the field of cancer with very promising results. Until now, virus-based strategies have been used for CAR T cell manufacturing. However, this methodology presents relevant limitations that need to be addressed prior to wide spreading this technology to other pathologies and in order to optimize current cancer treatments...

The rapid development of synthetic biology has significantly improved the capabilities of mono-culture systems in converting different substrates into various value-added bio-chemicals through metabolic engineering. However, overexpression of biosynthetic pathways in recombinant strains can impose a heavy metabolic burden on the host, resulting in imbalanced energy distribution and negatively affecting both cell growth and biosynthesis capacity. Synthetic consortia, consisting of two or more microbial species or strains with complementary functions, have emerged as a promising and efficient platform to alleviate the metabolic burden and increase product yield...

Mitigating greenhouse gas emissions is a critical challenge for promoting global sustainability. The utilization of CO2 and CH4 as substrates for the production of valuable products offers a promising avenue for establishing an eco-friendly economy. Biocatalysis, a sustainable process utilizing enzymes to facilitate biochemical reactions, plays a significant role in upcycling greenhouse gases. This review provides a comprehensive overview of the enzymes and associated reactions involved in the biocatalytic conversion of CO2 and CH4 ...

Genome engineering has revolutionized several scientific fields, ranging from biochemistry and fundamental research to therapeutic uses and crop development. Diverse engineering toolkits have been developed and used to effectively modify the genome sequences of organisms. However, there is a lack of extensive reviews on genome engineering technologies based on mobile genetic elements (MGEs), which induce genetic diversity within host cells by changing their locations in the genome. This review provides a comprehensive update on the versatility of MGEs as powerful genome engineering tools that offers efficient solutions to challenges associated with genome engineering...

Biohydrogen (BioH2 ) is widely recognized as a sustainable and environmentally friendly energy source, devoid of any detrimental impact on the environment. Lignocellulosic biomass (LB) is a readily accessible and plentiful source material that can be effectively employed as a cost-effective and sustainable substrate for Bio-H2 production. Despite the numerous challenges, the ongoing progress in LB pretreatment technology, microbial fermentation, and the integration of molecular biology techniques have the potential to enhance Bio-H2 productivity and yield...

Defects in the genome cause genetic diseases and can be treated with gene therapy. Due to the limitations encountered in gene delivery, lipid-based supramolecular colloidal materials have emerged as promising gene carrier systems. In their non-functionalized form, lipid nanoparticles often demonstrate lower transgene expression efficiency, leading to suboptimal therapeutic outcomes, specifically through reduced percentages of cells expressing the transgene. Due to chemically active substituents, the engineering of delivery systems for genetic drugs with specific chemical ligands steps forward as an innovative strategy to tackle the drawbacks and enhance their therapeutic efficacy...

Most enzyme modification strategies focus on designing the active sites or their surrounding structures. Interestingly, a large portion of the enzymes (60%) feature active sites located within spacious cavities. Despite recent discoveries, cavity-mediated enzyme engineering remains crucial for enhancing enzyme properties and unraveling folding-unfolding mechanisms. Cavity engineering influences enzyme stability, catalytic activity, specificity, substrate recognition, and docking. This article provides a comprehensive review of various cavity engineering models for enzyme modification, including cavity creation, filling, and reshaping...

Transcriptional regulators generate connections between biological signals and genetic outputs. They are used robustly for sensing input signals in building genetic circuits. However, each regulator can only generate a fixed connection, which generates constraints in linking multiple signals for more complex processes. Recent studies discovered that a domain swapping strategy can be applied to various regulator families to create modular regulators for new signal-output connections, significantly broadening possibilities in circuit design...

As a sustainable and renewable alternative to petroleum fuels, advanced biofuels shoulder the responsibility of energy saving, emission reduction and environmental protection. Traditional engineering of cell factories for production of advanced biofuels lacks efficient high-throughput screening tools and regulating systems, impeding the improvement of cellular productivity and yield. Transcription factor-based biosensors have been widely applied to monitor and regulate microbial cell factory products due to the advantages of fast detection and in-situ screening...

Lignocellulosic biomass holds a crucial position in the prospective bio-based economy, serving as a sustainable and renewable source for a variety of biobased products. These products play a vital role in displacing fossil fuels and contributing to environmental well-being. However, the inherent recalcitrance of biomass poses a significant obstacle to the efficient access of sugar polymers. Consequently, the bioconversion of lignocellulosic biomass into fermentable sugars remains a prominent challenge in biorefinery processes to produce biofuels and biochemicals...

Non-homogeneous enzyme-catalyzed systems are more widely used than homogeneous systems. Distinguished from the conventional biphasic approach, Pickering emulsion stabilized by ultrafine solid particles opens up an innovative platform for biocatalysis. Their vast specific surface area significantly enhances enzyme-substrate interactions, dramatically increasing catalytic efficiency. This review comprehensively explores various aspects of Pickering emulsion biocatalysis, provides insights into the multiple types and mechanisms of its catalysis, and offers strategies for material design, enzyme immobilization, emulsion formation control, and reactor design...

The global industrialization and modernization have witnessed a rapid progress made in agricultural production, along with the issue of soil heavy metal (HM) pollution, which has posed severe threats to soil quality, crop yield, and human health. Phytoremediation, as an alternative to physical and chemical methods, offers a more cost-effective, eco-friendly, and aesthetically appealing means for in-situ remediation. Despite its advantages, traditional phytoremediation faces challenges, including variable soil physicochemical properties, the bioavailability of HMs, and the slow growth and limited biomass of plants used for remediation...