With the progress of urbanization,rural tourism has emerged as a popular leisure activity in China.The crop field with pattern art has been gaining popularity over the years,using strains with colorful leaves to creat...With the progress of urbanization,rural tourism has emerged as a popular leisure activity in China.The crop field with pattern art has been gaining popularity over the years,using strains with colorful leaves to create various impressive designs(Xu 2024).A lot of successful cases in rice fields have not only attracted tourists but also increased the income of farmers(Song et al.2020).Plants with colorful leaves also have gained significant popularity in ornamental agriculture.Therefore,developing plants with colorful leaves has gained significant popularity in ornamental agriculture.展开更多
To improve the nutritional and functional value of rice,numerous biotechnological approaches have focused on metabolic engineering to address nutritional deficiencies and produce health-beneficial compounds that are e...To improve the nutritional and functional value of rice,numerous biotechnological approaches have focused on metabolic engineering to address nutritional deficiencies and produce health-beneficial compounds that are either absent or naturally present in low amounts.A prominent example is‘Golden Rice’,which has been genetically modified to accumulateβ-carotene to combat vitamin A deficiency in regions with limited dietary intake.Scientists have been continuously biofortifying rice with various specialized metabolites,including terpenoids,flavonoids,non-flavonoid polyphenols,betalains,vitamins,and amino acids.This review explores the specific pathways and genetic modifications utilized by researchers to enhance the accumulation of targeted metabolites in rice.It comprehensively summarizes key strategies and research trends in rice metabolic engineering,demonstrating how rice can be transformed into a strategic crop for producing industrially valuable compounds beyond its traditional role as a staple food by leveraging its advantages as a versatile host system through its grains,leaves,and cells.Furthermore,we highlight the potential of intergrating metabolic engineering with synthetic biology and big data-driven computational modeling,particularly through artificial intelligence and machine learning,as promising future research directions.展开更多
Tyrosol is a natural phenolic compound with antioxidant,anti-inflammatory and other biological activities,serving as an important precursor of high-value products such as hydroxytyrosol and salidroside.Therefore,the g...Tyrosol is a natural phenolic compound with antioxidant,anti-inflammatory and other biological activities,serving as an important precursor of high-value products such as hydroxytyrosol and salidroside.Therefore,the green and efficient biosynthesis of tyrosol and its derivatives has become a research hotspot in recent years.Building cell factories by metabolic engineering of microorganisms is a potential industrial production way,which has low costs and environmental friendliness.This paper introduces the biosynthesis pathway of tyrosol and presents the key regulated nodes in the de novo synthesis of tyrosol in Escherichia coli and Saccharomyces cerevisiae.In addition,this paper reviews the recent advances in metabolic engineering for the production of hydroxytyrosol and salidroside.This review can provide a reference for engineering the strains for the high-yield production of tyrosol and its derivatives.展开更多
Metabolic engineering of the tumor microenvironment has emerged as a new strategy.Lactate dehydrogenase A(LDHA)is a prominent target for metabolic engineering.Here,we designed a cationic lipid nanoparticle formulation...Metabolic engineering of the tumor microenvironment has emerged as a new strategy.Lactate dehydrogenase A(LDHA)is a prominent target for metabolic engineering.Here,we designed a cationic lipid nanoparticle formulation for LDHA gene editing.The plasmid DNA delivery efficiency of our lipid nanoparticle formulations was screened by testing the fluorescence of lipid nanoparticles complexed to plasmid DNA encoding green fluorescence protein(GFP).The delivery efficiency was affected by the ratios of three components:a cationic lipid,cholesterol or its derivative,and a fusogenic lipid.The lipid nanoparticle designated formulation F3 was complexed to plasmid DNA co-encoding CRISPR-associated protein 9 and LDHA-specific sgRNA,yielding the lipoplex,pCas9-sgLDHA/F3.The lipoplex including GFP-encoding plasmid DNA provided gene editing in HeLa-GFP cells.Treatment of B16F10 tumor cells with pCas9-sgLDHA/F3 yielded editing of the LDHA gene and increased the pH of the culture medium.pCas9-sgLDHA/F3 treatment activated the interferon-gamma and granzyme production of T cells in culture.In vivo,combining pCas9-sgLDHA/F3 with immune checkpoint-inhibiting anti-PD-L1 antibody provided a synergistic antitumor effect and prolonged the survival of tumor model mice.This study suggests that combining metabolic engineering of the tumor microenvironment with immune checkpoint inhibition could be a valuable antitumor strategy.展开更多
In recent years,metabolic engineering has made great progress in both academic research and industrial applications.However,we have not found any articles that specifically analyze the current state of metabolic engin...In recent years,metabolic engineering has made great progress in both academic research and industrial applications.However,we have not found any articles that specifically analyze the current state of metabolic engineering in China in comparison with other countries.Here,we review the current development and future trends of global metabolic engineering,conduct an in-depth benchmarking analysis of the development situation of China’s metabolic engineering,and identify current problems as well as future trends.We searched publications in the Scopus database from 2015 to September 2020 in the field of metabolic engineering,and analyzed the output in general,including publication trends,research distribution,popular journals,hot topics and vital institutions,but also analyzed the share of citations,field-weighted citation impact,and production in collaboration with strategic countries in science and technology.This study aims to serve as a reference for later studies,offering a comprehensive view of China’s contribution to metabolic engineering,and as a tool for the elaboration of national public policy in science and technology.展开更多
Through several waves of technological research and un‐matched innovation strategies,bio‐catalysis has been widely used at the industrial level.Because of the value of enzymes,methods for producing value‐added comp...Through several waves of technological research and un‐matched innovation strategies,bio‐catalysis has been widely used at the industrial level.Because of the value of enzymes,methods for producing value‐added compounds and industrially‐relevant fine chemicals through biological methods have been developed.A broad spectrum of numerous biochemical pathways is catalyzed by enzymes,including enzymes that have not been identified.However,low catalytic efficacy,low stability,inhibition by non‐cognate substrates,and intolerance to the harsh reaction conditions required for some chemical processes are considered as major limitations in applied bio‐catalysis.Thus,the development of green catalysts with multi‐catalytic features along with higher efficacy and induced stability are important for bio‐catalysis.Implementation of computational science with metabolic engineering,synthetic biology,and machine learning routes offers novel alternatives for engineering novel catalysts.Here,we describe the role of synthetic biology and metabolic engineering in catalysis.Machine learning algorithms for catalysis and the choice of an algorithm for predicting protein‐ligand interactions are discussed.The importance of molecular docking in predicting binding and catalytic functions is reviewed.Finally,we describe future challenges and perspectives.展开更多
Mushrooms are abundant in bioactive natural compounds.Due to strict growth conditions and long fermentation-time,microbe as a production host is an alternative and sustainable approach for the production of natural co...Mushrooms are abundant in bioactive natural compounds.Due to strict growth conditions and long fermentation-time,microbe as a production host is an alternative and sustainable approach for the production of natural compounds.This review focuses on the biosynthetic pathways of mushroom originated natural compounds and microbes as the production host for the production of the above natural compounds.展开更多
In animals,serotonin is a neurotransmitter and mood regulator.In plants,serotonin functions in energy acquisition,tissue maintenance,delay of senescence,and response to biotic and abiotic stresses.In this study,we exa...In animals,serotonin is a neurotransmitter and mood regulator.In plants,serotonin functions in energy acquisition,tissue maintenance,delay of senescence,and response to biotic and abiotic stresses.In this study,we examined the effect of serotonin enrichment of rice endosperm on plant growth,endosperm development,and grain quality.To do so,TDCs and T5H were selected as targets for serotonin fortification.Overexpression of TDC1 or TDC3 increased serotonin accumulation relative to overexpression of T5H in rice grain.Transgenic lines of target genes driven by the Gt1 promoter showed better field performance than those driven by the Ubi promoter.Overexpression of T5H showed little effect on plant growth or grain physicochemical quality.In neuronal cell culture assays,serotonin induced neuroprotective action against apoptosis.Breeding of rice cultivars with high serotonin content may be beneficial for health and nutrition.展开更多
Depleting global petroleum reserves and skyrocketing prices coupled with succinct supply have been a grave concern,which needs alternative sources to conventional fuels.Oleaginous microalgae have been explored for enh...Depleting global petroleum reserves and skyrocketing prices coupled with succinct supply have been a grave concern,which needs alternative sources to conventional fuels.Oleaginous microalgae have been explored for enhanced lipid production,leading towards biodiesel production.These microalgae have short life cycles,require less labor,and space,and are easy to scale up.Triacylglycerol,the primary source of lipids needed to produce biodiesel,is accumulated by most microalgae.The article focuses on different types of oleaginous microalgae,which can be used as a feedstock to produce biodiesel.Lipid biosynthesis in microalgae occurs through fatty acid synthesis and TAG synthesis approaches.In-depth discussions are held regarding other efficient methods for enhancing fatty acid and TAG synthesis,regulating TAG biosynthesis bypass methods,blocking competing pathways,multigene approach,and genome editing.The most potential targets for gene transformation are hypothesized to be a malic enzyme and diacylglycerol acyltransferase while lowering phosphoenolpyruvate carboxylase activity is reported to be advantageous for lipid synthesis.展开更多
Mesaconic acid has a special chemical structure and can undergo a series of reactions such as polymerization and addition. It is an important chemical intermediate and widely used in material, chemical and other indus...Mesaconic acid has a special chemical structure and can undergo a series of reactions such as polymerization and addition. It is an important chemical intermediate and widely used in material, chemical and other industries. The chemical synthesis of mesaconic acid requires nitric acid, which is dangerous and harmful to the environment. The production of mesaconic acid by microbial fermentation has the characteristics of low raw material price, high efficiency and strong specificity, and thus a strong industrial application prospect. Mesaconic acid is an intermediate product of glutamic acid degradation pathway of microorganisms such as Clostridium tetani. However, at present, few reports have been conducted on the production of mesaconic acid by metabolic engineering microorganisms. In this study, glutamate mutase(GLM) and 3-methylaspartate ammonialyase(MAL) from C. tetani were recombined and expressed in Escherichia coli, and the obtained strain, BL21(DE3)/pETDuet-1-MAL-mutS-mutE, achieved the yield of mesaconic acid of 1.06 g/L. Compared with the wild type, the yields of mesaconic acid from mutants G133A and G133S increased by 21% and 16%, respectively. After 24 h of flask fermentation, the yields of mesaconic acid reached 1.28 and 1.23 g/L, respectively. This study can provide reference for microbial synthesis of mesaconic acid.展开更多
Biosynthesis of paclitaxel(Taxol™)is a hot topic with extensive and durable interests for decades.However,it is severely hindered due to the very low titers of intermediates.In this study,Escherichia coli was employed...Biosynthesis of paclitaxel(Taxol™)is a hot topic with extensive and durable interests for decades.However,it is severely hindered due to the very low titers of intermediates.In this study,Escherichia coli was employed to de novo synthesize a key intermediate of paclitaxel,taxadien-5α-yl-acetate(T5OAc).Plasmid-based pathway reconstruction and optimization were conducted for T5OAc production.The endogenous methylerythritol phosphate pathway was enhanced to increase the precursor supply.Three taxadien-5α-ol O-acetyltransferases were tested to obtain the best enzyme for the acetylation step.Metabolic burden was relieved to restore cell growth and promote production through optimizing the plasmid production system.In order to achieve metabolic balance,the biosynthesis pathway was regulated precisely by multivariate-modular metabolic engineering.Finally,in a 5-L bioreactor,the T5OAc titer was enhanced to reach 10.9 mg/L.This represents an approximately 272-fold increase in production compared to the original strain,marking the highest yield of T5OAc ever documented in E.coli,which is believed to be helpful for promoting the progress of paclitaxel biosynthesis.展开更多
Objective:The biosynthesis of pilose antler polysaccharides(PAPS)with anti-aging activity relies on the precise regulation of key enzymes;however,the identification of these enzymes is limited by traditional low-throu...Objective:The biosynthesis of pilose antler polysaccharides(PAPS)with anti-aging activity relies on the precise regulation of key enzymes;however,the identification of these enzymes is limited by traditional low-throughput techniques.This study constructed an integrated system combining metabolic engineering and high-throughput screening of fluorescent substrates,aiming to overcome the technical bottlenecks in the identification of key enzymes for PAPS synthesis and the optimization of their catalytic efficiency.Methods:Eighteen candidate genes related to carbohydrate metabolism were screened via transcriptome sequencing of deer antler stem cells.After prokaryotic expression and preliminary screening by high-performance liquid chromatography(HPLC),a self-designed fluorescent substrate(FAM-Glc-β1-3-PNPG)combined with a 96-well plate platform was used to achieve high-throughput optimization and screening of catalytic efficiency.Enzyme function was verified through kinetic analysis and in vitro antioxidant experiments.Results:The screened high-efficiency glycosyltransferase UGT-Wnt3a showed a 2.1-fold increase in catalytic efficiency compared with the wild type,with a maximum reaction rate(Vmax)of12.3μmol·min^(-1)·mg^(-1)and a Michaelis constant(Km)of 0.87 mM.The catalytic product of UGT-Wnt3a increased the superoxide dismutase(SOD)activity by 42%(P<0.01)and decreased the malondialdehyde(MDA)content by 28%(P<0.05)in the oxidative damage system.The detection efficiency of the new platform was 9 times higher than that of HPLC,and the limit of detection was reduced by 50 times.Conclusion:The screening system established in this study provides an innovative technical solution for the directed synthesis of PAPS and the development of anti-aging components,promoting the transformation of active components in traditional Chinese medicine from natural extraction to bioengineering-based production.展开更多
Vitamin A is a micronutrient critical for versatile biological functions and has been widely used in the food,cosmetics,pharmaceutical,and nutraceutical industries.Synthetic biology and metabolic engineering enable mi...Vitamin A is a micronutrient critical for versatile biological functions and has been widely used in the food,cosmetics,pharmaceutical,and nutraceutical industries.Synthetic biology and metabolic engineering enable microbes,especially the model organism Saccharomyces cerevisiae(generally recognised as safe)to possess great potential for the production of vitamin A.Herein,we first generated a vitamin A-producing strain by miningβ-carotene 15,15′-mono(di)oxygenase from different sources and identified two isoenzymes Mbblh and Ssbco with comparable catalytic properties but different catalytic mechanisms.Combinational expression of isoenzymes increased the flux fromβ-carotene to vitamin A metabolism.To modulate the vitamin A components,retinol dehydrogenase 12 from Homo sapiens was introduced to achieve more than 90%retinol purity using shake flask fermentation.Overexpressing POS5Δ17 enhanced the reduced nicotinamide adenine dinucleotide phosphate pool,and the titer of vitamin A was elevated by almost 46%.Multi-copy integration of the key rate-limiting step gene Mbblh further improved the synthesis of vitamin A.Consequently,the titer of vitamin A in the strain harbouring the Ura3 marker was increased to 588 mg/L at the shake-flask level.Eventually,the highest reported titer of 5.21 g/L vitamin A in S.cerevisiae was achieved in a 1-L bioreactor.This study unlocked the potential of S.cerevisiae for synthesising vitamin A in a sustainable and economical way,laying the foundation for the commercial-scale production of bio-based vitamin A.展开更多
Lignin,a natural and renewable aromatic polymer,serves as a plentiful source of aromatic building blocks for biomanufacturing.However,despite its potential,the bioconversion efficiency of lignin remains limited due to...Lignin,a natural and renewable aromatic polymer,serves as a plentiful source of aromatic building blocks for biomanufacturing.However,despite its potential,the bioconversion efficiency of lignin remains limited due to its inherently complex structure and the constraints of traditional metabolic engineering approaches.Synthetic biology-guided metabolic regulation offers a solution by coordinating intracellular resource allocation in ligninolytic strains,endowing their adaptation for industrial applications and promoting the sustainability of lignin-based bioeconomy.This review provides a comprehensive overview of multiscale metabolic regulation,including critical enzymes involved in biotransformation,metabolic pathway networks,genome-phenotype,and learning prediction.It highlights the significant roles and potential benefits of emerging cutting-edge technologies in advancing lignin valorization.Overall,synthetic biology-guided metabolic regulation has demonstrated its power in balancing the metabolic fluxes of ligninolytic strains,ensuring the continued vitality in future development.展开更多
Corynebacterium glutamicum is a versatile industrial microorganism for producing various amino acids.However,there have been no reports of well-defined C.glutamicum strains capable of hyperproducing L-tryptophan.This ...Corynebacterium glutamicum is a versatile industrial microorganism for producing various amino acids.However,there have been no reports of well-defined C.glutamicum strains capable of hyperproducing L-tryptophan.This study presents a comprehensive metabolic engineering approach to establish robust C.glutamicum strains for Ltryptophan biosynthesis,including:(1)identification of potential targets by enzyme-constrained genome-scale modeling;(2)enhancement of the L-tryptophan biosynthetic pathway;(3)reconfiguration of central metabolic pathways;(4)identification of metabolic bottlenecks through comparative metabolome analysis;(5)engineering of the transport system,shikimate pathway,and precursor supply;and(6)repression of competing pathways and iterative optimization of key targets.The resulting C.glutamicum strain achieved a remarkable L-tryptophan titer of 50.5 g/L in 48h with a yield of 0.17 g/g glucose in fed-batch fermentation.This study highlights the efficacy of integrating computational modeling with systems metabolic engineering for significantly enhancing the production capabilities of industrial microorganisms.展开更多
Dear Editor,Astaxanthin,a potentantioxidant carotenoid,is widely used in aquaculture,nutraceuticals,and cosmetics(Schmidt et al.,2011).Over 95%of commercial astaxanthin is chemicallysynthesized:however,its low bioacti...Dear Editor,Astaxanthin,a potentantioxidant carotenoid,is widely used in aquaculture,nutraceuticals,and cosmetics(Schmidt et al.,2011).Over 95%of commercial astaxanthin is chemicallysynthesized:however,its low bioactivity and environmental concerns necessitate the sustainable biosynthetic alternatives(Jiang et al..2017).The biosynthesis of astaxanthin by Phaffia rhodozyma has been investigated in detail,primarily due to severaladvantages such as simple culture condition.展开更多
Dear Editor,Theanine(N-ethyl-γ-L-glutamine)is a characteristic amino acid primarily found in tea leaves(Camellia sinensis).It is derived from pyruvate through three enzymatic steps involving alanine aminotransferase(...Dear Editor,Theanine(N-ethyl-γ-L-glutamine)is a characteristic amino acid primarily found in tea leaves(Camellia sinensis).It is derived from pyruvate through three enzymatic steps involving alanine aminotransferase(AlaAT),alanine decarboxylase(AlaDC),and theanine synthetase(TS)(She et al.,2022;Luo and He,2025).Theanine has attracted widespread attention due to its tasteenhancing properties and potential health benefits,including relaxation and cognitive enhancement(Li et al.,2022),however,its limited natural production,coupled with increasing demand,has seriously restricted its broad application.展开更多
Riboflavin,an important vitamin utilized in pharmaceutical products and as a feed additive,is mainly produced by metabolically engineered bacterial fermentation.However,the reliance on antibiotics in the production pr...Riboflavin,an important vitamin utilized in pharmaceutical products and as a feed additive,is mainly produced by metabolically engineered bacterial fermentation.However,the reliance on antibiotics in the production process leads to increased costs and safety risks.To address these challenges,an antibiotic-free Escherichia coli riboflavin producer was constructed using metabolic engineering approaches coupled with a novel plasmid stabilization system.Initially,competitive pathways and feedback inhibition were attenuated to enhance the metabolic flux towards riboflavin.Key genes in the purine pathway were overexpressed to boost the availability of riboflavin precursors.Subsequently,a plasmid stabilization system based on toxin was screened and characterized,achieving a plasmid retention rate of 84.9%after 10 days of passaging.Finally,transcriptomic analysis at the genome-wide level revealed several rate-limiting genes,including pgl,gnd,and yigB,which were subsequently upregulated,leading to a 26%improvement in riboflavin production.With optimization of the culture medium,the final strain allowed the production of 11.5 g/L of riboflavin with a yield of 90.4 mg/g glucose in 5 L bioreactors without antibiotics.These strategies can be extended to other plasmid-based riboflavin derivative production systems.展开更多
Pneumocandin B_(0)(PB_(0))is a lipohexapeptide synthesized by Glarea lozoyensis and serves as the precursor for the widely used antifungal drug caspofungin acetate(Cancidas®).However,the low titer of PB_(0)result...Pneumocandin B_(0)(PB_(0))is a lipohexapeptide synthesized by Glarea lozoyensis and serves as the precursor for the widely used antifungal drug caspofungin acetate(Cancidas®).However,the low titer of PB_(0)results in fermentation and purification costs during caspofungin production,limiting its widespread clinical application.Here,we engineered an efficient PB_(0)-producing strain of G.lozoyensis by systems metabolic engineering strategies,including multi-omics analysis and multilevel metabolic engineering.We overexpressed four rate-limiting enzymes:thioesterase GLHYD,two cytochrome P450s GLP450s,and chorismate synthase GLCS;knocked out two competing pathways responsible for producing 6-methylsalicylic acid and pyranidine E;and overexpressed the global transcriptional activator GLHYP.As a result,the PB_(0)titer increased by 108.7%to 2.63 g/L at the shake-flask level through combinatorial strategies.Our study provides valuable insights into achieving high-level production of PB_(0)and offers general guidance for developing efficient fungal cell factories to produce polyketide synthase-non-ribosomal peptide synthetase hybrid metabolites.展开更多
Abscisic acid(ABA)is an important plant growth regulator with broad applications in agriculture,forestry,and other fields.Currently,the industrial production of ABA primarily relies on microbial fermentation using Bot...Abscisic acid(ABA)is an important plant growth regulator with broad applications in agriculture,forestry,and other fields.Currently,the industrial production of ABA primarily relies on microbial fermentation using Botrytis cinerea,but its genetic toolbox is limited.To address this,we first screened 10 strong constitutive promoters from the genome of B.cinerea through transcriptomic analysis.The expression levels of the promoters covered a range of 3-4 orders of magnitude according to the measured β-glucuronidase activity.Subsequently,four promoters of different strength were used to balance the cofactor supply in B.cinerea.Overexpression of NADH kinase using the medium-strength promoter Pef1a significantly enhanced ABA production,resulting in a 32.26%increase compared to the control.Finally,by combining promoter engineering with a push-pull strategy,we optimized the biosynthesis of ABA.The recombinant strain Pthi4:hmgr-Pef1a:a4,overexpressing HMGR under the Pthi4 promoter and Bcaba4 under the Pef1a promoter,achieved an ABA titer of 1.18 g/L,a 58.92%increase.To our best knowledge,this is the first constitutive promoter library suitable for B.cinerea,providing important tools for the industrial production of ABA.展开更多
基金supported by the Guangdong Provincial Key Research and Development Program-Modern Seed Industry,China(2022B0202060004)the Knowledge Innovation Program of Wuhan-Basic Research,China(2022020801010291)+1 种基金the Project of the Development for High-quality Seed Industry of Hubei Province,China(HBZY2023B003)the Innovation Program of the Chinese Academy of Agricultural Sciences(2023-2060299-089-031)。
文摘With the progress of urbanization,rural tourism has emerged as a popular leisure activity in China.The crop field with pattern art has been gaining popularity over the years,using strains with colorful leaves to create various impressive designs(Xu 2024).A lot of successful cases in rice fields have not only attracted tourists but also increased the income of farmers(Song et al.2020).Plants with colorful leaves also have gained significant popularity in ornamental agriculture.Therefore,developing plants with colorful leaves has gained significant popularity in ornamental agriculture.
基金supported by the Bio&Medical Technology Development Program of the National Research Foundation(NRF)funded by the Korean government(MSIT)(Grant No.RS-2024-00440478)to Sun-Hwa HAthe NRF by MSIT(Grant Nos.RS-2024-00347806 and RS-2024-00407469)to Sun-Hwa HAthe New Plant Breed Technology Program funded by the Rural Development Administration,Republic of Korea(Grant No.RS-2024-00322447)to Sun-Hwa HA.
文摘To improve the nutritional and functional value of rice,numerous biotechnological approaches have focused on metabolic engineering to address nutritional deficiencies and produce health-beneficial compounds that are either absent or naturally present in low amounts.A prominent example is‘Golden Rice’,which has been genetically modified to accumulateβ-carotene to combat vitamin A deficiency in regions with limited dietary intake.Scientists have been continuously biofortifying rice with various specialized metabolites,including terpenoids,flavonoids,non-flavonoid polyphenols,betalains,vitamins,and amino acids.This review explores the specific pathways and genetic modifications utilized by researchers to enhance the accumulation of targeted metabolites in rice.It comprehensively summarizes key strategies and research trends in rice metabolic engineering,demonstrating how rice can be transformed into a strategic crop for producing industrially valuable compounds beyond its traditional role as a staple food by leveraging its advantages as a versatile host system through its grains,leaves,and cells.Furthermore,we highlight the potential of intergrating metabolic engineering with synthetic biology and big data-driven computational modeling,particularly through artificial intelligence and machine learning,as promising future research directions.
文摘Tyrosol is a natural phenolic compound with antioxidant,anti-inflammatory and other biological activities,serving as an important precursor of high-value products such as hydroxytyrosol and salidroside.Therefore,the green and efficient biosynthesis of tyrosol and its derivatives has become a research hotspot in recent years.Building cell factories by metabolic engineering of microorganisms is a potential industrial production way,which has low costs and environmental friendliness.This paper introduces the biosynthesis pathway of tyrosol and presents the key regulated nodes in the de novo synthesis of tyrosol in Escherichia coli and Saccharomyces cerevisiae.In addition,this paper reviews the recent advances in metabolic engineering for the production of hydroxytyrosol and salidroside.This review can provide a reference for engineering the strains for the high-yield production of tyrosol and its derivatives.
基金by grants from the National Research Foundation,Ministry of Science and ICT,Republic of Korea(NRF-2021R1A2B5B03002123,NRF-2018R1A5A2024425,NRF-2021K2A9A2A06044515,2022M3E5F1017919)Ministry of Education,Republic of Korea(NRF-2021R1A6A3A01086428)Korean Health Technology R&D Project(No.HI19C0664),Ministry of Health&Welfare,Republic of Korea.
文摘Metabolic engineering of the tumor microenvironment has emerged as a new strategy.Lactate dehydrogenase A(LDHA)is a prominent target for metabolic engineering.Here,we designed a cationic lipid nanoparticle formulation for LDHA gene editing.The plasmid DNA delivery efficiency of our lipid nanoparticle formulations was screened by testing the fluorescence of lipid nanoparticles complexed to plasmid DNA encoding green fluorescence protein(GFP).The delivery efficiency was affected by the ratios of three components:a cationic lipid,cholesterol or its derivative,and a fusogenic lipid.The lipid nanoparticle designated formulation F3 was complexed to plasmid DNA co-encoding CRISPR-associated protein 9 and LDHA-specific sgRNA,yielding the lipoplex,pCas9-sgLDHA/F3.The lipoplex including GFP-encoding plasmid DNA provided gene editing in HeLa-GFP cells.Treatment of B16F10 tumor cells with pCas9-sgLDHA/F3 yielded editing of the LDHA gene and increased the pH of the culture medium.pCas9-sgLDHA/F3 treatment activated the interferon-gamma and granzyme production of T cells in culture.In vivo,combining pCas9-sgLDHA/F3 with immune checkpoint-inhibiting anti-PD-L1 antibody provided a synergistic antitumor effect and prolonged the survival of tumor model mice.This study suggests that combining metabolic engineering of the tumor microenvironment with immune checkpoint inhibition could be a valuable antitumor strategy.
基金The research was financially supported by the National Natural Science Foundation of China(NSFC-21776209,NSFC-21621004,NSFC-21776208)Natural Science Foundation of Tianjin(No.19JCYBJC21100).
文摘In recent years,metabolic engineering has made great progress in both academic research and industrial applications.However,we have not found any articles that specifically analyze the current state of metabolic engineering in China in comparison with other countries.Here,we review the current development and future trends of global metabolic engineering,conduct an in-depth benchmarking analysis of the development situation of China’s metabolic engineering,and identify current problems as well as future trends.We searched publications in the Scopus database from 2015 to September 2020 in the field of metabolic engineering,and analyzed the output in general,including publication trends,research distribution,popular journals,hot topics and vital institutions,but also analyzed the share of citations,field-weighted citation impact,and production in collaboration with strategic countries in science and technology.This study aims to serve as a reference for later studies,offering a comprehensive view of China’s contribution to metabolic engineering,and as a tool for the elaboration of national public policy in science and technology.
文摘Through several waves of technological research and un‐matched innovation strategies,bio‐catalysis has been widely used at the industrial level.Because of the value of enzymes,methods for producing value‐added compounds and industrially‐relevant fine chemicals through biological methods have been developed.A broad spectrum of numerous biochemical pathways is catalyzed by enzymes,including enzymes that have not been identified.However,low catalytic efficacy,low stability,inhibition by non‐cognate substrates,and intolerance to the harsh reaction conditions required for some chemical processes are considered as major limitations in applied bio‐catalysis.Thus,the development of green catalysts with multi‐catalytic features along with higher efficacy and induced stability are important for bio‐catalysis.Implementation of computational science with metabolic engineering,synthetic biology,and machine learning routes offers novel alternatives for engineering novel catalysts.Here,we describe the role of synthetic biology and metabolic engineering in catalysis.Machine learning algorithms for catalysis and the choice of an algorithm for predicting protein‐ligand interactions are discussed.The importance of molecular docking in predicting binding and catalytic functions is reviewed.Finally,we describe future challenges and perspectives.
基金supported by the Key Research and Development Program for Science and Technology of Jinzhong(No.Y191021)the Basic Research Project of Shanxi University of Traditional Chinese Medicine(No.2020PY-JC(Y)-05)the Research Project Supported by Shanxi Scholarship Council of China(No.2020-132)。
文摘Mushrooms are abundant in bioactive natural compounds.Due to strict growth conditions and long fermentation-time,microbe as a production host is an alternative and sustainable approach for the production of natural compounds.This review focuses on the biosynthetic pathways of mushroom originated natural compounds and microbes as the production host for the production of the above natural compounds.
基金the National Natural Science Foundation of China(32270586,31825019,and 31801322)the Department of Science and Technology of Jiangsu Province(BM2022008-02 and BE2022336).
文摘In animals,serotonin is a neurotransmitter and mood regulator.In plants,serotonin functions in energy acquisition,tissue maintenance,delay of senescence,and response to biotic and abiotic stresses.In this study,we examined the effect of serotonin enrichment of rice endosperm on plant growth,endosperm development,and grain quality.To do so,TDCs and T5H were selected as targets for serotonin fortification.Overexpression of TDC1 or TDC3 increased serotonin accumulation relative to overexpression of T5H in rice grain.Transgenic lines of target genes driven by the Gt1 promoter showed better field performance than those driven by the Ubi promoter.Overexpression of T5H showed little effect on plant growth or grain physicochemical quality.In neuronal cell culture assays,serotonin induced neuroprotective action against apoptosis.Breeding of rice cultivars with high serotonin content may be beneficial for health and nutrition.
基金partially supported by Department of Science and Technology,Science and Engineering Research Board under Teachers Associateship for Research Excellence(TARE)Scheme(File Number TAR/2023/000036).
文摘Depleting global petroleum reserves and skyrocketing prices coupled with succinct supply have been a grave concern,which needs alternative sources to conventional fuels.Oleaginous microalgae have been explored for enhanced lipid production,leading towards biodiesel production.These microalgae have short life cycles,require less labor,and space,and are easy to scale up.Triacylglycerol,the primary source of lipids needed to produce biodiesel,is accumulated by most microalgae.The article focuses on different types of oleaginous microalgae,which can be used as a feedstock to produce biodiesel.Lipid biosynthesis in microalgae occurs through fatty acid synthesis and TAG synthesis approaches.In-depth discussions are held regarding other efficient methods for enhancing fatty acid and TAG synthesis,regulating TAG biosynthesis bypass methods,blocking competing pathways,multigene approach,and genome editing.The most potential targets for gene transformation are hypothesized to be a malic enzyme and diacylglycerol acyltransferase while lowering phosphoenolpyruvate carboxylase activity is reported to be advantageous for lipid synthesis.
文摘Mesaconic acid has a special chemical structure and can undergo a series of reactions such as polymerization and addition. It is an important chemical intermediate and widely used in material, chemical and other industries. The chemical synthesis of mesaconic acid requires nitric acid, which is dangerous and harmful to the environment. The production of mesaconic acid by microbial fermentation has the characteristics of low raw material price, high efficiency and strong specificity, and thus a strong industrial application prospect. Mesaconic acid is an intermediate product of glutamic acid degradation pathway of microorganisms such as Clostridium tetani. However, at present, few reports have been conducted on the production of mesaconic acid by metabolic engineering microorganisms. In this study, glutamate mutase(GLM) and 3-methylaspartate ammonialyase(MAL) from C. tetani were recombined and expressed in Escherichia coli, and the obtained strain, BL21(DE3)/pETDuet-1-MAL-mutS-mutE, achieved the yield of mesaconic acid of 1.06 g/L. Compared with the wild type, the yields of mesaconic acid from mutants G133A and G133S increased by 21% and 16%, respectively. After 24 h of flask fermentation, the yields of mesaconic acid reached 1.28 and 1.23 g/L, respectively. This study can provide reference for microbial synthesis of mesaconic acid.
基金Funding for this research was graciously provided by several key sources:the National Key Research and Development Program of China under grant number 2019YFA0905000the Fundamental Research Funds for the Central Universities,grant number 222201714026the National Natural Science Foundation of China with grant numbers 21871085 and 31971380.
文摘Biosynthesis of paclitaxel(Taxol™)is a hot topic with extensive and durable interests for decades.However,it is severely hindered due to the very low titers of intermediates.In this study,Escherichia coli was employed to de novo synthesize a key intermediate of paclitaxel,taxadien-5α-yl-acetate(T5OAc).Plasmid-based pathway reconstruction and optimization were conducted for T5OAc production.The endogenous methylerythritol phosphate pathway was enhanced to increase the precursor supply.Three taxadien-5α-ol O-acetyltransferases were tested to obtain the best enzyme for the acetylation step.Metabolic burden was relieved to restore cell growth and promote production through optimizing the plasmid production system.In order to achieve metabolic balance,the biosynthesis pathway was regulated precisely by multivariate-modular metabolic engineering.Finally,in a 5-L bioreactor,the T5OAc titer was enhanced to reach 10.9 mg/L.This represents an approximately 272-fold increase in production compared to the original strain,marking the highest yield of T5OAc ever documented in E.coli,which is believed to be helpful for promoting the progress of paclitaxel biosynthesis.
文摘Objective:The biosynthesis of pilose antler polysaccharides(PAPS)with anti-aging activity relies on the precise regulation of key enzymes;however,the identification of these enzymes is limited by traditional low-throughput techniques.This study constructed an integrated system combining metabolic engineering and high-throughput screening of fluorescent substrates,aiming to overcome the technical bottlenecks in the identification of key enzymes for PAPS synthesis and the optimization of their catalytic efficiency.Methods:Eighteen candidate genes related to carbohydrate metabolism were screened via transcriptome sequencing of deer antler stem cells.After prokaryotic expression and preliminary screening by high-performance liquid chromatography(HPLC),a self-designed fluorescent substrate(FAM-Glc-β1-3-PNPG)combined with a 96-well plate platform was used to achieve high-throughput optimization and screening of catalytic efficiency.Enzyme function was verified through kinetic analysis and in vitro antioxidant experiments.Results:The screened high-efficiency glycosyltransferase UGT-Wnt3a showed a 2.1-fold increase in catalytic efficiency compared with the wild type,with a maximum reaction rate(Vmax)of12.3μmol·min^(-1)·mg^(-1)and a Michaelis constant(Km)of 0.87 mM.The catalytic product of UGT-Wnt3a increased the superoxide dismutase(SOD)activity by 42%(P<0.01)and decreased the malondialdehyde(MDA)content by 28%(P<0.05)in the oxidative damage system.The detection efficiency of the new platform was 9 times higher than that of HPLC,and the limit of detection was reduced by 50 times.Conclusion:The screening system established in this study provides an innovative technical solution for the directed synthesis of PAPS and the development of anti-aging components,promoting the transformation of active components in traditional Chinese medicine from natural extraction to bioengineering-based production.
基金supported by the National Key Research and Development Program(2023YFF1103701)the Key-Area Research and Development Program of Guangdong Province(2020B0303070002)+1 种基金the National Key Research and Development Program of China(2018YFA0900702)the National Natural Science Foundation of China(22178261).
文摘Vitamin A is a micronutrient critical for versatile biological functions and has been widely used in the food,cosmetics,pharmaceutical,and nutraceutical industries.Synthetic biology and metabolic engineering enable microbes,especially the model organism Saccharomyces cerevisiae(generally recognised as safe)to possess great potential for the production of vitamin A.Herein,we first generated a vitamin A-producing strain by miningβ-carotene 15,15′-mono(di)oxygenase from different sources and identified two isoenzymes Mbblh and Ssbco with comparable catalytic properties but different catalytic mechanisms.Combinational expression of isoenzymes increased the flux fromβ-carotene to vitamin A metabolism.To modulate the vitamin A components,retinol dehydrogenase 12 from Homo sapiens was introduced to achieve more than 90%retinol purity using shake flask fermentation.Overexpressing POS5Δ17 enhanced the reduced nicotinamide adenine dinucleotide phosphate pool,and the titer of vitamin A was elevated by almost 46%.Multi-copy integration of the key rate-limiting step gene Mbblh further improved the synthesis of vitamin A.Consequently,the titer of vitamin A in the strain harbouring the Ura3 marker was increased to 588 mg/L at the shake-flask level.Eventually,the highest reported titer of 5.21 g/L vitamin A in S.cerevisiae was achieved in a 1-L bioreactor.This study unlocked the potential of S.cerevisiae for synthesising vitamin A in a sustainable and economical way,laying the foundation for the commercial-scale production of bio-based vitamin A.
基金supported by the National Key Research and Development Program of China(2023YFC3403500).
文摘Lignin,a natural and renewable aromatic polymer,serves as a plentiful source of aromatic building blocks for biomanufacturing.However,despite its potential,the bioconversion efficiency of lignin remains limited due to its inherently complex structure and the constraints of traditional metabolic engineering approaches.Synthetic biology-guided metabolic regulation offers a solution by coordinating intracellular resource allocation in ligninolytic strains,endowing their adaptation for industrial applications and promoting the sustainability of lignin-based bioeconomy.This review provides a comprehensive overview of multiscale metabolic regulation,including critical enzymes involved in biotransformation,metabolic pathway networks,genome-phenotype,and learning prediction.It highlights the significant roles and potential benefits of emerging cutting-edge technologies in advancing lignin valorization.Overall,synthetic biology-guided metabolic regulation has demonstrated its power in balancing the metabolic fluxes of ligninolytic strains,ensuring the continued vitality in future development.
基金supported by the National Key R&D Program of China(No.2021YFC2100900)the National Natural Science Foundation of China(Grant Nos.21938004,22078172)Tsinghua University Initiative Scientific Research Program(No.20223080016).
文摘Corynebacterium glutamicum is a versatile industrial microorganism for producing various amino acids.However,there have been no reports of well-defined C.glutamicum strains capable of hyperproducing L-tryptophan.This study presents a comprehensive metabolic engineering approach to establish robust C.glutamicum strains for Ltryptophan biosynthesis,including:(1)identification of potential targets by enzyme-constrained genome-scale modeling;(2)enhancement of the L-tryptophan biosynthetic pathway;(3)reconfiguration of central metabolic pathways;(4)identification of metabolic bottlenecks through comparative metabolome analysis;(5)engineering of the transport system,shikimate pathway,and precursor supply;and(6)repression of competing pathways and iterative optimization of key targets.The resulting C.glutamicum strain achieved a remarkable L-tryptophan titer of 50.5 g/L in 48h with a yield of 0.17 g/g glucose in fed-batch fermentation.This study highlights the efficacy of integrating computational modeling with systems metabolic engineering for significantly enhancing the production capabilities of industrial microorganisms.
基金supported by the National key Research and Development Program of China(2022YFC2105002).
文摘Dear Editor,Astaxanthin,a potentantioxidant carotenoid,is widely used in aquaculture,nutraceuticals,and cosmetics(Schmidt et al.,2011).Over 95%of commercial astaxanthin is chemicallysynthesized:however,its low bioactivity and environmental concerns necessitate the sustainable biosynthetic alternatives(Jiang et al..2017).The biosynthesis of astaxanthin by Phaffia rhodozyma has been investigated in detail,primarily due to severaladvantages such as simple culture condition.
基金supported by the Key Areas Research Projects of Guangdong Province(2022B0202060002)the Incubation Program for the Science and Technology Development of Chinese Medicine Guangdong Laboratory(HQL2024PZ028)+4 种基金the Guangdong Basic and Applied Basic Research Foundation(2024A1515012120)the Guangzhou Basic and Applied Basic Research Foundation(2023A04J00359)the Special Foundation for Introduction of Scientific Talents of GDAAS(R2021YJ-YB3015,R2022YJ-YB1003,R2022PY-QY001,and R2021YJ-XD001)the Elite Rice Plan of GDRRI(2022YG01,2022YG02,and 2025YG02)the Guangdong Key Laboratory of Rice Science and Technology(2023B1212060042).
文摘Dear Editor,Theanine(N-ethyl-γ-L-glutamine)is a characteristic amino acid primarily found in tea leaves(Camellia sinensis).It is derived from pyruvate through three enzymatic steps involving alanine aminotransferase(AlaAT),alanine decarboxylase(AlaDC),and theanine synthetase(TS)(She et al.,2022;Luo and He,2025).Theanine has attracted widespread attention due to its tasteenhancing properties and potential health benefits,including relaxation and cognitive enhancement(Li et al.,2022),however,its limited natural production,coupled with increasing demand,has seriously restricted its broad application.
基金supported by the National Key Research and Devel-opment Program of China(No.2021YFC2100900,2022YFA0912200)the National Natural Science Foundation of China(No.32071470,32300063)+1 种基金the Natural Science Foundation of Jiangsu Province(No.BK20221080,No.BK20210464)the Research Program of State Key Laboratory of Food Science and Resources,Jiangnan University(No.SKLF-ZZB-202408).
文摘Riboflavin,an important vitamin utilized in pharmaceutical products and as a feed additive,is mainly produced by metabolically engineered bacterial fermentation.However,the reliance on antibiotics in the production process leads to increased costs and safety risks.To address these challenges,an antibiotic-free Escherichia coli riboflavin producer was constructed using metabolic engineering approaches coupled with a novel plasmid stabilization system.Initially,competitive pathways and feedback inhibition were attenuated to enhance the metabolic flux towards riboflavin.Key genes in the purine pathway were overexpressed to boost the availability of riboflavin precursors.Subsequently,a plasmid stabilization system based on toxin was screened and characterized,achieving a plasmid retention rate of 84.9%after 10 days of passaging.Finally,transcriptomic analysis at the genome-wide level revealed several rate-limiting genes,including pgl,gnd,and yigB,which were subsequently upregulated,leading to a 26%improvement in riboflavin production.With optimization of the culture medium,the final strain allowed the production of 11.5 g/L of riboflavin with a yield of 90.4 mg/g glucose in 5 L bioreactors without antibiotics.These strategies can be extended to other plasmid-based riboflavin derivative production systems.
基金funded by the National Key R&D Program of China(2021YFC2102600)the National Natural Science Foundation of China(32370060 and 32070063)the Shenzhen Science and Technology Program(JCYJ20241202124932044).
文摘Pneumocandin B_(0)(PB_(0))is a lipohexapeptide synthesized by Glarea lozoyensis and serves as the precursor for the widely used antifungal drug caspofungin acetate(Cancidas®).However,the low titer of PB_(0)results in fermentation and purification costs during caspofungin production,limiting its widespread clinical application.Here,we engineered an efficient PB_(0)-producing strain of G.lozoyensis by systems metabolic engineering strategies,including multi-omics analysis and multilevel metabolic engineering.We overexpressed four rate-limiting enzymes:thioesterase GLHYD,two cytochrome P450s GLP450s,and chorismate synthase GLCS;knocked out two competing pathways responsible for producing 6-methylsalicylic acid and pyranidine E;and overexpressed the global transcriptional activator GLHYP.As a result,the PB_(0)titer increased by 108.7%to 2.63 g/L at the shake-flask level through combinatorial strategies.Our study provides valuable insights into achieving high-level production of PB_(0)and offers general guidance for developing efficient fungal cell factories to produce polyketide synthase-non-ribosomal peptide synthetase hybrid metabolites.
基金supported by National key research and development program of China(2021YFC2104300)Project of Leading Innovative Talents in Short-term Jiangxi Province(jxsq2023102172)+1 种基金Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project(TSBICIP-PTJS-003-04)Postgraduate Research&Practice Innovation Program of Jiangsu Province KYCX23_1750.
文摘Abscisic acid(ABA)is an important plant growth regulator with broad applications in agriculture,forestry,and other fields.Currently,the industrial production of ABA primarily relies on microbial fermentation using Botrytis cinerea,but its genetic toolbox is limited.To address this,we first screened 10 strong constitutive promoters from the genome of B.cinerea through transcriptomic analysis.The expression levels of the promoters covered a range of 3-4 orders of magnitude according to the measured β-glucuronidase activity.Subsequently,four promoters of different strength were used to balance the cofactor supply in B.cinerea.Overexpression of NADH kinase using the medium-strength promoter Pef1a significantly enhanced ABA production,resulting in a 32.26%increase compared to the control.Finally,by combining promoter engineering with a push-pull strategy,we optimized the biosynthesis of ABA.The recombinant strain Pthi4:hmgr-Pef1a:a4,overexpressing HMGR under the Pthi4 promoter and Bcaba4 under the Pef1a promoter,achieved an ABA titer of 1.18 g/L,a 58.92%increase.To our best knowledge,this is the first constitutive promoter library suitable for B.cinerea,providing important tools for the industrial production of ABA.
