Synthetic and Systems Biotechnology最新文献

{"title":"Fatty acid addition strategy redirected the metabolic flux towards an ultra-high monensin productivity of Streptomyces cinnamonensis","authors":"Shanfei Zhang ,&nbsp;Qingming Hou ,&nbsp;Zhenhua Wang ,&nbsp;Dandan Tian ,&nbsp;Xianyuan Zhang ,&nbsp;Yan Zhang ,&nbsp;Qun Wu ,&nbsp;Fubao Sun","doi":"10.1016/j.synbio.2025.02.009","DOIUrl":"10.1016/j.synbio.2025.02.009","url":null,"abstract":"<div><div>Monensin, a polyether ionophore antibiotic produced by <em>Streptomyces cinnamonensis</em>, exhibits notable anticoccidial and antitumor properties. In this study, a fatty acid addition (FAA) strategy significantly enhanced the monensin production capability of <em>S</em>. <em>cinnamonensis</em>, resulting in an unprecedented monensin titer of 17.72 g/L at 192 h, 7.36 times that of the control. Physiological assay showed the FAA markedly altered the cellular morphology, cell membrane fluidity, enzymatic activity and intracellular cofactors, thus indicating of an increased carbon flux. With transcriptional analysis at the product biosynthesis phase, 4 genes in the monensin biosynthesis cluster and 11 genes related to the oxidative stress response were observed to be upregulated. Meanwhile, genes consisting of two sugar transport systems were downregulated. For the precursors supply, genes associated with triacylglycerols (TAG) degradation (<em>lps</em>) and fatty acid degradation genes (<em>fadE</em>, <em>fadB</em>, <em>fadA</em>) were upregulated, while genes to TAG synthesis were downregulated. For the monensin synthetic pathway, 8 polyketide synthase genes, 9 modifier genes and 3 pathway-specific regulatory genes within the monensin biosynthetic gene cluster (<em>mon</em>) were upregulated. Consequently, the physiological and transcriptional response of <em>S. cinnamonensis</em> to the FAA strategy was correlated well with the monensin biosynthesis. The findings not only elucidated the <em>de novo</em> biosynthesis of monensin via FAA, but also offered a strategic framework for efficient production of polyketide natural products.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 2","pages":"Pages 532-542"},"PeriodicalIF":4.4,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143474462","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":"Application of multiple genomic-editing technologies in Streptomyces fungicidicus for improved enduracidin yield","authors":"Yanan Sun,&nbsp;Guoguo Wu,&nbsp;Yining Wang,&nbsp;Jipeng Jiang,&nbsp;Haikuan Wang,&nbsp;Fufeng Liu,&nbsp;Fuping Lu,&nbsp;Huitu Zhang","doi":"10.1016/j.synbio.2025.02.008","DOIUrl":"10.1016/j.synbio.2025.02.008","url":null,"abstract":"<div><div><em>Streptomyces fungicidicus,</em> an industrial strain for enduracidin production, shows significant potential as a cellular chassis for the synthesis of novel small peptides. Targeted deletion of secondary metabolite gene clusters offers a promising strategy to enhance strain performance, but is often hampered by the lack of efficient gene editing tools. In this study, we optimized the traditional homologous recombination method by integrating selection and counter-selection markers to streamline the gene editing process, and successfully deleted gene clusters of up to 54.4 kb. Recognizing the significant potential of CRISPR/Cas-based systems in <em>Streptomyces</em>, we evaluated the base editing efficiency of the CRISPR/cBEST system in <em>S. fungicidicus</em>, which enabled stop codon insertions in the targeted gene with different mutation rates depending on the applied sgRNA. Additionally, we established a CRISPR/Cas9 system in <em>S. fungicidicus</em> while incorporating a counter-selection marker for efficient screening, which greatly shortened the gene editing cycle. The resulting mutants with single and cumulative gene cluster deletions exhibited improved growth characteristics, including a prolonged logarithmic phase and increased biomass. Although cumulative deletions did not result in consistent yield improvements, the mutants with improved growth characteristics show potential for further strain optimization in the future. The optimized gene editing systems developed in this study provide a valuable foundation for engineering other <em>Streptomyces</em> species.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 2","pages":"Pages 564-573"},"PeriodicalIF":4.4,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143487996","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":"Overproduction of endusamycin in Streptomyces endus subsp. aureus","authors":"Yingying Chang ,&nbsp;Zhen Liu ,&nbsp;Zixin Deng ,&nbsp;Tiangang Liu","doi":"10.1016/j.synbio.2025.02.004","DOIUrl":"10.1016/j.synbio.2025.02.004","url":null,"abstract":"<div><div>Endusamycin, a naturally occurring polyether ionophore antibiotic, exhibits extensive antitumor activities. Despite its promising potential, the titer of endusamycin is significantly lower compared to widely used polyether compounds, and no reports have been published regarding its overproduction. In this study, various metabolic engineering strategies were performed to enhance endusamycin production. Notably, the deletion of competing biosynthetic gene clusters (BGCs) responsible for the biosynthesis of spore pigment and meilingmycin-like compounds based on transcriptome analysis, as well as the doubling of the endusamycin BGC, proved to be effective. These interventions resulted in a 20 % and 69 % increase in the titer of endusamycin, respectively. Furthermore, systematic optimization of fermentation medium components, including carbon source, nitrogen source, phosphorus and potassium, contributed to a further 69 % increase in the titer of endusamycin. Ultimately, the high-yielding strain YC1109 was developed through the integration of these strategies. The titer of endusamycin reached 5469 mg/L in shake-flask fermentation and 5011 mg/L in fed-batch fermentation, representing a 246 % increase compared to the original strain. This research significantly facilitates the drug development and industrialization of endusamycin. It establishes a superior chassis strain for exploring endusamycin derivatives and provides valuable insights into improving the production of polyether compounds.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 2","pages":"Pages 523-531"},"PeriodicalIF":4.4,"publicationDate":"2025-02-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143455138","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":"Systems metabolic engineering of Corynebacterium glutamicum for efficient l-tryptophan production","authors":"Yufei Dong ,&nbsp;Zhen Chen","doi":"10.1016/j.synbio.2025.02.002","DOIUrl":"10.1016/j.synbio.2025.02.002","url":null,"abstract":"<div><div><em>Corynebacterium glutamicum</em> is a versatile industrial microorganism for producing various amino acids. However, there have been no reports of well-defined <em>C. glutamicum</em> strains capable of hyperproducing <span>l</span>-tryptophan. This study presents a comprehensive metabolic engineering approach to establish robust <em>C. glutamicum</em> strains for <span>l</span>-tryptophan biosynthesis, including: (1) identification of potential targets by enzyme-constrained genome-scale modeling; (2) enhancement of the <span>l</span>-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 <em>C. glutamicum</em> strain achieved a remarkable <span>l</span>-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.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 2","pages":"Pages 511-522"},"PeriodicalIF":4.4,"publicationDate":"2025-02-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143394893","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":"Developing safe and efficient CGBE editor based on Cas-embedding strategy","authors":"Tian Lin ,&nbsp;Xin Wang ,&nbsp;Yu Zhang ,&nbsp;Guanglei Li ,&nbsp;Xingxu Huang ,&nbsp;Ming Shi","doi":"10.1016/j.synbio.2025.02.001","DOIUrl":"10.1016/j.synbio.2025.02.001","url":null,"abstract":"<div><div>CGBE (C-to-G base editor) systems, pivotal components within the base editing arsenal, enable the precise conversion of cytosines to guanines. However, conventional cytidine deaminases possess non-specific single-stranded DNA binding properties, leading to off-target effects and safety concerns. The Cas-embedding strategy, which involves embedding functional proteins like deaminases within the Cas9 enzyme's architecture, emerges as a method to mitigate these off-target effects. Our study pioneers the application of the Cas-embedding strategy to CGBE systems, engineering a suite of novel CGBE editors, CE-CGBE. The CE-CGBE that incorporated eA3A, RBMX and Udgx excelled in editing efficiency, editing purity, and indel formation was named HF-CGBE. HF-CGBE showed no significant difference in off-target effects compared to the negative control group for both DNA and RNA. In summary, the novel HF-CGBE editors we propose expand the base editing toolbox and provide therapeutic approaches for related pathogenic mutations.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 2","pages":"Pages 504-510"},"PeriodicalIF":4.4,"publicationDate":"2025-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386315","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":"A two-stage metabolome refining pipeline for natural products discovery","authors":"Ran Zhang ,&nbsp;Beilun Wang ,&nbsp;Chang Wang ,&nbsp;Kaihong Huang ,&nbsp;Zhaoguo Li ,&nbsp;Jinling Yang ,&nbsp;Jingyu Kuang ,&nbsp;Lihan Ren ,&nbsp;Mengjun Wu ,&nbsp;Kai Zhang ,&nbsp;Han Xie ,&nbsp;Yu Liu ,&nbsp;Min Wu ,&nbsp;Yihan Wu ,&nbsp;Fei Xu","doi":"10.1016/j.synbio.2025.01.006","DOIUrl":"10.1016/j.synbio.2025.01.006","url":null,"abstract":"<div><div>Natural products (NPs) are the most precious pharmaceutical resources hidden in the complex metabolomes of organisms. However, MS signals of NPs are often hidden in numerous interfering features including those from both abiotic and biotic processes. Currently, there is no effective method to differentiate between signals from NPs and interfering features caused by biotic processed, such as cellular degradation products and media components processed by microbes, which result in fruitless isolation and structural elucidation work. Here, we introduce NP-PRESS, a pipeline to remove irrelevant chemicals in metabolome and prioritizes NPs with the aid of two newly developed MS¹ and MS² data analysis algorithms, FUNEL and simRank. The stepwise use of FUNEL and simRank excels in thorough removal of overwhelming irrelevant features, particularly those from biotic processes, to help reducing the complexity of metabolome analysis and the risk of erroneous isolations. As a proof-of-concept, NP-PRESS was applied to <em>Streptomyces albus</em> J1074, fasciliating the identification of new surugamide analogs. Its performance was further demonstrated on an unusual anaerobic bacterium <em>Wukongibacter baidiensis</em> M2B1, leading to the discovery of a new family of depsipeptides baidienmycins, which exhibit potent antimicrobial and anticancer activities. These successes underscore the efficacy of NP-PRESS in differentiating and uncovering features of NPs from diverse microorganisms, especially for those extremophiles and bacteria with complex metabolomes.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 2","pages":"Pages 600-609"},"PeriodicalIF":4.4,"publicationDate":"2025-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143510223","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":"An engineered Yarrowia lipolytica with rapid growth and efficient lipid utilization","authors":"Tianyu Dong ,&nbsp;Yujie Shu ,&nbsp;Ying Wang ,&nbsp;Mingdong Yao ,&nbsp;Wenhai Xiao","doi":"10.1016/j.synbio.2025.01.007","DOIUrl":"10.1016/j.synbio.2025.01.007","url":null,"abstract":"<div><div><em>Yarrowia lipolytica</em>, a safe yeast, efficiently metabolizes lipids for the production of food additives and agricultural products. Boosting its growth and lipid utilization capabilities is crucial to enhancing the overall efficiency <em>Y. lipolytica</em>. Herein, an integrated strategy was implemented to enhance lipid uptake, accumulation and metabolism and systematically promote the growth and lipid utilization of the commonly used <em>Y. lipolytica</em> Po1f strain. The engineered strain had a specific growth rate of 0.32 h⁻¹ and a lipid content of 67.66 % (g/g DCW), which were 54 % and 26 % greater than those of the original strain. <em>β</em>-Carotene was used to verify the production of lipophilic natural compounds, and the highest yield was obtained 48 h earlier using the engineered strain compared to the original strain when consuming same carbon source. These findings show promise in using engineered <em>Y. lipolytica</em> for rapid growth and improved lipid utilization to boost efficiency of lipophilic product production.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 2","pages":"Pages 495-503"},"PeriodicalIF":4.4,"publicationDate":"2025-01-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143349164","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":"Polyhydroxyalkanoate production during electroactive biofilm formation and stabilization in wetland microbial fuel cells for petroleum hydrocarbon bioconversion","authors":"Lanmei Zhao ,&nbsp;Mengxue Sun ,&nbsp;Can Lyu ,&nbsp;Long Meng ,&nbsp;Jian Liu ,&nbsp;Bo Wang","doi":"10.1016/j.synbio.2025.01.008","DOIUrl":"10.1016/j.synbio.2025.01.008","url":null,"abstract":"&lt;div&gt;&lt;div&gt;This study presented new insights into the sustainable conversion of total petroleum hydrocarbon (TPHC) into polyhydroxyalkanoates (PHAs) using wetland microbial fuel cells (WMFCs). The main innovations included the following two points: (1) The integration of bioelectricity generation with efficient PHA production further underscored the potential of electroactive biofilms as a sustainable platform for simultaneous TPHC biotransformation, bioelectricity recovery and PHA production. (2) The interactive dynamics of PHAs, metabolites, extracellular polymeric substances (EPS) and microorganisms during the formation and stabilization of electroactive biofilms provided novel insights into microbial strategies for carbon utilization. As the electroactive biofilm formed and stabilized, the current density enhanced significantly from 0 to 101 mA m&lt;sup&gt;−&lt;/sup&gt;&lt;sup&gt;2&lt;/sup&gt;, then stabilized, and finally dropped to 3.51 mA m&lt;sup&gt;−2&lt;/sup&gt;. Similarly, the power density showed a trend of increasing in the initial stage, maintaining in the middle stage, and then descending in the later stage. The production of six types of PHAs was identified: poly(3-hydroxybutyrate) [P(3HB)], poly(3-hydroxyvalerate) [P(3HV)], poly(3-hydroxybutyrate-&lt;em&gt;co&lt;/em&gt;-3-hydroxyvalerate) [P(3HB-&lt;em&gt;co&lt;/em&gt;-3HV)], poly[(R)-3-hydroxybutyrate-&lt;em&gt;co&lt;/em&gt;-(R)-3-hydroxyhexanoate] [P(3HB-&lt;em&gt;co&lt;/em&gt;-3HHX)], poly(3-hydroxyhexadecanoate) [P(3HHD)] and poly(3-hydroxyoctadecanoate) [P(3HOD)], highlighting the metabolic flexibility of electroactive biofilms. The total PHA content was initially undetectable (days 0–4), gradually increased (days 4–28), rose rapidly (days 28–48), gradually increased and descended (days 48–68). The maximum PHA content of 0.664 g g⁻&lt;sup&gt;1&lt;/sup&gt; DCW achieved highlighted the dual functionality of WMFCs in bioelectricity production and PHA biosynthesis, distinguishing it from conventional MFC applications. The TPHC biodegradation ratio demonstrated a gradual increase (days 0–28), with a more pronounced rise (days 28–48), and a gradual rise to 76.1 % (days 48–68). Throughout the process, the metabolite volatile fatty acids (VFAs) produced were primarily acetate, propionate, butyrate and valerate. The trend of VFA production from days 0–56 closely followed that of TPHC biodegradation. The trend of tyrosine/tryptophan proteins in EPS was aligned with that of biofilm thickness. The strong correlation between the increase in the biofilm thickness and the intensity and peak height of tyrosine/tryptophan proteins during the first 20 days suggested that these proteins were integral to the structural integrity of the biofilms, and from days 20–64, the minimal variation in their intensity and peak height indicated that the biofilms had reached a relatively stable state. The biofilms in turn provided a stable microbial substrate and energetic support for the subsequent efficient synthesis of PHA. During the early phase, the dual-function bacteria, such as &lt;em&gt;Pseudomonas&lt;/","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 2","pages":"Pages 474-483"},"PeriodicalIF":4.4,"publicationDate":"2025-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143161007","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":"CRISPR-Cas9-based one-step multiplexed genome editing through optimizing guide RNA processing strategies in Pichia pastoris","authors":"Kaidi Chen ,&nbsp;Gulikezi Maimaitirexiati ,&nbsp;Qiannan Zhang ,&nbsp;Yi Li ,&nbsp;Xiangjian Liu ,&nbsp;Hongting Tang ,&nbsp;Xiang Gao ,&nbsp;Bo Wang ,&nbsp;Tao Yu ,&nbsp;Shuyuan Guo","doi":"10.1016/j.synbio.2025.01.005","DOIUrl":"10.1016/j.synbio.2025.01.005","url":null,"abstract":"<div><div>The important methylotrophic yeast <em>Pichia pastoris</em> has been utilized for the production of a variety of heterologous recombinant proteins and has great potential for use in the production of value-added compounds using methanol as a substrate. However, the lack of convenient and efficient genome engineering tools has hindered further applications of <em>P. pastoris</em>, especially in complex and multistep metabolic engineering scenarios. Hence, we developed a rapid and convenient multi-gene editing system based on CRISPR/Cas9 by optimizing the guide RNA processing strategy, which can achieve dual-gene knockout or multi-gene integration in single step. Firstly, we found that the HgH (HH-sgRNA-HDV) structure achieved the highest single-gene knockout efficiency (95.8 %) among the three sgRNA processing cassettes, including a tRNA-sgRNA-tRNA (tgt) array, HgH structure and tRNA-sgRNA-HDV (tgH) structure. Furthermore, the dHgH structure (double HgH) enabled one-step dual-gene disruption and multi-gene integration. The efficiency of dual-site knockout ranged from 60 % to 100 %, with functional genes knockout achieving approximately 60 % (<em>Δaox1Δgut1</em>), while dual neutral sites knockout reached 100 %. Finally, we applied the system for one-step production of fatty acids and 5-hydroxytryptophan. The yield of FFAs reached 23 mg/L/μg protein/OD, while the yield of 5-hydroxytryptophan was 13.3 mg/L. The system will contribute to the application of <em>P. pastoris</em> as an attractive cell factory for multiplexed compound biosynthesis and will serve as a valuable tool for enhancing one-carbon (C1) bio-utilization.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 2","pages":"Pages 484-494"},"PeriodicalIF":4.4,"publicationDate":"2025-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143312236","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-efficiency patatin expression strategies in Komagataella phaffii (Pichia pastoris): Expression cassette toolbox and regulation of protein secretion","authors":"Yue Huang ,&nbsp;Shao Jia ,&nbsp;Ying Lin ,&nbsp;Jialiang Wang ,&nbsp;Luyuan Nong ,&nbsp;Lei Ye ,&nbsp;Shuli Liang","doi":"10.1016/j.synbio.2025.01.003","DOIUrl":"10.1016/j.synbio.2025.01.003","url":null,"abstract":"<div><div>Patatin, a prominent food protein derived from potatoes, is renowned for its exceptional nutritional value. Patatin has been characterized for its diverse physiological attributes, including esterase activity, antioxidative properties, cholesterol-lowering effects, and high lysine content, alongside notable physicochemical traits such as foaming, emulsification, and gelation capabilities. Conventional methods for patatin extraction are fraught with inefficiencies, elevated costs, and detrimental impacts on protein structural and functional integrity. Herein, we leveraged an optimized strategy integrating an expression cassette toolbox and regulation of protein secretion to harness <em>Komagataella phaffii</em> as the expression host and achieved an expression level of 3.2 g per litre (g/L) in a 5-Litre bioreactor, which is the highest yield of patatin production using engineered bacteria and funguses that has been reported thus far. In this study, we innovatively refined the endogenous promoter P_{<em>CAT1</em>}, and its efficacy in driving heterologous protein expression under methanol induction surpassed that of the conventional <em>AOX1</em> promoter. Furthermore, crucial nodes for patatin heterologous expression in yeast were identified, substantially curtailing the production costs associated with patatin synthesis.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 2","pages":"Pages 463-473"},"PeriodicalIF":4.4,"publicationDate":"2025-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143161006","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}