Synthetic and Systems Biotechnology最新文献

{"title":"The functional analysis of NirBD in microorganisms and the efficacies of NirBD on the soil nitrogen storage and N2O emission","authors":"Yidan Peng ,&nbsp;Tengxia He ,&nbsp;Qimin Zhou ,&nbsp;Mengyuan Yin ,&nbsp;Chengtao Jin","doi":"10.1016/j.synbio.2025.12.017","DOIUrl":"10.1016/j.synbio.2025.12.017","url":null,"abstract":"<div><div>The siroheme-containing nitrite reductase (NirBD), which is encoded by the <em>nirBD</em> gene, is a functional enzyme in the nitrogen cycle. The NirBD enzyme can regulate N₂O release through the two distinct pathways of assimilatory nitrate/nitrite reduction to biomass and dissimilatory nitrate/nitrite reduction to ammonium. Therefore, a thorough comprehension of the function of NirBD in microorganisms can enable us to better understanding the contributions for nitrogen retention and N₂O emission. However, the knowledge of the functions and expression mechanisms of <em>nirBD</em> gene across different microorganisms remains limited. This review synthesized the current research on the phylogenetic distribution and catalytic versatility of NirBD in fungi, bacteria, and actinomycetes. The contributions of NirBD for nitrogen retention and N₂O emission were extensively discussed under anaerobic and aerobic conditions. The expression mechanism of NirBD was demonstrated. The factors that affect the expression amount of the NirBD enzyme in microorganisms were clarified systematically. This review not only elucidated the unique role of NirBD in the nitrogen metabolism but also provided a critical theoretical foundation for developing future strategies to enhance soil nitrogen fertility and mitigate N₂O emissions.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"12 ","pages":"Pages 364-373"},"PeriodicalIF":4.4,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145975969","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a suite of activatable plant synthetic promoter systems using a bacterial LysR-type transcriptional regulator FdeR","authors":"Yinan Wu ,&nbsp;Curtis Chen ,&nbsp;Sijin Li","doi":"10.1016/j.synbio.2025.12.005","DOIUrl":"10.1016/j.synbio.2025.12.005","url":null,"abstract":"<div><div>Advancing plant synthetic biology requires an abundant supply of orthogonal and tunable genetic parts to express multiple genes simultaneously. Current genetic parts, particularly promoters, used in plants are still limited and often suffer from tissue specificity and endogenous regulation <em>in planta</em>. Synthetic promoter systems that combine engineered plant-compatible transcription factors (TFs) with synthetic promoters provide a promising alternative approach to enrich current plant synthetic biology toolkits. Leveraging the feature that LysR-type TFs usually bind to the operators regardless of the presence or absence of ligands, we present a systematic approach to develop and characterize a large suite of synthetic promoter systems based on a single LysR-type bacterial TF. Using FdeR from the soil bacterium <em>Herbaspirillum seropedicae</em> and its corresponding operator, <em>fdeO</em>, we developed 52 synthetic promoter systems regulated by four FdeR-derived synthetic activators. Transient expression assays in <em>Nicotiana benthamiana</em> showed that the synthetic promoters were constitutively activated by synthetic activators in a ligand-independent manner, and the resulting promoters spanned a wide range of expression activities, with the lowest equivalent to minimal CaMV 35S promoter and the highest with ∼65% of CaMV 35S activity. We also developed modular workflows and pipelines to accelerate the development of the synthetic transcription platform, including a MoClo-based plug-and-play assembly system for plant-compatible promoter construction and a yeast-based prescreening platform for rapid TF evaluation in eukaryotic cells. This work has demonstrated a scalable framework for developing a large set of synthetic promoter systems from minimal genetic components, leveraging bacterial TF diversity to expand the plant synthetic biology toolkit for robust, orthogonal gene expression.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"12 ","pages":"Pages 312-319"},"PeriodicalIF":4.4,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145883697","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Defect-complementation homologous recombination: A novel strategy for precise genome engineering of virulent phages","authors":"Hailin Zhang ,&nbsp;Yueyue Song ,&nbsp;Wenyue Liu ,&nbsp;Xiaoqing Zheng ,&nbsp;Xiaodong An ,&nbsp;Chao Li ,&nbsp;Weihua Chen ,&nbsp;Hailong Wang ,&nbsp;Yuran Zhang","doi":"10.1016/j.synbio.2025.11.002","DOIUrl":"10.1016/j.synbio.2025.11.002","url":null,"abstract":"<div><div>Engineered bacteriophages (phages) have been developed to overcome the limitations of natural phage therapies and serve as precision-targeted agents against drug-resistant bacterial infections. However, their application has been constrained by the low efficiency of existing genome-editing tools, largely because of the absence of effective selection markers. This study proposed a novel strategy, termed <u>d</u>efect-<u>c</u>omplementation <u>h</u>omologous <u>r</u>ecombination (DCHR), for precise phage genome editing. In this approach, CRISPR-Cas9 cleaves a donor plasmid in host cells to release a linear donor template carrying homology arms, an essential phage gene used as a selection marker, and two <em>lox</em> sites. The donor template undergoes homologous recombination with the genome of essential gene-deficient phage, thereby enabling targeted genome modifications. Using DCHR, we successfully generated large genomic deletions (1.48-kb <em>gp0.4–0.7</em> and 1.02-kb <em>gp4.3–4.7</em>), achieved gene insertion (3.08-kb <em>lacZ</em>), and introduced a single-base substitution (T<u>G</u>A to T<u>A</u>A) in the stop codon of <em>gp9</em> within the same T7 phage genome, all with 100 % accuracy. The significant advantages of DCHR are as follows: (i) High-efficiency screening: Only progeny phages derived from successful homologous recombination retain viability and replicative capacity, thereby greatly simplifying recombinant isolation. (ii) Editing flexibility: Unlike CRISPR-Cas systems, DCHR cannot be constrained by protospacer adjacent motif dependence and allows modifications across diverse genomic loci. (iii) High recombination efficiency: DCHR can achieve a recombinant phage titer of 3.1 × 10⁵ PFU mL⁻¹ (plaque-forming units per mL) without relying on exogenous homologous recombination systems. In summary, DCHR demonstrates potential as a precise and efficient general genome-editing tool that facilitates design of engineered phages and advances functional genomic studies.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"12 ","pages":"Pages 59-70"},"PeriodicalIF":4.4,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145577484","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Leveraging ANXA1 to enhance recombinant protein yields in CHO cells: A UPR-Mediated bioprocessing approach","authors":"Qi Zhao ,&nbsp;Hui-Jie Zhang ,&nbsp;Ming-Ming Han ,&nbsp;Jumai Abiti ,&nbsp;Yan-Ju Dong ,&nbsp;Jia-Ning Wang ,&nbsp;Jiang-Tao Lu ,&nbsp;Wen Wang ,&nbsp;Xi Zhang ,&nbsp;Shao-Lei Geng ,&nbsp;Le-Le Qiu ,&nbsp;Xiao-Yin Wang ,&nbsp;Zi-Chun Hua ,&nbsp;Tian-Yun Wang ,&nbsp;Yan-Long Jia","doi":"10.1016/j.synbio.2025.12.001","DOIUrl":"10.1016/j.synbio.2025.12.001","url":null,"abstract":"<div><div>Chinese hamster ovary (CHO) cells undergo endoplasmic reticulum stress (ERS) during intensive recombinant protein production, triggering the unfolded protein response (UPR) to balance cell survival and protein output. Nevertheless, key regulatory components of this process remain incompletely characterized. In this study, we demonstrate that Annexin A1 (ANXA1) functions as a UPR suppressor in CHO cells. Employing the PiggyBac transposon system, we generated a stable ANXA1-knockdown cell line exhibiting a 4.5-fold increase in recombinant antibody expression and a 4.2-fold increase in specific productivity. Pharmacological inhibition using AC2-26 similarly enhanced recombinant protein expression in low-productivity cell populations. Mechanistically, ANXA1 depletion remodeled the UPR by activating the PERK-eIF2α-ATF4 and IRE1-XBP1 branches. This activation upregulaed ATF4, Bip, and XBP1s; suppressed CHOP; reduced apoptosis; and enhanced autophagic flux. Metabolic profiling revealed increased glucose and lactate utilization, while glutamine consumption and ammonia flux remained unchanged. Collectively, these findings establish that ANXA1 depletion enhances recombinant protein biosynthesis through coordinated pro-survival mechanisms. Targeting ANXA1 thus represents an innovative cell engineering strategy for optimizing CHO cell platforms in industrial biopharmaceutical manufacturing.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"12 ","pages":"Pages 197-208"},"PeriodicalIF":4.4,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145797028","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Biosynthesis of patchoulol via metabolic engineering the oleaginous red yeast Rhodotorula toruloides","authors":"Xiaochun Zheng ,&nbsp;Yajun Li ,&nbsp;Zhenhua Liu ,&nbsp;Peng Sun ,&nbsp;Yaqi Dong ,&nbsp;Xue Wang ,&nbsp;Yanan Wang ,&nbsp;Xiaobing Yang","doi":"10.1016/j.synbio.2025.12.003","DOIUrl":"10.1016/j.synbio.2025.12.003","url":null,"abstract":"<div><div>Patchoulol is a widely used sesquiterpenoid in perfumes, cosmetics, foods and pharmaceuticals. The plant-dependent production is suffering from limited growing area, long seasonal cycle, etc. Microbial production represents a sustainable alternative as it is featured with mild operating conditions and eco-friendliness. Herein, we engineered the oleaginous <em>Rhodotorula toruloides</em> toward patchoulol production. First, the patchoulol biosynthesis baseline was constructed by employing a chimeric enzyme of the <em>Pogostemon cablin</em> originated patchoulol synthase and the native FPPS. Second, the supply of essential intermediates was streamlined by redeploying the mevalonate (MVA) pathway while the recycling of NADPH was enhanced through over-expressing related enzymes. Third, the patchoulol production was further enhanced to 724.8 mg/L, 6.0 mg/L/h and 36.2 mg/g glucose by down-regulating the squalene biosynthesis and tuning the cultivation condition in shake flask. Finally, the production of patchoulol was increased to 1.31 g/L and 13.8 mg/g glucose in the minimal medium in a 3-L bioreactor. Our study demonstrated the potential of <em>R. toruloides</em> in producing patchoulol, and should shed light on the microbial synthesis of other sesquiterpenes.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"12 ","pages":"Pages 293-300"},"PeriodicalIF":4.4,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145883698","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Regulating protein glycosylation modification enhances the synthesis of taxadiene in Saccharomyces cerevisiae","authors":"Chenglong Zhang ,&nbsp;Jia Wang ,&nbsp;Longfei Zhao ,&nbsp;Nan Wu ,&nbsp;Yi Shi ,&nbsp;Xia Li ,&nbsp;Changqiang Ke ,&nbsp;Jia Liu ,&nbsp;Yang Ye ,&nbsp;Ying Wang ,&nbsp;Bingzhi Li ,&nbsp;Wenhai Xiao ,&nbsp;Mingdong Yao ,&nbsp;Yingjin Yuan","doi":"10.1016/j.synbio.2025.12.012","DOIUrl":"10.1016/j.synbio.2025.12.012","url":null,"abstract":"<div><div>Utilizing synthetic biology techniques to construct recombinant engineered cells for the synthesis of paclitaxel and its key precursors has emerged as an effective method to address the supply–demand imbalance and protect rare plant resources. Taxadiene, a critical precursor of paclitaxel, exhibits limited yield in eukaryotic systems, constraining its biosynthetic potential. Previous research has demonstrated that glycosylation modifications in <em>Saccharomyces cerevisiae</em> substantially impact the regulation of exogenous protein expression. In this study, we found that knocking out endogenous protein glycosylation genes in the chassis significantly improved the expression of heterologous proteins, especially the key taxadiene synthase (TS), and thereby increased the yield of taxadiene. In particular, we identified that the deletion of the glycosyltransferase gene <em>mnn11</em> increased taxadiene production levels by 65.2 %. The subsequent multi-copy integration of the key enzyme taxadiene synthase further elevated taxadiene production levels in shake flasks by more than 3-fold, reaching 113.5 mg/L. Moreover, the enhancement of geranylgeranyl diphosphate synthesis-related expression modules resulted in a 2.69-fold increase in taxadiene yield, to 420.4 mg/L. Following the optimization of fed-batch fermentation conditions, taxadiene production levels of up to 1.26 g/L were achieved, representing a 63-fold improvement over that obtained with the initial strain. Our findings provide valuable insights into enhancing heterologous taxadiene production efficiency by blocking endogenous protein glycosylation modifications in <em>S. cerevisiae</em>, establishing a critical precedent for improving compatibility between natural product synthesis and microbial cell factories.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"12 ","pages":"Pages 284-292"},"PeriodicalIF":4.4,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145883633","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"SPIN-guided engineering of a novel (R)-amine transaminase from Fusarium albosuccineum for enantioselective synthesis of chiral piperidyl amines","authors":"Ruizhou Tang ,&nbsp;Jiahuan Li ,&nbsp;Xiaole Yang ,&nbsp;Xia Tian ,&nbsp;Zehui Wang ,&nbsp;Xuning Zhang ,&nbsp;Tingting Li","doi":"10.1016/j.synbio.2025.12.010","DOIUrl":"10.1016/j.synbio.2025.12.010","url":null,"abstract":"<div><div>Aminotransferases are promising green biocatalysts for the synthesis of chiral amines, yet their limited catalytic efficiencies restrict broader industrial applications. In this study, a novel (<em>R</em>)-amine transaminase, FalAT, was identified from <em>Fusarium albosuccineum</em> through genome mining. FalAT exhibited optimal activity at 30 °C and pH 7.0 and catalyzed the conversion of 1-Boc-3-piperidone to (<em>R</em>)-1-Boc-3-aminopiperidine with &gt;99 % enantiomeric excess. To efficiently enhance its catalytic performance, a Substrate–Protein Interaction Network (SPIN) strategy was implemented, integrating structure-guided analysis, molecular docking, virtual saturation mutagenesis, and dual energy–distance filtering. SPIN first screened and constructed a mutational library covering 70 amino acid positions, which was subsequently narrowed to 9 key residues, ultimately yielding 15 candidate mutants for experimental validation. Experimental results showed that five mutants exhibited higher catalytic activity than the wild-type enzyme, among which R126A was the most effective, displaying approximately a 4-fold increase in catalytic activity and a 13-fold enhancement in catalytic efficiency (<em>k</em>_<em>ca</em><em>_t/K</em>_<em>m</em> = 2.05 s⁻¹ mM⁻¹). Molecular dynamics simulations revealed that the R126A mutation expanded the active-site cavity, alleviated steric hindrance, and strengthened hydrophobic interactions, thereby improving substrate binding and catalytic turnover. Furthermore, substrate profiling demonstrated that FalAT possesses moderate substrate promiscuity. Overall, the SPIN strategy significantly improved the catalytic performance of FalAT while markedly reducing experimental workload, providing an efficient and generalizable approach for the directed evolution of (<em>R</em>)-amine transaminases for the green synthesis of chiral amines.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"12 ","pages":"Pages 229-237"},"PeriodicalIF":4.4,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145883628","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Efficient whole-cell biocatalytic synthesis of 2′-deoxy-2′-fluoroadenosine, a key building block for nucleic acid drugs","authors":"Dandan Feng ,&nbsp;Tianshu Zhang ,&nbsp;Guan Zhou ,&nbsp;Yanli Cao ,&nbsp;Quan Luo ,&nbsp;Huifang Xu ,&nbsp;Xuefeng Lu","doi":"10.1016/j.synbio.2025.11.014","DOIUrl":"10.1016/j.synbio.2025.11.014","url":null,"abstract":"<div><div>2′-Deoxy-2′-fluoroadenosine (2′-F-dA) is a nucleoside analogue used as a key building block for oligonucleotide drugs. It can be biosynthesized from a low-cost 2′-deoxy-2′-fluorouridine via one-pot transglycosylation catalyzed by a thymidine phosphorylase (TP) and a purine nucleoside phosphorylase (PNP). However, reliance on purified enzymes and low space-time yields present challenges for industrial application of the process. Here, we develop a whole-cell-based biocatalytic system employing TP and PNP from <em>Escherichia coli</em>, which demonstrates high catalytic efficiency and operational simplicity in scaled-up reaction. In particular, a thermal pretreatment of TP- and PNP-expressing whole cells, determined as 50 °C for 3 h, effectively suppressed endogenous deamination side reaction while enhancing 2′-F-dA yield. Subsequent optimization of enzyme and substrate loadings and their relative ratios achieved an unprecedented space-time yield of 1.22 g/L/h with 88.1 g/L product titer in a 500 mL scaled-up reaction, manifesting a highest total conversion of 68.2 %. An integrated purification process yielded gram-scale solid powder of 2′-F-dA with 98.0 % chemical purity and 85.0 % recovery. This novel whole-cell biocatalytic process demonstrates significant industrial potential for the production of 2′-F-dA.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"12 ","pages":"Pages 152-159"},"PeriodicalIF":4.4,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145690537","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluation of synthetic post-transcription regulatory sequences reveals design principle to enhance mRNA stability and translation efficiency","authors":"Jin-Peng Zhang ,&nbsp;Zi-Lun Mei ,&nbsp;Jia-Wei Ren ,&nbsp;Xiao-Mei Zhang ,&nbsp;Jin-Song Gong ,&nbsp;Guo-Qiang Xu ,&nbsp;Hui Li ,&nbsp;Xiao-Juan Zhang ,&nbsp;Zheng-Hong Xu","doi":"10.1016/j.synbio.2025.11.011","DOIUrl":"10.1016/j.synbio.2025.11.011","url":null,"abstract":"<div><div>The Shine-Dalgarno (SD) sequence and its adjacent flanking regions, including the translation standby site (TSS) and the N-terminal coding sequence (NCS), play critical roles in regulating ribosome recruitment, translation efficiency, and mRNA stability against RNase degradation. However, structure-activity relationships governing these regions remain poorly characterized, and their functional interplay introduces substantial complexity. In this study, we employed one-pot technology to build a post-transcriptional regulatory component (PTRC) library of 576 variants to clarify the relationship between sequence variants and both protein expression and mRNA levels via high-throughput sequencing. Our results show that although unstructured TSSs do not enhance mRNA stability, they markedly increase translation efficiency, causing a 16 %–100 % rise in protein expression. In contrast, structured TSSs increase mRNA levels by 43 %–90 %. Additionally, highly conserved SD sequences boost translation efficiency by up to 10 % and mRNA abundance by up to 12 %. Moreover, it was found that optimized linear N-terminal coding sequence (NCS) positively affects protein expression and mRNA levels. The effects of these optimized regulatory components were verified in the expression control of the <em>nrk</em> and <em>sam2</em> genes, resulting in enhanced production. These findings underscore the crucial role of structural optimization, guiding the rational design of synthetic post-transcriptional regulatory elements.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"12 ","pages":"Pages 160-171"},"PeriodicalIF":4.4,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145747942","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineering Yarrowia lipolytica for high-level ergosterol production","authors":"Xinyu Liu ,&nbsp;Qihang Chen ,&nbsp;Wenbao Zhao ,&nbsp;Qi Li ,&nbsp;Liushuting Xiao ,&nbsp;Qian He ,&nbsp;Weizhu Zeng ,&nbsp;Jingwen Zhou","doi":"10.1016/j.synbio.2026.01.001","DOIUrl":"10.1016/j.synbio.2026.01.001","url":null,"abstract":"<div><div>Ergosterol is the key precursor of steroid drug synthesis. In this experiment, we systematically modified the synthesis of ergosterol. Firstly, we identified key rate-limiting enzymes through systematic screening of the post-squalene pathway. Combinatorial overexpression of <em>IDI1</em>, <em>tHMG1</em>, <em>ERG4</em>, <em>ERG5</em>, <em>ERG27</em>, <em>ERG1</em> and <em>ERG11</em> achieved an ergosterol titer of 94.2 mg/L. Molecular dynamics guided mutagenesis of key substrate channel residues, particularly S372V Erg11, enhanced local flexibility and significantly increased ergosterol production. Introduction of the proton-donating mutations S372V-T305H-<em>ERG11</em> established an artificial proton-dependent pathway, which, together with channel engineering, further increased the titer to 124 mg/L. Lipid droplet engineering and cellular compartmentalization strategies increased the titer to 148.3 mg/L. Ultimately, multi-copy integration of all ergosterol synthesis pathway genes increased the titer to 433.1 mg/L, and fed-batch fermentation in a 5-L bioreactor resulted in a final titer of 4.58 g/L. This study demonstrates a comprehensive hierarchical strategy for high-level sterol production.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"12 ","pages":"Pages 393-400"},"PeriodicalIF":4.4,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145975972","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}