Valuable Natural Products Research Articles

Overproduction of Cucurbitadienol through Modular Metabolic Engineering and Fermentation Optimization in Saccharomyces cerevisiae.

Cucurbitadienol is a key intermediate in the biosynthesis of cucurbitane-type compounds and serves as a precursor for mogrosides, cucurbitacins, and other valuable natural products of potential biological and food importance. However, microbial fermentation for cucurbitadienol production remains inefficient, limiting its potential for further industrial application. This study achieved the efficient synthesis of cucurbitadienol through a multimodular strategy. First, an N-degron tag was used to direct metabolic flux toward cucurbitadienol synthesis without compromising cell growth. Second, enzyme engineering strategies were employed to improve the utilization efficiency of intermediate metabolites. Finally, to increase precursor availability, the transcription factor UPC2-1 was introduced, which upregulated the expression of ERGs in the pre-squalene pathway. After eliminating nitrogen supplementation and optimizing fermentation conditions, cucurbitadienol accumulation in the 5 L bioreactor increased to 6.1 g/L, representing the highest titer reported to date. These findings provide a solid foundation for the industrial-scale production of cucurbitadienol and its derivatives.

Comparison of Different High-Speed Countercurrent Chromatography Injection Modes for the Separation of Glabridin.

In this work, different injection modes (single injection, overlapping injection, and continuous injection) of countercurrent chromatography were used and compared for the isolation and purification of glabridin from the crude extract of Glycyrrhiza glabra. The two-phase solvent system consisting of n-hexane/ethyl acetate/methanol/water (5:4:5:4, v/v) was employed as stationary and mobile phases. Compared with single injection mode, both overlapping injection and continuous injection modes exhibited higher separation efficiency at the same sample loading capacity. Their total separation times were 89.68% and 53.97% of the single injection mode, and the total solvent consumption were 74.88% and 52.49% of the single injection mode, respectively. Meanwhile, the yield, purity, and recovery rate of these three injection modes were almost the same. It was demonstrated that high-speed countercurrent chromatography in continuous injection mode was the most effective way for separating glabridin in three methods and it can be further applied to the separation of other economically valuable natural products.

Gene-Directed In Vitro Mining Uncovers the Insect-Repellent Constituent from Mugwort (Artemisia argyi).

Plants contain a vast array of natural products yet to be discovered, particularly those minor bioactive constituents. Identification of these constituents requires a significant amount of plant material, presenting considerable technical challenges. Mugwort (Artemisia argyi) is a widely recognized insect repellent herb, particularly renowned for its extensive usage during the Dragon Boat Festival in China, but the specific constituent responsible for its repellent activity remains unknown. Here, we employed a gene-directed in vitro mining approach to characterize mugwort terpene synthases (TPSs) systematically in a yeast expression system. Based on the establishment of "Terpene synthase-standard library", we have successfully identified 54 terpene products, including a novel compound designated as cyclosantalol. Through activity screening, we have identified that (+)-intermedeol, which presents in trace amount in plants, exhibits significant repellent activity against mosquitoes and ticks. After establishing its safety and efficacy, we then achieved its biosynthetic production in a yeast chassis, with an initial yield of 2.34 g/L. The methodology employed in this study not only identified a highly effective, safe, and commercially viable insect repellent derived from mugwort but also holds promise for uncovering and producing other valuable plant natural products in future research endeavors.

Structure of a Putative Terminal Amidation Domain in Natural Product Biosynthesis.

Bacteria are rich sources of pharmaceutically valuable natural products, many crafted by modular polyketide synthases (PKS) and non-ribosomal peptide synthetases (NRPS). PKS and NRPS systems typically contain a thioesterase (TE) to offload a linear or cyclized product from a carrier protein, but alternative chemistry is needed for products with a terminal amide. Several pathways with amidated products also possess an uncharacterized 400-amino acid terminal domain. We present the characterization and structure of this putative terminal amidation domain (TAD). TAD binds NAD with the nicotinamide near an invariant cysteine that is also accessible to an intermediate on a carrier protein, indicating a catalytic role. The TAD structure resembles cyanobacterial acyl-ACP reductase (AAR), which binds NADPH near an analogous catalytic cysteine. Bioinformatic analysis reveals that TADs are broadly distributed across bacterial phyla and often occur at the end of terminal NRPS modules, suggesting many amidated products may yet be discovered.

Discovery of a novel methionine biosynthetic route via O-phospho-l-homoserine.

Methionine (Met), a sulfur-containing amino acid, is essential for the underlying biological processes in living organisms. In addition to its importance as a starting building block for peptide chain elongation in protein biosynthesis, Met is a direct precursor of S-adenosyl-l-methionine, an indispensable methyl donor molecule in primary and secondary metabolism. Streptomyces bacteria are well known to produce diverse secondary metabolites, but many strains lack canonical Met pathway genes for l-homocysteine, a direct precursor of Met in bacteria, plants, and archaea. Here, we report the identification of a novel gene (metM) responsible for the Met biosynthesis in Streptomyces strains and demonstrate the catalytic function of the gene product, MetM. We further identified the metO gene, a downstream gene of metM, and showed that it encodes a sulfur-carrier protein (SCP). In in vitro analysis, MetO was found to play an important role in a sulfur donor by forming a thiocarboxylated SCP. Together with MetO (thiocarboxylate), MetM directly converted O-phospho-l-homoserine to l-homocysteine. O-Phospho-l-homoserine is also known as an intermediate for threonine biosynthesis in bacteria and plants, and MetM shares sequence homology with threonine synthase. Our findings thus revealed that MetM seizes O-phospho-l-homoserine from the threonine biosynthetic pathway and uses it as an intermediate of the Met biosynthesis to generate the sulfur-containing amino acid. Importantly, this MetM/MetO pathway is highly conserved in Streptomyces bacteria and distributed in other bacteria and archaea.IMPORTANCEMethionine (Met) is a sulfur-containing proteinogenic amino acid. Moreover, Met is a direct precursor of S-adenosyl-l-methionine, an indispensable molecule for expanding the structural diversity of natural products. Because Met and its derivatives benefit humans, the knowledge of Met biosynthesis is important as a basis for improving their fermentation. Streptomyces bacteria are well known to produce diverse and valuable natural products, but many strains lack canonical Met pathway genes. Here, we identified a novel l-homocysteine synthase (MetM) in Streptomyces and demonstrated that it converts O-phospho-L-homoserine to l-homocysteine using a thiocarboxylated sulfur-carrier protein as a sulfur donor. Since the metM is distributed in other bacteria and archaea, our pioneering study contributes to understanding Met biosynthesis in these organisms.

Natural synergy: Oleanolic acid-curcumin co-assembled nanoparticles combat osteoarthritis

Curcumin (Cur) is a natural polyphenol that is one of the most valuable natural products. However, its use as a functional food is limited by low water solubility, chemical instability and poor bioavailability. In this study, a supramolecular co-assembly strategy was used to construct an oleanolic acid-curcumin (OLA-Cur) co-assembly composite nano-slow-release treatment system. As a co-assembled compound, OLA is a widely present pentacyclic triterpenoid compound with multiple biological activities in the plant kingdom, which is expected to jointly alleviate the damaging effects of papain-induced mouse osteoarthritis model. The OLA-Cur NPs shows the solid core-shell structure, which can effectively improve the water solubility of Cur and OLA, and has good stability and sustained release characteristics. The analysis results show that the two compounds are mainly assembled through hydrogen bonding interactions, hydrophobic interactions, and π - π stacking interactions. The OLA-Cur NPs can inhibit the release of pro-inflammatory cytokines TNF-α, IL-6, and IL-1β induced by LPS in RAW264.7 mouse macrophages, promote the secretion of anti-inflammatory cytokine IL-10, and improve the oxidative stress index of hydrogen peroxide induced human rheumatoid arthritis synovial fibroblasts. In addition, it has a certain improvement effect on cartilage and subchondral bone damage in mouse osteoarthritis models. These findings suggest that constructing co-assembled composite nanoparticles based on pure natural compounds may break through the limitations of a variety of important nutritional ingredients in functional foods.

Evaluating the effectiveness of DNA-based methods to detect mislabelling and adulteration of chestnut honey

Honey is a naturally sweet and viscous substance that is consumed worldwide for its specific flavour, taste and biological properties, which in turn depend on the type of plant from which bees collect nectar. The quality of honey and, in particular, the information on the product label determines consumer choice and market price. In this study, the true botanical composition of chestnut monofloral honeys produced in Calabria (South Italy) was determined by DNA barcoding, PCR-RFLP and LAMP approaches, all based on the chloroplast trnL-trnF intergenic spacer region. All three DNA-based techniques were reliable in unambiguously identifying the floral composition of the different honeys tested at species level. However, the LAMP assay has the advantage over DNA barcoding and PCR-RFLP of greatly simplifying and speeding up the identification of Castanea sativa as the dominant plant species of the Calabrian chestnut honeys. This may encourage the adoption of this method by the food industry for the rapid detection of intentionally or accidentally mislabelled ingredients in this valuable natural product, thus ensuring consumer safety and protection.

Synthesis of N1-caffeoyl-N10-dihydrocaffeoylspermidine (Scotanamine D)

Abstract: N1-caffeoyl-N10-dihydrocaffeoylspermidine (Scotanamine D), a spermidine alkaloid isolated from various plants, is a medicinally valuable natural product. Recent studies have pointed out several health benefits of this compound. However, its synthetic procedures are still not described in the literature. We report the synthesis of this compound following two different schemes comprising multiple steps with excellent overall yields, which are 57% and 81%, respectively. These two synthetic schemes, which use commercially available and cheaper starting materials, can facilitate the large-scale manufacturing of Scotanamine D.

Elderberry Hydrolate: Exploring Chemical Profile, Antioxidant Potency and Antigenotoxicity for Cosmetic Applications

Elderberry (Sambucus nigra L.) hydrolate, derived from domestic steam distillation, holds promise as a multifunctional ingredient for skincare and cosmetic applications. This study investigates the chemical composition and biological activities of elderberry hydrolate obtained through steam distillation. Despite the growing interest in elderberry hydrolate, there has been a lack of comprehensive studies elucidating its chemical composition and potential bioactive constituents. To address this gap, we conducted a detailed analysis of elderberry hydrolate’s composition, antioxidant activity, and antigenotoxicity. Genotoxic evaluation and antioxidant assays (ABTS, DPPH) were conducted to assess its biological properties. We obtained elderberry hydrolate with a notable transfer of aromatic compounds through the steam distillation process, highlighting its efficacy and sustainability. The chemical characterization identified vital compounds, including phenylacetaldehyde, 2-acetyl-pyrrole, and an unidentified major component, collectively contributing to the hydrolate’s aromatic and biological properties. The genotoxic evaluation using the Comet assay demonstrated the hydrolate’s protective effects against DNA damage induced by hydrogen peroxide and streptonigrin. The optimal DNA protection was observed at 10% (w/v), attributed to the antioxidant activity of the identified compounds. The hydrolate exhibited significant antioxidant potential, demonstrating concentration-dependent responses and correlating higher concentrations with increased antioxidant activity. These findings underscore the multifaceted attributes of elderberry hydrolate, positioning it as a promising natural ingredient for skincare. This study supports elderberry hydrolate as a valuable natural and sustainable product development resource.

Volatiles emitted by Pseudomonas aurantiaca ST-TJ4 trigger systemic plant resistance to Verticillium dahliae

Verticillium dahliae is among the most devastating fungal pathogens, causing significant economic harm to agriculture and forestry. To address this problem, researchers have focused on eliciting systemic resistance in host plants through utilizing volatile organic compounds (VOCs) produced by biological control agents. Herein, we meticulously measured the quantity of V. dahliae pathogens in plants via RTqPCR, as well as the levels of defensive enzymes and pathogenesis-related (PR) proteins within plants. Finally, the efficacy of VOCs in controlling Verticillium wilt in cotton was evaluated. Following treatment with Pseudomonas aurantiaca ST-TJ4, the expression of specific VdEF1-α genes in cotton decreased significantly. The incidence and disease indices also decreased following VOC treatment. In cotton, the salicylic acid (SA) signal was strongly activated 24 h posttreatment; then, hydrogen peroxide (H2O2) levels increased at 48 h, and peroxidase (POD) and catalase (CAT) activities increased to varying degrees at different time points. The malondialdehyde (MDA) content and electrolyte leakage in cotton treated with VOCs were lower than those in the control group, and the expression levels of chitinase (CHI) and PR genes (PR10 and PR17), increased at various time points under the ST-TJ4 treatment. The activity of phenylalanine ammonia lyase (PAL) enzymes in cotton treated with VOCs was approximately 1.26 times greater than that in control plants at 24 h，while the contents of phenols and flavonoids increased significantly in the later stage. Additionally, 2-undecanone and 1-nonanol can induce a response in plants that enhances disease resistance. Collectively, these findings strongly suggest that VOCs from ST-TJ4 act as elicitors of plant defence and are valuable natural products for controlling Verticillium wilt.

Genome assembly of Hibiscus sabdariffa L. provides insights into metabolisms of medicinal natural products.

Hibiscus sabdariffa L. is a widely cultivated herbaceous plant with diverse applications in food, tea, fiber, and medicine. In this study, we present a high-quality genome assembly of H. sabdariffa using more than 33 Gb of high-fidelity (HiFi) long-read sequencing data, corresponding to ∼20× depth of the genome. We obtained 3 genome assemblies of H. sabdariffa: 1 primary and 2 partially haplotype-resolved genome assemblies. These genome assemblies exhibit N50 contig lengths of 26.25, 11.96, and 14.50 Mb, with genome coverage of 141.3, 86.0, and 88.6%, respectively. We also utilized 26 Gb of total RNA sequencing data to predict 154k, 79k, and 87k genes in the respective assemblies. The completeness of the primary genome assembly and its predicted genes was confirmed by the benchmarking universal single-copy ortholog analysis with a completeness rate of 99.3%. Based on our high-quality genomic resources, we constructed genetic networks for phenylpropanoid and flavonoid metabolism and identified candidate biosynthetic genes, which are responsible for producing key intermediates of roselle-specific medicinal natural products. Our comprehensive genomic and functional analysis opens avenues for further exploration and application of valuable natural products in H. sabdariffa.

Designing a highly efficient type III polyketide whole-cell catalyst with minimized byproduct formation

BackgroundPolyketide synthases (PKSs) are classified into three types based on their enzyme structures. Among them, type III PKSs, catalyzing the iterative condensation of malonyl-coenzyme A (CoA) with a CoA-linked starter molecule, are important synthases of valuable natural products. However, low efficiency and byproducts formation often limit their applications in recombinant overproduction.ResultsHerein, a rapid growth selection system is designed based on the accumulation and derepression of toxic acyl-CoA starter molecule intermediate products, which could be potentially applicable to most type III polyketides biosynthesis. This approach is validated by engineering both chalcone synthases (CHS) and host cell genome, to improve naringenin productions in Escherichia coli. From directed evolution of key enzyme CHS, beneficial mutant with ~ threefold improvement in capability of naringenin biosynthesis was selected and characterized. From directed genome evolution, effect of thioesterases on CHS catalysis is first discovered, expanding our understanding of byproduct formation mechanism in type III PKSs. Taken together, a whole-cell catalyst producing 1082 mg L−1 naringenin in flask with E value (evaluating product specificity) improved from 50.1% to 96.7% is obtained.ConclusionsThe growth selection system has greatly contributed to both enhanced activity and discovery of byproduct formation mechanism in CHS. This research provides new insights in the catalytic mechanisms of CHS and sheds light on engineering highly efficient heterologous bio-factories to produce naringenin, and potentially more high-value type III polyketides, with minimized byproducts formation.

An Unusual Ferryl Intermediate and Its Implications for the Mechanism of Oxacyclization by the Loline-Producing Iron(II)- and 2-Oxoglutarate-Dependent Oxygenase, LolO.

N-Acetylnorloline synthase (LolO) is one of several iron(II)- and 2-oxoglutarate-dependent (Fe/2OG) oxygenases that catalyze sequential reactions of different types in the biosynthesis of valuable natural products. LolO hydroxylates C2 of 1-exo-acetamidopyrrolizidine before coupling the C2-bonded oxygen to C7 to form the tricyclic loline core. Each reaction requires cleavage of a C-H bond by an oxoiron(IV) (ferryl) intermediate; however, different carbons are targeted, and the carbon radicals have different fates. Prior studies indicated that the substrate-cofactor disposition (SCD) controls the site of H· abstraction and can affect the reaction outcome. These indications led us to determine whether a change in SCD from the first to the second LolO reaction might contribute to the observed reactivity switch. Whereas the single ferryl complex in the C2 hydroxylation reaction was previously shown to have typical Mössbauer parameters, one of two ferryl complexes to accumulate during the oxacyclization reaction has the highest isomer shift seen to date for such a complex and abstracts H· from C7 ∼ 20 times faster than does the first ferryl complex in its previously reported off-pathway hydroxylation of C7. The detectable hydroxylation of C7 in competition with cyclization by the second ferryl complex is not enhanced in 2H2O solvent, suggesting that the C2 hydroxyl is deprotonated prior to C7-H cleavage. These observations are consistent with the coordination of the C2 oxygen to the ferryl complex, which may reorient its oxo ligand, the substrate, or both to positions more favorable for C7-H cleavage and oxacyclization.

Palladium-Catalyzed Amino-Sulfonylation of Aryl Iodide Derivatives via the Insertion of Sulfur Dioxide: One-Pot Synthesis of Aryl Primary Sulfonamides with Thiourea Dioxides

AbstractA palladium-catalyzed one-pot amino-sulfonylation of aryl iodide derivatives with thiourea dioxide, PdCl2dppf, and one-pot added hydroxylamine-O-sulfonic acid is presented. This amino-sulfonylation gave structure diversity to aryl primary sulfonamides and features good functional group compatibility, mild reaction conditions, excellent regioselectivity, and moderate to good yields. The robustness and potential of this method have also been successfully demonstrated by late-stage elaboration and gram-scale reaction. This approach achieves the divergent construction of the complex core structures that are prevalent in highly valuable natural products such as Sulpiride, Venetoclax, and Furosemide­.

Genomic insights into an endophytic Streptomyces sp. VITGV156 for antimicrobial compounds.

Endophytic Streptomyces sp. are recognized as a potential resource for valuable natural products but are less explored. This study focused on exploring endophytic Streptomyces species residing within tomato plants (Solanum lycopersicum) harboring genes for the production of a novel class of antibiotics. Our research involved the isolation and characterization of Streptomyces sp. VITGV156, a newly identified endophytic Streptomyces species that produces antimicrobial products. VITGV156 harbors a genome of 8.18 mb and codes 6,512 proteins, of which 4,993 are of known function (76.67%) and 1,519 are of unknown function (23.32%). By employing genomic analysis, we elucidate the genome landscape of this microbial strain and shed light on various BGCs responsible for producing polyketide antimicrobial compounds, with particular emphasis on the antibiotic kendomycin. We extended our study by evaluating the antibacterial properties of kendomycin. Overall, this study provides valuable insights into the genome of endophytic Streptomyces species, particularly Streptomyces sp. VITGV156, which are prolific producers of antimicrobial agents. These findings hold promise for further research and exploitation of pharmaceutical compounds, offering opportunities for the development of novel antimicrobial drugs.

Chemical Constituents and Bioactivities of Nepeta Taxa Essential Oils from Turkey: Principal Component Analysis, Molecular Docking Study, Molecular Dynamics, MM‐PBSA and Drug‐Likeness Estimation

AbstractNepeta taxa are popular plants and sources of traditional Turkish medicine. Chemical constituents in the essential oils (EOs) hydrodistilled from above‐ground parts of Nepeta aristata (NA), N. baytopii (NB), N. italica (NI), N. nuda (NN), and N. stenantha (NS) were determined using GC‐MS. The pharmacological activities of EOs, such as biological activities, were evaluated in vitro. DNA protection activities of NS‐EO were found to be the most effective. The NA‐EO, NI‐EO, and NS‐EO were more potent urease inhibitors, while the NS‐EO exhibited potent AChE inhibitors. Their molecular docking, molecular dynamics and enzyme inhibition results were compatible with the major components. Furthermore, MolDock and binding affinity results were found to be high. Moreover, AChE‐tau‐cadinol, BChE‐tau‐cadinol and tyrosinase‐Caryophyllene oxide complexes showed stability when simulated for 100 ns, with high MM‐PBSA energy values. In addition, the main components′ pharmacokinetic properties were observed to be effective. The present study showed that EOs may be valuable natural products for future pharmacological research, food, and the cosmetic industry according to in vitro and in silico values.

Enzyme-mediated ultrasound-assisted extraction of rutin, quercetin, and other antioxidant and antidiabetic compounds from Holarrhena antidysenterica seeds and optimization as per RSM

Rutin and quercetin are valuable bioactive natural products. The current study successfully discovered an efficient extraction protocol for these and other bioactive compounds from Holarrhena antidysenterica (HA) seeds based on an enzyme-mediated ultrasound-assisted extraction (EM-UAE). Extraction process optimization was done based on response surface methodology with seven response factors including total extraction yield (%), amount of rutin (AOR, mg/g), amount of quercetin (AOQ, mg/g), total phenolic content (TPC, mg gallic acid equivalents/g dry weight (mg GAE/g DW), total flavonoids content (TFC, mg rutin equivalents/g DW (mg RE/g DW) anti-radical activity (ARA, %), and α-amylase inhibitory activity (AIA, %). High-performance liquid chromatography (HPLC) was used for quantification of rutin and quercetin. The optimal extraction yield, AOR, AOQ, TPC, TFC, ARA, and AIA were 32.39 %, 71.43 mg/g, 23.68 mg/g, 178.63 mg GAE/g DW, 100.22 mg RE/g DW, 76.41 %, and 77.48 %, respectively. The optimal conditions were 91 min time, 6.1 mL enzyme cocktail, pH 4.8, and 48 ℃ temperature. For ARA, EC50 (µg/mL) of HA was 9.75 ± 0.27 compared to 6.10 ± 0.38 of ascorbic acid standard. For AIA, IC50 (µg/mL) of HA and acarbose standard were 51.97 ± 0.21 and 47.89 ± 0.61 μg/mL, respectively. The validation study strongly supported the predicted model with a very low relative standard deviation (0.45–3.37 %). The model was statistically significant and efficient for obtaining rutin, quercetin, and other antioxidant and antidiabetic compounds from Holarrhena antidysenterica seeds on the industrial level.

Research progress on the pharmacological activity, biosynthetic pathways, and biosynthesis of crocins.

Crocins are water-soluble apocarotenoids isolated from the flowers of crocus and gardenia. They exhibit various pharmacological effects, including neuroprotection, anti-inflammatory properties, hepatorenal protection, and anticancer activity. They are often used as coloring and seasoning agents. Due to the limited content of crocins in plants and the high cost of chemical synthesis, the supply of crocins is insufficient to meet current demand. The biosynthetic pathways for crocins have been elucidated to date, which allows the heterologous production of these valuable compounds in microorganisms by fermentation. This review article provides a comprehensive overview of the chemistry, pharmacological activity, biosynthetic pathways, and heterologous production of crocins, aiming to lay the foundation for the large-scale production of these valuable natural products by using engineered microbial cell factories.

Engineering the activity and thermostability of a carboxylic acid reductase in the conversion of vanillic acid to vanillin

Vanillin is a valuable natural product that can be used as a fragrance and additive. Recent research in the biosynthesis of vanillin has brought attention to a key enzyme, carboxylic acid reductase (CAR), which catalyzes the reduction of vanillic acid to vanillin. Nevertheless, the biosynthesis of vanillin is hampered by the low activity and stability of CAR. As such, a rational design campaign was conducted on a well-documented carboxylic acid reductase from Segniliparus rugosus (SrCAR), using vanillic acid as the model substrate. After combined active site saturation and iterative site-specific mutagenesis, the best quadruple mutant N292H/K524S/A627L/E1121W (M3) was successfully obtained. In comparison to the wildtype SrCAR, M3 demonstrated a 4.2-fold increase in catalytic efficiency (kcat/Km), and its half-life (t1/2) was enhanced by 3.8 times up to 385.08 minutes at 40 °C. In silico docking and molecular dynamics simulation provided insights into the improved activity and stability. In the subsequent preparative-scale reaction with 100 mM (16.8 g L−1) vanillic acid, the whole cell catalysis utilizing M3 produced 10.15 g·L−1 of vanillin and 1.11 g·L−1 of vanillyl alcohol, respectively. This work demonstrates a dual improvement in the activity and thermal stability of SrCAR, thereby potentially facilitating the application of carboxylic acid reductase in the biosynthesis of vanillin.

Space-Efficient 3D Microalgae Farming with Optimized Resource Utilization for Regenerative Food.

Photosynthetic microalgae produce valuable metabolites and are a source of sustainable food that supports life without compromising arable land. However, the light self-shading, excessive water supply, and insufficient space utilization in microalgae farming have limited its potential in the inland areas most in need of regenerative food solutions. Herein, this work develops a 3D polysaccharide-based hydrogel scaffold for vertically farming microalgae without needing liquid media. This liquid-free strategy is compatible with diverse microalgal species and enables the design of living microalgal frameworks with customizable architectures that enhance light and water utilization. This approach significantly increases microalgae yield per unit water consumption, with an 8.8-fold increase compared to traditional methods. Furthermore, the dehydrated hydrogels demonstrate a reduced size and weight (≈70% reduction), but readily recover their vitality upon rehydration. Importantly, valuable natural products can be produced in this system including proteins, carbohydrates, lipids, and carotenoids. This study streamlines microalgae regenerative farming for low-carbon biomanufacturing by minimizing light self-shading, relieving water supply, and reducing physical footprints, and democratizing access to efficient aquatic food production.