Journal of biotechnology最新文献_第2页

Efficient biosynthesis of ectoine in engineered Vibrio natriegens 异托因在工程弧菌营养菌中的高效生物合成

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-06-03 DOI: 10.1016/j.jbiotec.2025.06.001

Tianzhi Fang , Mengkai Hu , Dandan Li , Kun Liu , Yu Chen , Zhenglian Xue , Qingtao Liu , Tianwen Wang

{"title":"Efficient biosynthesis of ectoine in engineered Vibrio natriegens","authors":"Tianzhi Fang , Mengkai Hu , Dandan Li , Kun Liu , Yu Chen , Zhenglian Xue , Qingtao Liu , Tianwen Wang","doi":"10.1016/j.jbiotec.2025.06.001","DOIUrl":"10.1016/j.jbiotec.2025.06.001","url":null,"abstract":"<div><div>The compatible solute ectoine, an amino acid derivative, was initially discovered to be a critical factor conferring high resistance to harsh environments in which the producer microbes subsist. Its diverse chemical and biomedical properties have made ectoine a star molecule, necessitating environmentally friendly and efficient preparation methods. In this study, we cloned a biosynthetic gene cluster from a strain screened from samples collected from a historic sea-salt plant. This gene cluster supports inducible heterogeneous ectoine synthesis controlled by the T7 promoter in the recombinant halophile host <em>Vibrio natriegens</em> Vmax × 2. Investigation of the dosage effect of each gene in the cluster on ectoine synthesis identified <em>ectC</em> as the most important gene. We performed <em>in silico</em> directed evolution and Pythia analysis to design <em>ectC</em> mutants and screen those that positively contribute to ectoine synthesis. We used a sequence-generating tool to design degenerated EctC coding sequences to ensure EctC levels in recombinant <em>V. natriegens</em>. Ectoine synthesis was validated in a 5-L stirring fermenter. A titer of 56.47 g/L ectoine was achieved in 28 h through feed batch fermentation. This study demonstrated the power of protein language model-guided metabolic enzyme design and codon adaptation index-based coding sequence selection in biosynthesis pathway engineering. It also offers an example of ectoine biosynthesis in the rising star halophile host <em>V. natriegens</em>, contributing significantly to microbiology and biotechnology.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 238-248"},"PeriodicalIF":4.1,"publicationDate":"2025-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144231662","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}

引用次数: 0

Enabling quantification of 2′-fucosyllactose via ligand-dependent thermal stabilization of BoGT6a 通过BoGT6a的配体依赖热稳定来定量2'-聚焦基乳糖。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-06-03 DOI: 10.1016/j.jbiotec.2025.06.002

Nayeon Kim , Jonghyeok Shin , Jun-Seob Kim , Dae-Hyuk Kweon

{"title":"Enabling quantification of 2′-fucosyllactose via ligand-dependent thermal stabilization of BoGT6a","authors":"Nayeon Kim , Jonghyeok Shin , Jun-Seob Kim , Dae-Hyuk Kweon","doi":"10.1016/j.jbiotec.2025.06.002","DOIUrl":"10.1016/j.jbiotec.2025.06.002","url":null,"abstract":"<div><div>2′-Fucosyllactose (2′-FL) is a major component of Human Milk Oligosaccharides (HMOs) that plays a crucial role in developing the neonatal immune system and modulating gut microbiota. Due to its health benefits, 2′-FL has gained industrial importance as a key ingredient in probiotic products and functional foods. Although quantifying 2′-FL is crucial for its economical production and nutritional management, conventional methods require expensive equipment and skilled personnel, making high-throughput quantification challenging. In this study, we present a simple and cost-effective method for 2′-FL quantification by utilizing the thermal stability of BoGT6a, a glycosyltransferase derived from <em>Bacteroides ovatus</em> that specifically binds to 2′-FL. Initially, the binding of BoGT6a and 2′-FL was confirmed, and we demonstrated that 2′-FL-bound BoGT6a is protected from thermal stress. To achieve rapid detection of 2′-FL, we fused BoGT6a with the fluorescent protein mCherry, resulting in mCherry-BoGT6a, and investigated its thermal stability and fluorescence in response to varying 2′-FL concentrations. Finally, we developed a 2′-FL quantification device that measures protein precipitation with the change of electrical voltage. These results demonstrate the reliability and industrial applicability of BoGT6a-based 2′-FL quantification technology.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 283-289"},"PeriodicalIF":4.1,"publicationDate":"2025-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144234212","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}

引用次数: 0

De novo production of prenylnaringenin compounds by a metabolically engineered Escherichia coli 利用代谢工程大肠杆菌重新生产烯丙基柚皮素化合物。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-05-29 DOI: 10.1016/j.jbiotec.2025.05.017

Daniela Gomes , Joana L. Rodrigues , Nigel S. Scrutton , Ligia R. Rodrigues

{"title":"De novo production of prenylnaringenin compounds by a metabolically engineered Escherichia coli","authors":"Daniela Gomes , Joana L. Rodrigues , Nigel S. Scrutton , Ligia R. Rodrigues","doi":"10.1016/j.jbiotec.2025.05.017","DOIUrl":"10.1016/j.jbiotec.2025.05.017","url":null,"abstract":"<div><div>Prenylnaringenin (PN) compounds, namely 8-prenylnaringenin (8-PN), 3’-prenylnaringenin (3’-PN), and 6-prenylnaringenin (6-PN), are reported to have several interesting bioactivities. This study aimed to validate a biosynthetic pathway for <em>de novo</em> production of PN in <em>Escherichia coli</em>. A previously optimized <em>E. coli</em> chassis capable of efficiently <em>de novo</em> producing naringenin was used to evaluate eleven prenyltransferases (PTs) for the production of PN compounds. As PT reaction requires dimethylallyl pyrophosphate (DMAPP) as extended substrate that has limited availability inside the cells, clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 12a (Cas12a) (CRISPR-Cas12a) was used to construct ten boosted DMAPP-E<em>. coli</em> strains. All the PTs, in combination with the naringenin biosynthetic pathway, were tested in these strains. Experiments in 96-well deep well plates identified twelve strains capable of producing PN. <em>E. coli</em> M-PAR-121 with the integration of the 1-deoxy-D-xylulose-5-phosphate synthase (DXS) gene from <em>E. coli</em> (<em>Ec</em>DXS) into the <em>lacZ</em> locus of the genome (<em>E. coli</em> M-PAR-121:<em>Ec</em>DXS) expressing the soluble aromatic PT from <em>Streptomyces roseochromogenes</em> (CloQ) and the naringenin biosynthetic pathway was selected as the best producer strain. After optimizing the production media in shake flasks, 160.57 µM of 3’-PN, 4.4 µM of 6-PN, and 2.66 µM of 8-PN were obtained. The production was then evaluated at the bioreactor scale and 397.57 µM of 3’-PN (135.33 mg/L) and 25.61 µM of 6-PN (8.72 mg/L) were obtained. To the best of our knowledge, this work represents the first report of <em>de novo</em> production of PN compounds using <em>E. coli</em> as a chassis.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 215-228"},"PeriodicalIF":4.1,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144191777","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}

引用次数: 0

Substitution of reactive centre loop residues from C1 esterase inhibitor increases the inhibitory specificity of alpha-1 antitrypsin for plasma kallikrein C1酯酶抑制剂的反应性中心环残基的替代增加了α -1抗胰蛋白酶对血浆钾激肽的抑制特异性

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-05-28 DOI: 10.1016/j.jbiotec.2025.05.013

Sangavi Sivananthan , S. Ameer Ahmed , Ammaar M. Baig , Varsha Bhakta , William P. Sheffield

{"title":"Substitution of reactive centre loop residues from C1 esterase inhibitor increases the inhibitory specificity of alpha-1 antitrypsin for plasma kallikrein","authors":"Sangavi Sivananthan , S. Ameer Ahmed , Ammaar M. Baig , Varsha Bhakta , William P. Sheffield","doi":"10.1016/j.jbiotec.2025.05.013","DOIUrl":"10.1016/j.jbiotec.2025.05.013","url":null,"abstract":"<div><div>C1 esterase inhibitor (C1INH) is a member of the serpin superfamily of proteins and controls plasma kallikrein (Pka). Purified C1INH concentrates are effective in controlling C1INH deficiency (hereditary angioedema, HAE). Because C1INH is a relatively slow inhibitor of Pka, we sought to develop a more effective inhibitor by exchanging reactive centre loop (RCL) residues in another serpin, alpha-1 antitrypsin (AAT) variant M358R, with the corresponding residues of C1INH. Novel, soluble, N-terminally hexahistidine-tagged variants were expressed in <em>E. coli</em>, purified by nickel chelate chromatography, and characterized kinetically. <strong>A</strong>AT/<strong>C</strong>1INH loop exchange mutants were designated by the RCL residues exchanged using the reactive centre P1-P1′ convention. Maximal exchange mutant AC (10–4′) inhibited Pka 78-fold and activated Factor XI (FXIa) 350-fold less rapidly than AAT M358R. Eleven additional variants were expressed, restoring AAT residues stepwise. The most selective variant was AC (10–3/4′), which restored AAT residues from P2-P3′ compared to AC (10–4′), and inhibited Pka 1.9-fold more rapidly, and FXIa 1.6-fold less rapidly, for a gain in selectivity of 2.8-fold (p < 0.0001), without increasing the stoichiometry of inhibition (SI). The most active variant was AC (10−3), in which both the rate of Pka and FXIa inhibition were elevated relative to AAT M358R values, without SI elevation. Other variants exhibited slower reaction rates and/or elevated SI values. These results indicate that RCL exchanges can be productively employed to change serpin specificity and selectivity, but that the most effective exchanges may not be contiguous due to cooperativity between RCL residues.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 205-214"},"PeriodicalIF":4.1,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144170410","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}

引用次数: 0

pH-driven metabolic reprogramming in Bacillus velezensis 83 regulates metabolite synthesis and sporulation: A transcriptional approach for bioprocess development ph驱动的velezensis 83代谢重编程调节代谢物合成和孢子形成：生物过程发育的转录方法

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-05-26 DOI: 10.1016/j.jbiotec.2025.05.016

Lorena Yamileth Balón-Rosas, Agustín Luna-Bulbarela, Leobardo Serrano-Carreón, Enrique Galindo

{"title":"pH-driven metabolic reprogramming in Bacillus velezensis 83 regulates metabolite synthesis and sporulation: A transcriptional approach for bioprocess development","authors":"Lorena Yamileth Balón-Rosas, Agustín Luna-Bulbarela, Leobardo Serrano-Carreón, Enrique Galindo","doi":"10.1016/j.jbiotec.2025.05.016","DOIUrl":"10.1016/j.jbiotec.2025.05.016","url":null,"abstract":"<div><div><em>Bacillus velezensis</em> 83 (<em>Bv</em>83) spores are the active ingredient in Fungifree AB, an agricultural biocontrol agent whose industrial production is negatively affected by poly-γ-glutamic acid (γ-PGA) biopolymer synthesis at pH 6.8. At pH 5, γ-PGA production and sporulation are suppressed, but the latter is restored through pH-shift to 6.8 after glucose depletion. This work presents a comprehensive <em>Bv</em>83 spore bioprocess development through physiological and transcriptional analyses. At pH 5, <em>Bv</em>83 present a strategic response to pH stress by reducing its growth rate and redirecting energy towards survival rather than differentiation (downregulating lipopeptide and γ-PGA production genes) and maintaining SigB-mediated stress response rather than sporulation. Transcriptome analysis revealed downregulation of <em>comX</em> and <em>phrC</em> genes at pH 5, indicating sporulation limitation from insufficient signaling molecule production. Indeed, our results confirmed the essential role of the competence and sporulation factor (CSF) in differentiation of elongated cells into forespores. Furthermore, spent medium addition from high-cell-density cultures induced complete sporulation at pH 5, suggesting critical metabolite concentrations (besides CSF) are required. A pH-shift strategy during fed-batch cultivation suppressed γ-PGA synthesis, leading to enhanced mixing and oxygen transfer. Moreover, this strategy led to a 2.6-fold increase in spore productivity (7.86 ×10<sup>10</sup> spores L<sup>−1</sup> h<sup>−1</sup>) compared to a batch at pH 6.8, reducing operational costs. This research has identified pH-regulated metabolic networks, establishing a foundation for designing efficient, cost-effective industrial-scale <em>B. velezensis</em> fermentation strategies that comply with regulatory requirements for biocontrol applications.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 191-204"},"PeriodicalIF":4.1,"publicationDate":"2025-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144170592","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}

引用次数: 0

Control of alcoholic fermentation through modulation of nitrogen metabolism in Saccharomyces cerevisiae 通过调节酿酒酵母的氮代谢控制酒精发酵

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-05-20 DOI: 10.1016/j.jbiotec.2025.05.015

Naoki Akasaka , Yukiko Sugimoto , Takuma Kajihara , Hiroshi Takagi , Daisuke Watanabe

{"title":"Control of alcoholic fermentation through modulation of nitrogen metabolism in Saccharomyces cerevisiae","authors":"Naoki Akasaka , Yukiko Sugimoto , Takuma Kajihara , Hiroshi Takagi , Daisuke Watanabe","doi":"10.1016/j.jbiotec.2025.05.015","DOIUrl":"10.1016/j.jbiotec.2025.05.015","url":null,"abstract":"<div><div><em>Saccharomyces cerevisiae</em> sake strains exhibit high alcoholic fermentation performance. Comparative transcriptomic analysis revealed that the expression of genes required for nitrogen sensing and metabolism, including amino acid biosynthesis and uptake, was markedly lower in the sake strain than in the laboratory strain. Thus, we hypothesized that changes in nitrogen metabolism affect the fermentation capability of <em>S. cerevisiae</em>. To evaluate the impact of altered nitrogen metabolism on alcoholic fermentation, we focused on the transcription activators Gcn4p, Gln3p, and Gat1p, and the protein kinase Npr1p, all of which are key regulators controlling expression of genes for amino acid biosynthesis and uptake responding to nitrogen availability. Fermentation tests demonstrated that laboratory strain-derived single-deletion mutants of the regulator genes exhibited higher fermentation performance than the parental strain, which was accompanied by decrease in intracellular amino acid levels in the mutants. Disruption of the genes encoding glutamate dehydrogenases, which play a central role in nitrogen assimilation, also enhanced the fermentation rate. A Greatwall family protein kinase Rim15p inhibits alcoholic fermentation by diverting carbon flux from glycolysis to the synthesis of 1,3-β-glucan, a major cell wall component. Since the content of 1,3-β-glucan was unaffected by disruption of the regulator genes, the elevated fermentation performance of the disruptants was accomplished independently of the signaling pathway governed by Rim15p. The high fermentation rate of the disruptants might be attributed to increased carbon entry into glycolysis caused by the compromised biosynthesis of amino acids, which are synthesized from intermediary metabolites of glycolysis and tricarboxylic acid cycle.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 159-168"},"PeriodicalIF":4.1,"publicationDate":"2025-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144116338","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}

引用次数: 0

Investigating the influence of process parameters on the properties and refolding yield of single-chain variable fragment inclusion bodies 研究了工艺参数对单链可变片段包涵体性质和再折叠产率的影响。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-05-20 DOI: 10.1016/j.jbiotec.2025.05.014

Mohamed Elshazly , Benedikt Leeb , Eva Prada Brichtova , Florian Gisperg , Robert Klausser , Shilpa Vijayakumar , Bernhard Lendl , Martin Voigtmann , Matthias Berkemeyer , Oliver Spadiut , Julian Kopp

{"title":"Investigating the influence of process parameters on the properties and refolding yield of single-chain variable fragment inclusion bodies","authors":"Mohamed Elshazly , Benedikt Leeb , Eva Prada Brichtova , Florian Gisperg , Robert Klausser , Shilpa Vijayakumar , Bernhard Lendl , Martin Voigtmann , Matthias Berkemeyer , Oliver Spadiut , Julian Kopp","doi":"10.1016/j.jbiotec.2025.05.014","DOIUrl":"10.1016/j.jbiotec.2025.05.014","url":null,"abstract":"<div><div>Ever since the potential of inclusion bodies (IBs) has been recognized, substantial advances have been made towards understanding IB processes and enabling efficient and controlled development strategies. Still, the influence of the chosen upstream processing (USP) strategy on the properties of inclusion bodies (IBs) and their refolding performance remains poorly understood. This work aims to target this challenge by investigating the influence of two chosen USP parameters, namely the specific substrate uptake rate and the temperature during induction, on IB titer, IB properties, namely IB purity, size and secondary protein structure of the IBs, as well as refolding yield of single-chain variable fragment M (scFvM) IBs. Contrary to findings in the literature, USP conditions neither had a statistically significant effect on the aforementioned IB properties nor on the refolding yield, but could clearly alter the IB titer. Our results provide detailed analytical insights on the independence of IB properties from USP conditions for this protein, while increasing the volumetric IB productivity proved feasible through variations in USP parameters. Therefore, titer maximization appears to be the sole optimization strategy for scFvM IBs and these findings may also apply to other target proteins with similar structural properties.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 182-190"},"PeriodicalIF":4.1,"publicationDate":"2025-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144127738","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}

引用次数: 0

Identification and engineering of a sucrose synthase from Stevia rebaudiana for glycosylation applications 甜菊糖基化蔗糖合成酶的鉴定与工程研究。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-05-14 DOI: 10.1016/j.jbiotec.2025.05.011

Kai Chen , Yuan Liao , Xiaona Chen , Kecai Chen , Weicai Song , Liping Zhu , Yan Li , Honghua Jia

{"title":"Identification and engineering of a sucrose synthase from Stevia rebaudiana for glycosylation applications","authors":"Kai Chen , Yuan Liao , Xiaona Chen , Kecai Chen , Weicai Song , Liping Zhu , Yan Li , Honghua Jia","doi":"10.1016/j.jbiotec.2025.05.011","DOIUrl":"10.1016/j.jbiotec.2025.05.011","url":null,"abstract":"<div><div>Sucrose synthase (SuSy) is a unique glycosyltransferase that can be utilized in the production of nucleoside monosaccharides, such as diphosphate (UDP)-glucose, which serve as essential sugar donors for the glycosylation reactions catalyzed by UDP-dependent glycosyltransferases (UGTs). The selection of an appropriate SuSy coupled with a UGT is crucial for achieving the efficient synthesis of glycoside products. In this study, three candidate SuSy genes were identified from the transcriptome sequencing of <em>Stevia rebaudiana</em>, among which SrSUS1 was found to be expressed and active in <em>Escherichia coli</em>. The optimal temperature and pH for SrSUS1 were determined to be 55°C and pH 7.0, respectively. A variant SrSUS1<sub>T49A/L90P/V104E</sub> was generated based on a consensus sequence strategy, exhibiting a 3.6-fold increase in activity and the enhanced affinity for sucrose (<em>K</em><sub>m</sub> = 52.32 mM), as well as the improved thermal stability and catalytic efficiency. By coupling SrSUS1<sub>T49A/L90P/V104E</sub> with glycosyltransferase UGTAn85<sub>Q23E/N65D</sub> or UGT76G4, respectively, the production of 163.32 mM (43.63 g/L) of 2-phenylethyl-β-D-pyranoside and 72.29 mM (93.35 g/L) of rebaudioside M was achieved within 24 h in one-pot, two-enzyme fed-batch reactions. This study provides new insights into plant-derived SuSys and presents a promising biocatalyst for industrial glycosylation applications.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 169-181"},"PeriodicalIF":4.1,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144086101","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}

引用次数: 0

Production of astaxanthin with high purity and activity based on engineering improvement strategies 基于工程改进策略的高纯度、高活性虾青素的生产

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-05-14 DOI: 10.1016/j.jbiotec.2025.05.012

Chaogang Wang , Zeyu Hong , Mingjian Song , Hao Zheng , Qiaomian Zhou , Haihong Yang , Hui Li , Danqiong Huang

{"title":"Production of astaxanthin with high purity and activity based on engineering improvement strategies","authors":"Chaogang Wang , Zeyu Hong , Mingjian Song , Hao Zheng , Qiaomian Zhou , Haihong Yang , Hui Li , Danqiong Huang","doi":"10.1016/j.jbiotec.2025.05.012","DOIUrl":"10.1016/j.jbiotec.2025.05.012","url":null,"abstract":"<div><div>Here, astaxanthin production in <em>Escherichia coli</em> was systematically improved step by step. By introducing the additional copy of <em>CrtZ</em> and fusion complex of <em>CrtZ</em> and <em>CrtW</em>, astaxanthin content in cells increased from 0.10 mg/g to 0.16 mg/g and 0.63 mg/g DCW, respectively. Remolding the astaxanthin gene cluster by replacing the <em>PanCrtE</em> by <em>HpGGPPS3–1</em> and the fusion of <em>CrtZ</em> and <em>CrtW</em> increased astaxanthin content to 1.98 mg/g DCW. Further selecting the productive host and optimizing culture conditions dramatically increased astaxanthin content to 3.61 mg/g DCW. Subsequently, the fed-batch fermentation achieved the maximum yield of astaxanthin at 509.58 mg/L with the productivity of 7.72 mg/L/h and 5.91 mg/g DCW, covering 98.17 % of detected carotenoids. The chirality analysis assigned the same isomer of astaxanthin extracted from our fermentation system and <em>Haematococcus pluvialis</em>. Moreover, the radical and superoxide anion scavenging activity analysis revealed that astaxanthin achieved in this study performed better than natural astaxanthin extracted from <em>H. pluvialis</em> and chemical synthetic astaxanthin. This study provides a step-by-step example for bioengineering improvement of natural products in <em>E. coli</em> with high purity and activity.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 139-149"},"PeriodicalIF":4.1,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144067929","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}

引用次数: 0

Perspectives on the use of the CRISPR system in plants to improve recombinant therapeutic protein production 利用CRISPR系统在植物中提高重组治疗性蛋白生产的展望

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-05-13 DOI: 10.1016/j.jbiotec.2025.05.010

Edgar Trujillo, Carlos Angulo

{"title":"Perspectives on the use of the CRISPR system in plants to improve recombinant therapeutic protein production","authors":"Edgar Trujillo, Carlos Angulo","doi":"10.1016/j.jbiotec.2025.05.010","DOIUrl":"10.1016/j.jbiotec.2025.05.010","url":null,"abstract":"<div><div>The plant-based system is a promising platform for producing biotherapeutics due to its scalability, cost-effectiveness, and lower risk of contamination by human pathogens. However, several challenges remain, including optimizing yield, stability, functionality, and the immunogenic properties of recombinant proteins. In this context, this review explores the application of CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) technology to improve the production of recombinant therapeutic proteins in plants. Traditional tools and strategies for plant-based recombinant protein production are discussed, highlighting their limitations and the potential of CRISPR to overcome these boundaries. It delves into the components of the CRISPR-Cas system and its application in optimizing therapeutic protein function and yield. Major strategies include modifying glycosylation patterns to humanize plant-produced proteins, metabolic pathway engineering to increase protein accumulation, and the precise integration of transgenes into specific genomic loci to enhance expression stability and productivity. These advancements demonstrate how CRISPR system can overcome bottlenecks in plant molecular farming and enable the production of high-quality therapeutic proteins. Lastly, future trends and perspectives are examined, emphasizing ongoing innovations and challenges in the field. The review underscores the potential of CRISPR to reshape plant biotechnology and support the growing demand for recombinant therapeutics, offering new avenues for sustainable and efficient protein production systems.</div></div><div><h3>Key message</h3><div>CRISPR technology has the potential to improve plant-based therapeutic protein production by optimizing yield, stability, and humanization, overcoming bottlenecks, and enabling sustainable, efficient systems for recombinant biotherapeutics.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 111-123"},"PeriodicalIF":4.1,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144067928","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}

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