Metabolic engineering最新文献

{"title":"Multisite Cre-lox recombination enables regulatory mechanism elucidation and systematic engineering of echinocandin B biosynthesis in Aspergillus nidulans","authors":"Youchu Ma ,&nbsp;Yue Zhang ,&nbsp;Dongfang Zhang ,&nbsp;Yue Tang ,&nbsp;Wanyu Zhang ,&nbsp;Liyuan Yue ,&nbsp;Wenqing Lou ,&nbsp;Chao Meng ,&nbsp;Yanling Li ,&nbsp;Xiulai Chen ,&nbsp;Fanglong Zhao","doi":"10.1016/j.ymben.2026.01.009","DOIUrl":"10.1016/j.ymben.2026.01.009","url":null,"abstract":"<div><div>Echinocandin B (ECB), a cyclic lipohexapeptide for synthesizing antifungal drugs, is produced by the nonribosomal peptide synthetase gene cluster in <em>Aspergillus nidulans</em>. However, industrial production remains limited by the inefficiency of production capacity, primarily due to the complexity of the biosynthetic pathway and the absence of multi-gene regulatory tools in filamentous fungi. Here, we established an orthogonal Cre-<em>lox</em>-based platform enabling single-site insertion of up to 30 kb and simultaneous dual-site integration of 10 kb DNA fragments in <em>A. nidulans</em>. Through precursor supplementation and targeted gene overexpression, we identified key enzymatic bottlenecks in the precursor biosynthetic pathway, including the oxygenases AniF<em>,</em> AniK<em>,</em> AniG, and the acyl-AMP ligase AniI. Combinatorial overexpression of these genes acted synergistically to increase ECB titers. We further addressed bottlenecks in natural amino acid biosynthesis by overexpressing feedback-resistant mutants of Hom3 (L-Thr pathway) and LeuC (L-Leu pathway). Additionally, we uncovered a temperature-dependent regulation mechanism whereby low temperature (25 °C) concurrently upregulates both the ECB biosynthetic gene cluster and <em>odeA</em> gene, encoding <em>Δ</em>12-oleic acid desaturase, thereby increasing linoleic acid availability for ECB production. Leveraging our multisite DNA-integration platform to rewire expression of these key genes, we increased ECB production to 3.5 ± 0.2 g/L in a 5-L fed-batch bioreactor, a 2.3-fold improvement that represents the highest titer reported in the literature to date. Our orthogonal dual-site integration strategy and the systematic optimization approach provide a valuable framework for metabolic engineering of complex natural products in filamentous fungi.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"95 ","pages":"Pages 15-27"},"PeriodicalIF":6.8,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146033201","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Investigating overflow metabolism in heterotrophic cultures of the green alga Chromochloris zofingiensis","authors":"Michelle Meagher ,&nbsp;Dimitrios J. Camacho ,&nbsp;Sean D. Gallaher ,&nbsp;Sabeeha S. Merchant ,&nbsp;Nanette R. Boyle","doi":"10.1016/j.ymben.2026.01.007","DOIUrl":"10.1016/j.ymben.2026.01.007","url":null,"abstract":"<div><div><em>Chromochloris zofingiensis</em> is of interest for its ability to perform a reversible trophic switch in the presence of glucose that is characterized by a shutdown of photosynthesis and an accumulation of energy storage metabolites. Previous work has shown that this trophic switch is accompanied by overflow metabolism and the production of lactate in aerobic conditions. This trophic switch is not observed in nutrient replete media. We utilized isotopically assisted metabolic flux analysis to characterize intracellular flux distributions that are associated with different metabolic phenotypes observed in this organism in different media formulations in light and dark conditions. The results of this analysis showed that low iron cultures have no flux through carbon fixation reactions, and that the carbon flux entering the TCA cycle in these cultures is approximately 40 % lower than that in iron replete cultures grown heterotrophically. This analysis was complemented with transcriptomics data collected for <em>C. zofingiensis</em> grown in iron limited conditions to provide further evidence towards the negative impact of iron limitation on both photosynthetic and respiratory activity. Overflow metabolism allows this alga to compensate for the lower energy production that results from iron limitation. This work highlights how nutrient availability can lead to changes in the metabolism of <em>C. zofingiensis</em>.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"95 ","pages":"Pages 1-14"},"PeriodicalIF":6.8,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145995162","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Reconstruction of arginine deiminase pathway sustains a higher-energy state in mammalian cells","authors":"Mauro Torres ,&nbsp;Matthew Reaney ,&nbsp;Kate Meeson ,&nbsp;Devika Kalsi ,&nbsp;Leon P. Pybus ,&nbsp;Alan J. Dickson","doi":"10.1016/j.ymben.2026.02.004","DOIUrl":"10.1016/j.ymben.2026.02.004","url":null,"abstract":"<div><div>ATP is the universal “energy currency” of the cell, and its supply may represent one of several limiting factors influencing the productivity of mammalian cell factories. Here, we present a novel, mitochondria-independent approach to enhance cellular energy metabolism. We engineered Chinese hamster ovary (CHO) cells to express the bacterial arginine deiminase (ADI) pathway along with two arginine transporters. This system enables the direct, cytosolic conversion of arginine to ATP, effectively generating energy without relying solely on the cell's native metabolic machinery. ADI pathway expression was associated with intracellular ATP and concurrent improvements in culture performance across different CHO cell backgrounds. In contrast, cell lines engineered only for enhanced arginine uptake showed no performance gain, consistent with the hypothesis that de-novo ATP generation may contribute to improve productivity. Metabolic profiling revealed that the ADI pathway affects cellular metabolism. We observed a downshift in glycolysis, characterized by decreased glucose consumption and reduced lactate and alanine production, while amino acid and TCA cycle intermediaries remained broadly unchanged. Adenylate measurements and AMPK signalling analysis confirmed a higher energy state (ATP↑, ADP/ATP↓, p-AMPK/AMPK↓) in engineered cells. Supplementing the cell culture medium with arginine or citrulline was associated with further increases in growth and mAb titres in ADI-expressing cells. These results establish the ADI pathway as a powerful and distinct method for enhancing cellular energy. This mitochondria-independent approach highlights a new paradigm for improving the efficiency of industrial bioprocesses.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"95 ","pages":"Pages 63-76"},"PeriodicalIF":6.8,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146153271","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metabolic engineering of Escherichia coli for the high-level production of putrescine","authors":"Yoo-Sung Ko ,&nbsp;Je Woong Kim ,&nbsp;Alisher Nazarbekov ,&nbsp;Gi Bae Kim ,&nbsp;Sang Yup Lee","doi":"10.1016/j.ymben.2026.01.006","DOIUrl":"10.1016/j.ymben.2026.01.006","url":null,"abstract":"<div><div>Putrescine is an important platform chemical used in the manufacture of engineering plastics. To support the development of a sustainable plastics industry, microbial production of putrescine from renewable resources has attracted increasing attention. In this study, we report the development of an engineered <em>Escherichia coli</em> strain capable of efficiently producing putrescine. To overcome the limitation caused by putrescine toxicity, the previously developed XQ52 strain (a W3110-derived putrescine-producing strain) was subjected to adaptive laboratory evolution, resulting in the AXQ52 strain, which produced 61.7 g/L of putrescine in fed-batch fermentation. This titer surpassed the natural tolerance threshold of <em>E. coli</em>. Genome sequencing of the AXQ52 strain revealed mutations that improved cellular fitness under high putrescine concentrations. Further production improvements were achieved by fine-tuning the expression of phosphoenolpyruvate carboxylase gene, introducing a heterologous ornithine acetyltransferase, and disrupting glutamate decarboxylase. The final engineered strain produced 72.7 g/L of putrescine with a yield of 0.25 g/g glucose and a productivity of 1.28 g/L/h, representing the highest microbial putrescine production reported to date from a simple carbon source.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"95 ","pages":"Pages 28-38"},"PeriodicalIF":6.8,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145995006","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Omics analyses decoding mechanisms underlying the self-flocculating phenotype of yeast cells and stress tolerance for robust production","authors":"Xue Zhang ,&nbsp;Yang Dai ,&nbsp;Xin-Qing Zhao ,&nbsp;Chen-Guang Liu ,&nbsp;Zhuo Wang ,&nbsp;Feng-Wu Bai","doi":"10.1016/j.ymben.2026.01.003","DOIUrl":"10.1016/j.ymben.2026.01.003","url":null,"abstract":"<div><div>A unique self-flocculating yeast strain SPSC01 was developed through protoplast fusion for fuel ethanol production with high product titers. In this study, we conducted comparative multi-omics analyses on SPSC01 to elucidate mechanisms underlying its self-flocculating phenotype and associated stress tolerance, the most desirable merit for robust production in industry. Leveraging two cutting-edge third-generation sequencing technologies, we achieved a gapless high-quality and chromosome-level assembly for the genomes of SPSC01 and its parental strains. Through comprehensive genome analyses, we identified 25 unique genes that are absent in the parental strains, along with 13 novel genes with unknown functions. The self-flocculation of yeast cells is driven by the copy number of genetic variations and significantly upregulated transcription of <em>FLO</em> genes. Mutations in both <em>cis</em>- and <em>trans</em>-regulatory elements contribute to the constitutive expression of <em>FLO1</em> and its derivative genes, a prerequisite for developing the self-flocculating phenotype. Notably, we discovered a novel small protein G12 that harbors a zinc finger domain, and its overexpression substantially enhanced ethanol production of engineered yeast strains. Furthermore, alterations in metabolic pathways with ergosterol, glutathione, amino acid, and glycerophospholipid are implicated for developing tolerance to ethanol and major inhibitors acetic acid and furfural that are released during the pretreatment of lignocellulosic biomass. The progress provides strategies for engineering yeast cell factories with robustness through rational design to produce biofuels and bio-based chemicals with high product titers and productivities, in particular with the biorefinery of lignocellulosic biomass for sustainable socioeconomic development.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"95 ","pages":"Pages 77-89"},"PeriodicalIF":6.8,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145937781","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineering Halomonas bluephagenesis for high-efficiency biosynthesis of pyruvate","authors":"Kang Wang ,&nbsp;Zonghao Zhang ,&nbsp;Zhongnan Zhang ,&nbsp;Fuqing Wu ,&nbsp;Guo-Qiang Chen","doi":"10.1016/j.ymben.2026.02.003","DOIUrl":"10.1016/j.ymben.2026.02.003","url":null,"abstract":"<div><div>Pyruvate, a C3 platform compound, has significant applications across multiple sectors, including bio-based materials (e.g., polylactic acid), pharmaceutical intermediates (such as L-alanine), and food additives. Biomanufacturing via microbial fermentation provides renewable feedstocks and cleaner processes compared to traditional petroleum-based methods. This study explores the extremophilic halophile <em>Halomonas bluephagenesis</em> TD01 as a chassis organism for pyruvate production. To enhance pyruvate synthesis, several engineering strategies were implemented, including blocking the primary carbon consumption pathway, eliminating pyruvate bypass degradation, reducing tricarboxylic acid cycle activity, removing the glycolic acid cycle, regulating transcription factors, and minimizing pyruvate reabsorption and utilization. The engineered <em>H. bluephagenesis</em> TD1.24 produced 39 g/L pyruvate in a 50-h non-sterile fed-batch fermentation. Simultaneously, the high-pyruvate-producing strains showed improved conversion rate of PHB and efficient acetoin synthesis. <em>H. bluephagenesis</em> demonstrated robustness as a chassis for next generation industrial biotechnology (NGIB), enabling the production of both its native and a broader range of biological products.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"95 ","pages":"Pages 50-62"},"PeriodicalIF":6.8,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146138368","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineering xylose catabolism in the yeast Komagataella phaffii","authors":"Kun Zhang ,&nbsp;Xin Ni ,&nbsp;Peng Cai ,&nbsp;Yongjin J. Zhou","doi":"10.1016/j.ymben.2026.02.001","DOIUrl":"10.1016/j.ymben.2026.02.001","url":null,"abstract":"<div><div>Efficient xylose utilization is crucial for biomass hydrolysate valorization. However, <em>Komagataella phaffii</em> cannot efficiently utilize xylose. Here, we constructed xylose isomerase (XI)-xylulokinase (XK) pathway, nonoxidative pentose phosphate pathway (PPP), and nonoxidative glycolysis (NOG) pathway in <em>K. phaffii</em> to increase cell growth on xylose. Additionally, the bypass pathway of xylose metabolism, the high osmolarity glycerol/mitogen-activated protein kinase (HOG-MAPK) signaling pathway, and possible negative transcription factors were blocked to further promote xylose catabolism. Moreover, adaptive laboratory evolution (ALE) dramatically improved xylose utilization, and six potential targets were identified through multiomics and reverse engineering. The engineered strain exhibited the highest reported specific growth rate μ_max of up to 0.042 h⁻¹ and lag time of 25.0 h with a biomass yield of 0.366 g dry cell weight/g from sole xylose in minimal media. This strain also showed faster metabolite turnover, efficient free fatty acid (FFA) production and partial amelioration of glucose repression effect from xylose alone. The engineered metabolic plasticity described here will facilitate the regulation of xylose catabolism in other nonnative xylose-consuming yeasts.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"95 ","pages":"Pages 39-49"},"PeriodicalIF":6.8,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146134407","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineering amino acid-derived malonyl-CoA pathways to boost polyketide production in Yarrowia lipolytica","authors":"Jinpeng Wang ,&nbsp;Yuxiang Hong ,&nbsp;Zizhao Wu ,&nbsp;Ayelet Fishman ,&nbsp;Peng Xu","doi":"10.1016/j.ymben.2025.11.015","DOIUrl":"10.1016/j.ymben.2025.11.015","url":null,"abstract":"<div><div>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 pathways leverage oxidative deamination and decarboxylation reactions and efficiently channel metabolic flux toward malonyl-CoA, resulting in increased production of total polyketides beyond the capacity of the native acetyl-CoA carboxylase route using glucose as substrate. We also discovered a new-to-nature polyketide (4-hydroxy-6-hydroxyethyl-2-pyrone) derived from the side activity of the TAL pathway, reaching 6.4 g/L in <em>Y. lipolytica</em>. This work highlights the utility of the novel malonyl-CoA pathways in enhancing polyketide production, and the possibility of upcycling abundant amino acids or protein waste in the animal farming or meat industry to produce high-value nonnatural polyketides.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"94 ","pages":"Pages 99-109"},"PeriodicalIF":6.8,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145553880","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Disruption of acyl-acyl carrier protein (acyl-ACP) synthetase in cyanobacteria impairs lipid remodeling as revealed by acyl-ACP measurements","authors":"Juthamas Jaroensuk ,&nbsp;Joshua P. Abraham ,&nbsp;Baltazar E. Zuniga ,&nbsp;Hawkins S. Shepard ,&nbsp;Michael Wei ,&nbsp;Russell Williams ,&nbsp;Stewart A. Morley ,&nbsp;Maneesh Lingwan ,&nbsp;Jiahong Zhou ,&nbsp;Michael A. Jindra ,&nbsp;Poonam Jyoti ,&nbsp;Bo Wang ,&nbsp;Jody C. May ,&nbsp;John A. McLean ,&nbsp;Jamey D. Young ,&nbsp;Brian F. Pfleger ,&nbsp;Doug K. Allen","doi":"10.1016/j.ymben.2025.11.004","DOIUrl":"10.1016/j.ymben.2025.11.004","url":null,"abstract":"<div><div>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, temperature shifts are known to affect fatty acid desaturation and membrane fluidity, yet it is unclear whether AAS contributes to these adaptive responses through re-esterification of membrane-released acyl chains. We elucidated unique aspects of fatty acid metabolism in response to temperature changes in biotechnologically relevant microbes with the development of an efficient method for quantifying acyl-ACP intermediates using anion exchange chromatography (AEX). In <em>Escherichia coli,</em> which performs desaturation during fatty acid biosynthesis, we detected saturated and unsaturated acyl-ACPs that confirm biosynthetic pathway operation. In the cyanobacteria, <em>Picosynechococcus</em> sp. PCC 7002 and the Δ<em>aas</em> strain, changes between two temperatures were interpreted with support from proteomic and lipidomic analyses and indicated that the AAS is tied to membrane lipid remodeling. Further, polyunsaturated acyl-ACPs were detected in the Δ<em>aas</em> strain, which was unexpected because fatty acid synthesis does not produce polyunsaturates in cyanobacteria, suggesting the presence of alternative acyl-activating enzymes or unknown acyl-ACP desaturases. This study highlights the possible link between acyl chain recycling and lipid remodeling in cyanobacteria and demonstrates the utility of AEX-based acyl-ACP profiling in dissecting fatty acid metabolism.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"94 ","pages":"Pages 45-56"},"PeriodicalIF":6.8,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145447241","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineered plants for the production of the antioxidants arbutin and gallate","authors":"Sami Kazaz ,&nbsp;Yu-Ton Chen ,&nbsp;Senri Yamamoto ,&nbsp;Yang Tian ,&nbsp;Chien-Yuan Lin ,&nbsp;Dylan Chin ,&nbsp;İrem Pamukçu ,&nbsp;Ibraheem Mohammed Al Shammaa ,&nbsp;Yusuf Selman Akbas ,&nbsp;Monikaben Nimavat ,&nbsp;Emine Akyuz Turumtay ,&nbsp;Edward E.K. Baidoo ,&nbsp;Albert P. Kausch ,&nbsp;Yuki Tobimatsu ,&nbsp;Aymerick Eudes","doi":"10.1016/j.ymben.2025.11.009","DOIUrl":"10.1016/j.ymben.2025.11.009","url":null,"abstract":"<div><div>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 in <em>Nicotiana benthamiana</em> leaves, we first identified biosynthetic routes for the production of gallate from either chorismate or 3-dehydroshikimate. Gallate production was then achieved in Arabidopsis using a genetic background that overproduces protocatechuate and via expression of a mutated version of the 4-hydroxybenzoate hydroxylase PobA from <em>Pseudomonas</em> sp. Arbutin production was obtained in Arabidopsis using a genetic background that overproduces 4-hydroxybenzoate and via expression of the monooxygenase MNX1 from <em>Candida parapsilosis</em>. The best Arabidopsis transgenic lines accumulated gallate and arbutin in the range of 0.25 and 0.93 dry weight % (dwt%), respectively. Using sorghum for large-scale <em>in planta</em> production, the titers of gallate and arbutin produced from the intermediate 4-hydroxybenzoate reached 0.58 dwt% and 0.50 dwt%, respectively, in mature transgenic plants, surpassing levels typically observed in plants that naturally produce these compounds. Gallate and arbutin were readily extracted from plant tissues using methanol solvent. Analysis of extractive-free biomass showed only trace amounts of gallate and its precursors 4-hydroxybenzoate and protocatechuate crosslinked to cell walls, suggesting that they mainly occur as soluble conjugated forms stored in the vacuole. This study presents alternative synthesis routes using plant hosts for the eco-friendly production of gallate and arbutin.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"94 ","pages":"Pages 57-66"},"PeriodicalIF":6.8,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145492048","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}