Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P(3HB-co-4HB) or P34HB) is a promising biopolyester for applications in food packaging, medical sutures, drug delivery, and tissue engineering due to its tunable thermomechanical properties. However, industrial-scale production of P34HB from glucose remains challenging. In this study, scalable P34HB production by engineered Halomonas bluephagenesis was

Alkanes are considered among the most promising candidates for next-generation biofuels. Amongst various pathways discovered for alkane production, the cyanobacterial AAR (acyl ACP reductase) - ADO (aldehyde deformylating oxygenase) pathway has been the most studied pathway. Considering that cyanobacteria have the innate ability to produce alkanes, they can serve as an excellent chassis for sustainable

2-Phenylethanol (2-PE) is a natural aromatic compound with properties that make it a potential biological oxygenate for petroleum-derived gasoline. In plants, 2-PE biosynthesis competes with the phenylpropanoid pathway for the common precursor, phenylalanine (Phe). The phenylpropanoid pathway directs significant carbon flux towards the formation of lignin, a major biopolymer in plant cell walls that

Itaconic acid, a versatile platform chemical, has garnered significant attention due to its broad use in polymers, resins, and bio-based materials. Although fungi, especially Aspergillus and Ustilago, are the main producers of itaconic acid, reprogramming yeast species like Saccharomyces cerevisiae and Yarrowia lipolytica as alternative production platforms offers advantages for industrial bioproduction

Malonyl-CoA is a central precursor involved in the synthesis of various bio-based chemicals, including polyketides, fatty acids, and flavonoids. However, the production of these chemicals is often limited by the availability of malonyl-CoA. Based on retrosynthesis principles, we designed two thermodynamically favorable malonyl-CoA pathways using L-glutamate and L-aspartate as substrates. The novel

Pseudomonas putida strains are prized biocatalysts, renowned for their versatility in degrading diverse chemicals, tolerating organic solvents, and withstanding environmental stressors. Central to their adaptive success is the precise regulation of primary carbon metabolism, with HexR emerging as a key regulator. While previous research has explored HexR binding through in vitro assays and comparative

3-Hydroxypropionic acid (3-HP) is a versatile platform chemical with broad applications, serving as a precursor for the synthesis of value-added chemicals as well as the biodegradable polymers. However, current industrial production of 3-HP relies on chemical synthesis, which requires harmful raw materials and harsh reaction conditions. As a sustainable alternative, microbial biosynthesis of 3-HP has

The shikimate pathway is a crucial metabolic route for the biosynthesis of numerous valuable chemicals. In this study, we engineered the shikimate pathway in plants via expression of microbial enzymes to produce the two important antioxidants gallate and arbutin. The engineered pathways utilize the aromatics protocatechuate and 4-hydroxybenzoate as metabolic intermediates. Through transient expression

The oleaginous yeast, Yarrowia lipolytica has gained interest as a biotechnological chassis to produce foods, chemicals, pharmaceuticals, and biofuels. To reduce production costs and sustainability, inexpensive and abundant feedstocks such as lignocellulose must be used for bioproduction. Since lignocellulosic biomass contains components that cannot be utilised by Y. lipolytica, it is important to

The reductive tricarboxylic acid (rTCA) cycle is a crucial metabolic pathway employed in the microbial production of C4-dicarboxylic acids, especially succinic acid (SA). However, the inherent redox constraints associated with this cycle pose significant limitations on the yields of SA. Here, we address this critical bottleneck by engineering a non-canonical reductive TCA (Nc-rTCA) pathway in oleaginous

CRISPR interference (CRISPRi) has emerged as a valuable tool for redirecting metabolic flux to enhance bioproduction. However, its application is often constrained by two challenges: (i) rationally identifying effective gene targets for downregulation and (ii) efficiently constructing multiplexed CRISPRi systems. In this study, we address both challenges by integrating a computational prioritization

Psychedelic-assisted therapy is emerging as a highly promising approach for treating depression, with psilocybin, a psychoactive compound in magic mushrooms, gaining the most recognition for its efficacy in treating post-traumatic stress disorder and treatment-resistant depression. However, its low natural abundance makes extraction costly, necessitating alternative production methods. While engineered

Human milk fats (HMFs) could facilitate nutrient absorption in the infant gut, with 1,3-olein-2-palmitin (OPO) and 1-olein-2-palmitin-3-linolein (OPL) being the most abundant components. The construction of microbial cell factories has garnered significant interest due to their potential to synthesize HMFs from cheap raw materials. However, the substrate preference of endogenous triglyceride (TAG)

Free fatty acid (FFA) production in bacteria is a key target for metabolic engineering. The knockout of the acyl-ACP synthetase (AAS) prevents reincorporation of FFA into the fatty acid biosynthetic cycle and is widely used to enhance their secretion. However, the role of AAS in membrane lipid remodeling under environmental stress, such as altered temperature, remains poorly understood. In cyanobacteria

Microbial natural products (NPs) are a pivotal reservoir for drugs used in human health and agriculture. Andrimid, a polyketide-non-ribosomal peptide hybrid antibiotic inhibiting bacterial acetyl-CoA carboxylase, shows enormous potential in antibiotic drug development to mitigate antimicrobial resistance. However, industrial-scale manufacturing and downstream development of andrimid are largely prohibited

Propionic acid is a key three carbon platform chemical with broad applications in food preservation, pharmaceuticals, and polymer production. Traditional microbial production of propionic acid employing Propionibacterium species is constrained by slow growth, and limited genetic engineering tools, thereby restricting its industrial use. Here, we report the development of a novel biosynthetic pathway

Lacto-N-tetraose (LNT), an important human milk oligosaccharide with prebiotic benefits, was successfully produced de novo in Bacillus subtilis, establishing this Generally Recognized as Safe (GRAS) organism as a suitable platform for infant nutritional ingredients. A detailed enzyme screening identified three key enzymes: β-1,3-galactosyltransferase from Pseudogulbenkiania ferrooxidans, β-1,3-N-a

The high-value sesquiterpenoid (+)-nootkatone has important applications in food, agriculture, and pharmaceutical industries. Extraction from plant material, however, is technically challenging and inefficient due to inherent low concentrations in native sources. Over the last decade, the eukaryotic green microalga Chlamydomonas reinhardtii has emerged as a powerful alternative for heterologous terpenoid

In a circular bioeconomy, the one-way conversion of petroleum to chemicals and CO2 is replaced with processes that reduce CO2 to energy carriers and useful materials that are returned to CO2 upon combustion. A circular bioeconomy that relies on photosynthesis to generate sugars as the chief energy carrier and precursor to chemical building blocks has yet to overcome many recalcitrant aspects of plant-based

Ammonia, a byproduct of glutamine metabolism, inhibits cell growth and reduces product yield and quality in mammalian cell culture. To identify novel genes associated with ammonia resistance, a genome-wide CRISPR knockout screening was conducted in monoclonal antibody (mAb)-producing human embryonic kidney 293 (HEK-mAb) cells using a virus-free, recombinase-mediated cassette exchange-based gRNA interrogation

Bacillus subtilis is a GRAS-certified chassis, yet scalable and precise multi-gene regulation remains a bottleneck in its metabolic engineering. Here, we present bsBETTER, a base editor-guided, template-free system enabling high-diversity expression tuning across multiple genomic loci. By editing ribosome binding sites (RBSs) of 12 lycopene biosynthetic genes, we generated thousands of combinatorial

Product yields for biomanufacturing processes are often constrained by the tight coupling of cellular energy generation and carbon metabolism in sugar-based fermentation systems. To overcome this limitation, we engineered Escherichia coli to utilize hydrogen gas (H2) and formate (HCOO−) as alternative sources of energy and reducing equivalents, thereby decoupling energy generation from carbon metabolism

The transition towards a sustainable bioeconomy requires the use of alternative feedstocks, with CO2-derived formate emerging as a promising candidate for industrial biotechnology. Despite its beneficial characteristics as a feedstock, microbial assimilation of formate is limited by the inefficiency of naturally evolved formate-fixing pathways. To overcome this limitation, synthetic formate reduction

Efficient utilization of pentose sugars is critical for advancing sustainable biomanufacturing using lignocellulose. However, many host strains capable of consuming glucose and lignin-derived monomers are unable to utilize pentose sugars. Here, we engineered Acinetobacter baylyi ADP1 for the utilization of D-xylose and L-arabinose. We first modelled different pentose utilization pathways using flux

Vanillin is an aromatic flavor compound widely used in the food, pharmaceutical, and cosmetic industries. Microbial biosynthesis offers a sustainable alternative to traditional plant extraction and chemical synthesis; however, the susceptibility of vanillin to redox reactions and the weak enzyme activity in cells severely limit the vanillin production capacity by microbial biosynthesis. This study

Caffeic acid (CA) is a valuable phenolic compound with wide applications in pharmaceuticals, food additives, and materials. However, its microbial production faces several challenges, including low heterologous enzyme activity and product toxicity. Here, we report the development of an integrated biosensor-driven high-throughput screening (HTS) platform for the efficient production of CA in Escherichia

By 2050, the global population is projected to reach 9.7 billion, necessitating a 70 % increase in traditional agricultural output to meet growing demands. However, critical constraints are emerging as arable land and water resources approach their sustainable utilization thresholds. In this context, ensuring safe, efficient, and sustainable food production has become a pivotal issue intertwined with

Efficiently rewiring microbial metabolism for molecule production lies at the core of industrial metabolic engineering. Combinatorial libraries are useful for directing metabolism towards molecule production; however, their construction is labor-intensive, and their use in iterative strain engineering campaigns is often restricted by site-specific genomic integration. Here we present an automation-friendly

D-Pantothenate (D-PA), a crucial precursor for coenzyme A, is widely used in various industries. Traditional chemical synthesis of D-PA involves toxic inputs, including cyanide, and generates environmental pollution. Total biosynthesis from glucose still has limitation in long reaction time and low titer. To provide a new approach to D-PA, we designed a hybrid pathway. First, we used green chemical

Inducible genetic operation systems constitute essential tools in microbial synthetic biology and metabolic engineering. However, inducible systems in non-model microbes, particularly thermophiles, are rarely reported. Acetivibrio thermocellus (previously termed Clostridium thermocellum), a representative strain of thermophilic non-model microbes, currently serves as a promising chassis organism in

L-valine is an essential amino acid widely used in the food, pharmaceutical, and animal feed industries. Currently, engineering microbial cell factories to produce L-valine from low-cost feedstocks has emerged as a leading strategy. However, there is still a lack of an L-valine-producing strain that simultaneously exhibits high titer, high yield, and high productivity. The metabolic engineering strategies

L-arginine is a high-value amino acid with widely utilized in the food, feed, and pharmaceutical industries. However, its large-scale biosynthesis remains limited by the low efficiency of current microbial strains. In this study, intracellular citrulline accumulation in Escherichia coli-Arg4 was enhanced by 2.45-, 1.90-, and 1.94-fold through supplementation with monosodium glutamate, monosodium aspartate

Cytidine-5′-diphosphocholine (CDP-choline) is a crucial neuroprotective agent. Current industrial production relies on chemical and enzymatic methods that face inherent sustainability challenges and share a dependence on the costly precursor, cytidine monophosphate (CMP). Here, we report the systems metabolic engineering of Bacillus subtilis for the efficient, de novo biosynthesis of CDP-choline from

The budding yeast Saccharomyces cerevisiae is one of the most widely used microbial cell factories for heterologous protein and enzyme production. However, improving production efficiency and tailoring enzyme properties remain a major challenge. Here we identified MDG1, a gene involved in the pheromone signaling pathway, as a previously unrecognized regulator that significantly enhances cellulase production

Clostridium thermocellum is an increasingly well-studied organism with considerable advantages for consolidated bioprocessing towards ethanol production. Here, a genome-scale resource balance analysis (RBA) model of C. thermocellum, ctRBA, is reconstructed based on a recently published stoichiometric model (iCTH669), global proteomics, and 13C MFA datasets to analyze proteome allocation and the burden

1,3-Propanediol (1,3-PDO) is an essential monomer used in the synthesis of polytrimethylene terephthalates. However, the microbial production of 1,3-PDO is limited by product tolerance and Vitamin B12 (VB12) supplementation. In this study, FMME-KP—a Klebsiella pneumoniae strain with a 1,3-PDO titer of 63.4 g/L. Through pathway reprogramming, the 1,3-PDO titer of strain FMME-14 was increased by 49.1 %

Pyridine carboxylic acids, because of their structural similarity to aromatic carboxylic acids, have garnered increasing attention as alternative compounds in chemical synthesis. However, their broader utilization has been limited by challenges in biosynthetic production. In this study, we developed a metabolic pathway for biosynthesizing 2,5-pyridinedicarboxylate (2,5-PDCA) from glucose from p-aminobenzoate

Oleaginous yeasts are used commercially to produce oleochemicals and hold potential also for biodiesel production. In response to nitrogen or phosphorous limitation, oleaginous yeasts accumulate lipids in the form of triacylglycerols. Previous work has investigated potential mechanisms by which nutrient limitation induces lipid biosynthesis without verifying whether lipid biosynthesis flux is actually

Poly(3-hydroxyoctanoate) (PHO) is a medium-chain-length PHA with low crystallinity and high elongation to break ratio, unlike the brittle short-chain-PHAs like PHB. These properties make PHO a promising candidate for industrial and biomedical applications. In this study, we demonstrated the production of PHO in Escherichia coli from a renewable and inexpensive glycerol feedstock by engineering fatty

Secondary metabolites have crucial medicinal and industrial applications, but their alignment with primary metabolism remains unclear. As secondary metabolism depends on primary metabolism for precursor supply, we present a pan-reactome analysis of 242 Streptomyces strains to investigate their association and disconnection. This analysis includes phylogenetic grouping of the strains using genome data

The exploitation of Vibrio natriegens as an unconventional host for biotechnology has progressed rapidly. This development is not only a result of the remarkable high growth rate of this marine bacterium on different substrates but is also possible due to good handling properties, a versatile metabolism and its inherent natural competence – features that have facilitated the development of a sophisticated

The biopharmaceutical sector relies on CHO cells to investigate biological processes and as the preferred host for production of biotherapeutics. Simultaneously, advancements in CHO cell genome assembly have provided insights for developing sophisticated genetic engineering strategies. While the majority of these efforts have focused on coding genes, with some interest in transcribed non-coding RNAs

Monosaccharides exist in either “D” or “L” conformations, with L-sugars being much less abundant in nature and therefore classified as “rare sugars.” Rare sugars hold significant potential due to their unique interactions with biological systems, offering health, food, and crop benefits. One such sugar, L-sorbose, serves as a critical precursor to Vitamin C and offers a low-calorie, moderately sweet

Lignocellulosic biomass represents a promising renewable feedstock for sustainable biochemical production, with p-hydroxycinnamates emerging as key aromatic building blocks derived from agricultural residues and grassy plants. C. glutamicum has recently been engineered to produce cis, cis-muconate (MA), a high-value platform chemical used in biobased plastics, resins, and specialty chemicals. However

D-pantothenic acid (D-PA) is a vital water-soluble vitamin with diverse industrial applications, driving the demand for efficient microbial production. Here, we rationally engineered an Escherichia coli strain to enhance D-PA production through metabolic engineering. First, to enhance carbon utilization efficiency, competing byproduct pathways were deleted and the pentose phosphate pathway was downregulated

Adenosine triphosphate (ATP) regeneration by substrate-level phosphorylation is a general feature of cancer metabolism, even under normoxic conditions (aerobic glycolysis). However, it is unclear why cancer cells prefer inefficient aerobic glycolysis over the highly efficient process of oxidative phosphorylation for ATP regeneration. To investigate the metabolic principles underlying aerobic glycolysis

Harnessing the potential of tailoring enzymes within fungal natural product (NP) biosynthetic gene clusters (BGCs) can significantly enhance NP diversity and production efficiency via artificially constructed microbial cell factories. To achieve this, an efficient genome mining method is crucial, especially since the functions of many putative enzymes in databases are unknown. As a test case, we aimed

Cyanobacteria are capable of fixing CO2 using sunlight as the sole energy source and are promising microbial platforms for sustainable bioproduction of fuels, commodity chemicals, food and pharmaceuticals. L-lysine is an essential amino acid to humans and animals and is a precursor to synthesis of building blocks for nylon and polyesters. Its industrial production is currently solely based upon fermenting

The design of synthetic driving forces for biosynthetic pathway is crucial for directing carbon flux toward the target product. Optimizing cellular redox status is one of the key strategies for constructing microbial cell factories. In this study, we attempt to create a novel redox imbalance force-driven (RIFD) strategy to direct carbon flow toward the target synthetic pathway. Initially, we increased

D-panthenol is a compound of significant importance in the pharmaceutical, cosmetic, and nutraceutical sectors, attributed to its remarkable moisturizing, anti-inflammatory, and tissue repair properties. Traditional chemical synthesis encounters several challenges, including the generation of toxic by-products, low enantiomeric excess, and expensive purification processes. To date, complete biosynthesis

Light-driven microbial communities consisting of phototrophs and heterotrophs represent an emerging frontier for biochemicals production from carbon dioxide (CO2). However, the construction of stable and robust light-driven artificial microbial communities remains challenging because the dominant strain wins the competition for nutrient and leads to the instability of subpopulations. Inspired by natural

Many herbivorous insects feed on unbalanced diets and rely on bacterial endosymbionts to meet all their nutritional needs. This is the case for the pea aphid (Acyrthosiphon pisum), a plant pest whose remarkable growth and reproductive capacities cannot be sustained by its sole nutritional resource, the plant phloem sap, and which relies on a symbiotic relationship maintained over millions of years

Activating silent biosynthetic gene clusters (BGCs) within various microorganisms is an important approach to uncover valuable natural products. In this study, we reported the capture and activation of two large silent BGCs from a marine-derived Streptomyces sp. SCSGAA 0027 in the heterologous host Streptomyces albus J1074 by inserting a widely used constitutive promoter kasOp∗ upstream of the core

ε-Poly-L-lysine (ε-PL) is a homopolymer of L-lysine residues produced by microorganisms, widely utilized in the food, pharmaceutical, and cosmetic industries. However, the development of efficient microbial cell factories (MCFs) for ε-PL production remains challenging. In this study, L-lysine importers were systematically screened, identified, and engineered to enhance ε-PL biosynthesis. First, an