Synthetic and Systems Biotechnology最新文献

{"title":"Optimizing longifolene production in Yarrowia lipolytica via metabolic and protein engineering","authors":"Yi-Tong Yao ,&nbsp;Xiao Zhang ,&nbsp;Chen-Yu Wang,&nbsp;Yu-He Zhang,&nbsp;Da-Wei Li,&nbsp;Wei-Dong Yang,&nbsp;Hong-Ye Li,&nbsp;Li-Gong Zou","doi":"10.1016/j.synbio.2025.01.004","DOIUrl":"10.1016/j.synbio.2025.01.004","url":null,"abstract":"<div><div>Longifolene (C₁₅H₂₄) is a tricyclic sesquiterpene widely utilized in the cosmetics and fragrances due to its versatile applications. Traditional extraction methods from plants suffer from low titer and lengthy production cycles, while chemical synthesis is hampered by the compound's complex structure, leading to high costs and insufficient market supply. This study aimed to develop a microbial cell factory for enhanced longifolene production. The strategy involved integrating longifolene synthase from <em>Pinus sylvestris</em> (<em>PsTPS</em>) into <em>Yarrowia lipolytica</em> and employing multiple metabolic engineering approaches. Initially, key genes in the mevalonate (MVA) pathway were overexpressed to enhance longifolene precursor availability for longifolene biosynthesis. Subsequently, protein engineering techniques were applied to optimize <em>PsTPS</em> (t<em>PsTPS</em>) for improved catalytic efficiency. Furthermore, co-expression of molecular chaperones was implemented to enhance the synthesis and secretion of <em>PsTPS</em>. The introduction of the isopentenol utilization pathway (IUP) further augmented the supply of C5 substrate. By optimizing the culture conditions, including a reduction in culture temperature, the efflux of longifolene was increased, and the dissolved oxygen levels were enhanced to promote the growth of the strain. These collective efforts resulted culminated in the engineered strain Z03 achieving a noteworthy production level of 34.67 mg/L of longifolene in shake flasks. This study not only demonstrates the feasibility of enhancing sesquiterpene production in <em>Y. lipolytica</em> but also highlights the potential of microbial platforms in meeting industrial demands for complex natural products.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 2","pages":"Pages 433-441"},"PeriodicalIF":4.4,"publicationDate":"2025-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143161057","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhancing rufomycin production by CRISPR/Cas9-based genome editing and promoter engineering in Streptomyces sp. MJM3502","authors":"Chun Su ,&nbsp;Nguyen-Quang Tuan ,&nbsp;Wen-Hua Li ,&nbsp;Jin-Hua Cheng ,&nbsp;Ying-Yu Jin ,&nbsp;Soon-Kwang Hong ,&nbsp;Hyun Lee ,&nbsp;Mallique Qader ,&nbsp;Larry Klein ,&nbsp;Gauri Shetye ,&nbsp;Guido F. Pauli ,&nbsp;Scott G. Flanzblau ,&nbsp;Sang-Hyun Cho ,&nbsp;Xin-Qing Zhao ,&nbsp;Joo-Won Suh","doi":"10.1016/j.synbio.2025.01.002","DOIUrl":"10.1016/j.synbio.2025.01.002","url":null,"abstract":"<div><div><em>Streptomyces</em> sp. MJM3502 is a promising producer of rufomycins, which are a class of potent anti-tuberculosis lead compounds. Although the structure, activity, and mechanism of the main rufomycin 4/6 and its analogs have been extensively studied, a significant gap remains in our understanding of the genome sequence and biosynthetic pathway of <em>Streptomyces</em> sp. MJM3502, and its metabolic engineering has not yet been reported. This study established the genetic manipulation platform for the strain. Using CRISPR/Cas9-based technology to in-frame insert the strong <em>kasO∗p</em> promoter upstream of the <em>rufB</em> and <em>rufS</em> genes of the rufomycin BGC, we increased rufomycin 4/6 production by 4.1-fold and 2.8-fold, respectively. Furthermore, designing recombinant strains by inserting the <em>kasO∗p</em> promoter upstream of the biosynthetic genes encoding cytochrome P450 enzymes led to new rufomycin derivatives. These findings provide the basis for enhancing the production of valuable natural compounds in <em>Streptomyces</em> and offer insights into the generation of novel active natural products via synthetic biology and metabolic engineering.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 2","pages":"Pages 421-432"},"PeriodicalIF":4.4,"publicationDate":"2025-01-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143160997","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Proton motive force generated by microbial rhodopsin promotes extracellular electron transfer","authors":"Wenqi Ding ,&nbsp;Tong Lin ,&nbsp;Yun Yang ,&nbsp;Wen-Wei Li ,&nbsp;Shaoan Cheng ,&nbsp;Hao Song","doi":"10.1016/j.synbio.2025.01.001","DOIUrl":"10.1016/j.synbio.2025.01.001","url":null,"abstract":"<div><div>The primary limitation to the practicability of electroactive microorganisms in bioelectrochemical systems lies in their low extracellular electron transfer (EET) efficiency. The proton motive force (PMF) represents the electrochemical gradient of protons generated by electron transport and proton pumping across the cytoplasmic membrane, serving as a crucial energy transfer pathway in bacterial membranes. Nevertheless, the impact of PMF on the EET efficiency remains ambiguous, while the microbial rhodopsin offers a simple and efficient avenue for non-photosynthetic cells to harness PMF. Here, we studied the function of three microbial rhodopsins (Arch, Mac, and cR-1) in facilitating EET via their heterologous expression in <em>S. oneidensis</em>, a model electroactive microorganism. Among these, the recombinant strain expressing rhodopsin cR-1 exhibited the highest output power density of 0.87 W/m², 3.49-fold increase over the wild-type <em>S. oneidensis</em> MR-1. Our further transcriptomics analyses of the energy and materials metabolism of strain cR-1 showed that the underlying mechanism of enhanced EET efficiency was resulted from heterologous expression of the light-driven proton pump. The results suggested that strain cR-1 effectively expels protons to generate additional PMF and provide extra ATP supply to the cells, which facilitated lactate uptake and utilization, thus enhancing electrons generation in cells. This augmented intracellular electron pool capacity ultimately resulted in enhancement of EET rate and power generation efficiency of the recombinant <em>S. oneidensis</em>.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 2","pages":"Pages 410-420"},"PeriodicalIF":4.4,"publicationDate":"2025-01-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11786069/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143081010","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ergothioneine biosynthesis: The present state and future prospect","authors":"Li Liang,&nbsp;Xu Shan-Shan,&nbsp;Jiang Yan-Jun","doi":"10.1016/j.synbio.2024.10.008","DOIUrl":"10.1016/j.synbio.2024.10.008","url":null,"abstract":"<div><div>Ergothioneine (ERG), a rare natural thio-histidine derivative with potent antioxidant properties and diverse biological functions, is widely utilized in food processing, cosmetics, pharmaceuticals, and nutritional supplements. Current bioproduction methods for ERG primarily depend on fermenting edible mushrooms. However, with the advancement in synthetic biology, an increasing number of genetically engineered microbial hosts have been developed for ERG production, including <em>Escherichia coli</em>, <em>Saccharomyces cerevisiae</em>, and <em>Corynebacterium glutamicum</em>. Given the involvement of multiple precursor substances in ERG synthesis, it is crucial to employ diverse strategies to regulate the metabolic flux of ERG synthesis. This review comprehensively evaluates the physiological effects and safety considerations associated with ERG, along with the recent advancements in catalytic metabolic pathway for ERG production using synthetic biology tools. Finally, the review discusses the challenges in achieving efficient ERG production and the strategies to address these challenges using synthetic biology tools. This review provides a literature analysis and strategies guidance for the further application of novel synthetic biology tools and strategies to improve ERG yield.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 1","pages":"Pages 314-325"},"PeriodicalIF":4.4,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11664081/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142883028","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Regulatory significance of terminator: A systematic approach for dissecting terminator-mediated enhancement of upstream mRNA stability","authors":"Jia-Wei Ren ,&nbsp;Jin-Peng Zhang ,&nbsp;Zi-Lun Mei ,&nbsp;Jia-Yi Shao ,&nbsp;Guo-Qiang Xu ,&nbsp;Hui Li ,&nbsp;Jin-Song Gong ,&nbsp;Xiao-Mei Zhang ,&nbsp;Jin-Song Shi ,&nbsp;Xiao-juan Zhang ,&nbsp;Zheng-hong Xu","doi":"10.1016/j.synbio.2024.11.006","DOIUrl":"10.1016/j.synbio.2024.11.006","url":null,"abstract":"<div><div>The primary function of terminators is to terminate transcription in gene expression. Although some studies have suggested that terminators also contribute positively to upstream gene expression, the extent and underlying mechanism of this effect remain largely unexplored. Here, the correlation between terminating strength and upstream mRNA stability was investigated by constructing a terminator mutation library through randomizing 5 nucleotides, assisted by FlowSeq technology, terminator variants were categorized based on the downstream fluorescence intensity, followed by high-throughput sequencing. To examine the impact of terminators on mRNA stability, the abundance of downstream gene transcripts for each terminator variant was quantified through cDNA sequencing. The results revealed that the transcript abundance controlled by strong terminators was, on average 2.2 times greater than those controlled by weak terminators on average. Moreover, several distinct features could be ascribed to high relative abundance of upstream gene transcript, including a high GC content at the base region of hairpin, and a high AT content in downstream of the U-tract. Additionally, these terminators showed a free energy between −28 and −22 kcal/mol, and a stem length of 14 nt. Finally, these features ascribed the upstream beneficial terminator were validated across various expression systems. By incorporating the optimal terminator downstream of RSF, GSH and HIS in three different strains, the fermentation productions-NMN SAM and VD13 exhibited a remarkable enhancement of 30 %–70 %. The findings presented here uncovered the terminator characteristics contributed to the upstream mRNA stability, providing guiding principles for gene circuit design.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 1","pages":"Pages 326-335"},"PeriodicalIF":4.4,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11696848/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142932451","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"High-level production of free fatty acids from lignocellulose hydrolysate by co-utilizing glucose and xylose in yeast","authors":"Xin Ni ,&nbsp;Jingjing Li ,&nbsp;Wei Yu ,&nbsp;Fan Bai ,&nbsp;Zongbao K. Zhao ,&nbsp;Jiaoqi Gao ,&nbsp;Fan Yang ,&nbsp;Yongjin J. Zhou","doi":"10.1016/j.synbio.2024.12.009","DOIUrl":"10.1016/j.synbio.2024.12.009","url":null,"abstract":"<div><div>Lignocellulose bio-refinery via microbial cell factories for chemical production represents a renewable and sustainable route in response to resource starvation and environmental concerns. However, the challenges associated with the co-utilization of xylose and glucose often hinders the efficiency of lignocellulose bioconversion. Here, we engineered yeast <em>Ogataea polymorpha</em> to effectively produce free fatty acids from lignocellulose. The non-oxidative branch of the pentose phosphate pathway, and the adaptive expression levels of xylose metabolic pathway genes <em>XYL1</em>, <em>XYL2</em> and <em>XYL3</em>, were systematically optimized. In addition, the introduction of xylose transporter and global regulation of transcription factors achieved synchronous co-utilization of glucose and xylose. The engineered strain produced 11.2 g/L FFAs from lignocellulose hydrolysates, with a yield of up to 0.054 g/g. This study demonstrated that metabolic rewiring of xylose metabolism could support the efficient co-utilization of glucose and xylose from lignocellulosic resources, which may provide theoretical reference for lignocellulose biorefinery.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 2","pages":"Pages 401-409"},"PeriodicalIF":4.4,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11758827/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143047957","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Coupling genome-wide continuous perturbation with biosensor screening reveals the potential targets in yeast isopentanol synthesis network","authors":"Qi Xiao ,&nbsp;Jingjing Shi ,&nbsp;Lixian Wang ,&nbsp;Guoping Zhao ,&nbsp;Yanfei Zhang","doi":"10.1016/j.synbio.2024.12.010","DOIUrl":"10.1016/j.synbio.2024.12.010","url":null,"abstract":"<div><div>The increasing consumption of fossil fuels is contributing to global resource depletion and environmental pollution. Branched-chain higher alcohols, such as isopentanol and isobutanol, have attracted significant attention as next-generation biofuels. Biofuel production through microbial fermentation offers a green, sustainable, and renewable alternative to chemical synthesis. While enhanced production of isopentanol has been achieved in a variety of chassis, the fermentation yield has not yet reached levels suitable for industrial-scale production. In this study, we employed a continuous perturbation tool to construct a genome-scale perturbation library, combined with an isopentanol biosensor to screen for high-yielding mutants. We identified five high-yielding mutants, each exhibiting an increased glucose conversion rate and isopentanol titer. The F2 strain, in particular, achieved an isopentanol titer of 1.57 ± 0.014 g/L and a yield of 14.04 ± 0.251 mg/g glucose (10% glucose), surpassing the highest values reported to date in engineered <em>Saccharomyces cerevisiae</em>. Systematic transcriptome analysis of the isopentanol synthesis, glycolysis, glycerol metabolism, and ethanol synthesis pathways revealed that <em>MPC</em>, <em>OAC1</em>, <em>BAT2</em>, <em>GUT2</em>, <em>PDC6</em>, and <em>ALD4</em> are linked to efficient isopentanol production. Further analysis of differentially expressed genes (DEGs) identified 17 and 12 co-expressed DEGs (co-DEGs) in all mutants and the two second-round mutants, respectively. In addition, we validated the knockout or overexpression of key co-DEGs. Our results confirmed the critical roles of <em>HOM3</em> and <em>DIP5</em> in isopentanol production, along with genes associated with the aerobic respiratory chain (<em>SDH3</em>, <em>CYT1</em>, <em>COX7</em>, <em>ROX1</em>, and <em>ATG41</em>) and cofactor balance (<em>BNA2</em> and <em>NDE1</em>). Additionally, functional analysis of the co-DEGs revealed that <em>MAL33</em> is associated with the synthesis of branched-chain higher alcohols, expanding the intracellular metabolic network and offering new possibilities for green, cost-effective biofuel production.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 2","pages":"Pages 452-462"},"PeriodicalIF":4.4,"publicationDate":"2024-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143161058","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Design and structure of overlapping regions in PCA via deep learning","authors":"Yan Zheng ,&nbsp;Xi-Chen Cui ,&nbsp;Fei Guo ,&nbsp;Ming-Liang Dou ,&nbsp;Ze-Xiong Xie ,&nbsp;Ying-Jin Yuan","doi":"10.1016/j.synbio.2024.12.007","DOIUrl":"10.1016/j.synbio.2024.12.007","url":null,"abstract":"<div><div>Polymerase cycling assembly (PCA) stands out as the predominant method in the synthesis of kilobase-length DNA fragments. The design of overlapping regions is the core factor affecting the success rate of synthesis. However, there still exists DNA sequences that are challenging to design and construct in the genome synthesis. Here we proposed a deep learning model based on extensive synthesis data to discern latent sequence representations in overlapping regions with an AUPR of 0.805. Utilizing the model, we developed the SmartCut algorithm aimed at designing oligonucleotides and enhancing the success rate of PCA experiments. This algorithm was successfully applied to sequences with diverse synthesis constraints, 80.4 % of which were synthesized in a single round. We further discovered structure differences represented by major groove width, stagger, slide, and centroid distance between overlapping and non-overlapping regions, which elucidated the model's reasonableness through the lens of physical chemistry. This comprehensive approach facilitates streamlined and efficient investigations into the genome synthesis.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 2","pages":"Pages 442-451"},"PeriodicalIF":4.4,"publicationDate":"2024-12-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143161542","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Genetically-modified activation strategy facilitates the discovery of sesquiterpene-derived metabolites from Penicillium brasilianum","authors":"Wenni He ,&nbsp;Xiaoting Rong ,&nbsp;Hui Lv ,&nbsp;Lihua Zhang ,&nbsp;Jinglin Bai ,&nbsp;Lu Wang ,&nbsp;Liyan Yu ,&nbsp;Lixin Zhang ,&nbsp;Tao Zhang","doi":"10.1016/j.synbio.2024.12.006","DOIUrl":"10.1016/j.synbio.2024.12.006","url":null,"abstract":"<div><div>Genome mining has revealed that <em>Penicillium</em> spp. possess numerous down-regulated or cryptic biosynthetic gene clusters (BGCs). This finding hinted that our investigation of fungal secondary metabolomes is limited. Herein, we report a genetically-modified activation strategy to characterize the spectrum of sesquiterpenoids produced by <em>Penicillium brasilianum</em> CGMCC 3.4402. The cryptic or down-regulated pathways were stimulated by constitutive expression of pathway-specific regulator gene <em>berA</em> responsible for berkeleyacetals biosynthesis from <em>Neosartorya glabra</em>. Chemical analysis of the extracts from the mutant strain <em>Pb</em>-OE:<em>berA</em> enabled the isolation of two new compounds including one bisabolene-type arpenibisabolane C (<strong>1</strong>), one daucane-type arpenicarotane C (<strong>4</strong>), along with four known sesquiterpenoids including arpenibisabolane A (<strong>2</strong>), eupenicisirenins A (<strong>3</strong>), arpenicarotane B (<strong>5</strong>) and aspterric acid (<strong>6</strong>). The assignments of their structures were elucidated from detailed analyses of spectroscopic data, electronic circular dichroism calculation, and biogenetic considerations. The bioassay of isolated compounds (<strong>1</strong>–<strong>6</strong>) exhibited no cytotoxic activities against three tumor cells including MCF-7, HepG2, and A549. Arpenibisabolane C (<strong>1</strong>) and A (<strong>2</strong>) showed weak inhibition bioactivities on aquatic pathogens <em>Vibrio owensii</em> and <em>Vibrio algivorus</em>. Moreover, phylogenetic analysis and sequence alignments of crucial sesquiterpene synthases were performed. Based on the chemical structures and biogenetic investigations, a hypothetic pathway of new compounds (<strong>1</strong>, <strong>4</strong>) was proposed.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 2","pages":"Pages 391-400"},"PeriodicalIF":4.4,"publicationDate":"2024-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11745945/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143011963","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metabolic engineering of Glarea lozoyensis for high-level production of pneumocandin B0","authors":"Xinyi Zhang ,&nbsp;Shu Cheng ,&nbsp;Jing Yang ,&nbsp;Li Lu ,&nbsp;Zixin Deng ,&nbsp;Guangkai Bian ,&nbsp;Tiangang Liu","doi":"10.1016/j.synbio.2024.12.008","DOIUrl":"10.1016/j.synbio.2024.12.008","url":null,"abstract":"<div><div>Pneumocandin B₀ (PB₀) is a lipohexapeptide synthesized by <em>Glarea lozoyensis</em> and serves as the precursor for the widely used antifungal drug caspofungin acetate (Cancidas®). However, the low titer of PB₀ results in fermentation and purification costs during caspofungin production, limiting its widespread clinical application. Here, we engineered an efficient PB₀-producing strain of <em>G. lozoyensis</em> 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 PB₀ 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 PB₀ and offers general guidance for developing efficient fungal cell factories to produce polyketide synthase-non-ribosomal peptide synthetase hybrid metabolites.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 2","pages":"Pages 381-390"},"PeriodicalIF":4.4,"publicationDate":"2024-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11742615/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143011966","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}