Biotechnology Journal最新文献_第10页

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-13

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IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-13

引用次数: 0

Integrated Fermentation Engineering Enables High-Level Leghemoglobin Production in Kluyveromyces marxianus via Metabolic Rewiring of Central Carbon Metabolism and Amino Acid Utilization 综合发酵工程通过中心碳代谢和氨基酸利用的代谢重新布线使克鲁维酵母高产出高水平的豆血红蛋白。

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-13 DOI: 10.1002/biot.70239

Hong Chen, Xinyu Jiang, Anxiang Chen, Junyuan Zen, Haiyan Ren, Yao Yu, Jungang Zhou, Jianye Xia, Hong Lu

{"title":"Integrated Fermentation Engineering Enables High-Level Leghemoglobin Production in Kluyveromyces marxianus via Metabolic Rewiring of Central Carbon Metabolism and Amino Acid Utilization","authors":"Hong Chen, Xinyu Jiang, Anxiang Chen, Junyuan Zen, Haiyan Ren, Yao Yu, Jungang Zhou, Jianye Xia, Hong Lu","doi":"10.1002/biot.70239","DOIUrl":"10.1002/biot.70239","url":null,"abstract":"<div>\u0000 \u0000 <p>Leghemoglobin (LBA), a hemoprotein critical for food industry applications, faces production challenges in microbial systems due to inefficient heme-globin assembly and metabolic constraints. Here, the food-safe yeast <i>Kluyveromyces marxianus</i> Tra-23 was validated as an excellent chassis, demonstrating robust growth and high LBA expression in bioreactor cultivation. Subsequently, key fermentation parameters—including medium composition, pH, temperature, and feeding strategy—were systematically optimized to enhance LBA biosynthesis. In 5-L bioreactors, the optimized processes elevated the LBA titer to 10.1 g/L, representing a 64.7% increase over initial conditions (6.13 g/L). Additionally, the specific glucose consumption rate (<i>q<sub>s</sub></i>) increased by approximately 10%, while the specific LBA production rate (<i>q<sub>p</sub></i>) rose over 100%. Notably, the optimized approach extended the cellular high productivity phase and enhanced the product yield from 0.011 g/g glucose to 0.026 g/g glucose. Furthermore, targeted metabolomics analysis of central carbon and amino acid metabolism provided mechanistic insights into the improved production performance. The optimized strategy reshaped the cellular metabolic network, enhanced carbon conversion efficiency toward LBA synthesis, and thereby improved the production performance of the microbial cell factory. These results highlight the potential of coordinated control of nutritional and physical parameters to enhance LBA biosynthesis in <i>K. marxianus</i>, providing a promising strategy for industrial production.</p>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 5","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147924734","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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

Engineering Yarrowia Lipolytica for De Novo Biosynthesis of Abscisic Acid 工程脂解耶氏菌对脱落酸的新生物合成。

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-13 DOI: 10.1002/biot.70232

Zhengxuan Shao, Qihang Chen, Wenbao Zhao, Qi Li, Xu Jia, Xiwen Li, Jingwen Zhou, Sha Xu

{"title":"Engineering Yarrowia Lipolytica for De Novo Biosynthesis of Abscisic Acid","authors":"Zhengxuan Shao, Qihang Chen, Wenbao Zhao, Qi Li, Xu Jia, Xiwen Li, Jingwen Zhou, Sha Xu","doi":"10.1002/biot.70232","DOIUrl":"10.1002/biot.70232","url":null,"abstract":"<div>\u0000 \u0000 <p>Abscisic acid (ABA) is a sesquiterpenoid plant hormone widely used in agriculture and the food industry. In <i>Yarrowia lipolytica</i>, ABA production relies on complex heterologous enzymatic pathways that have not been systematically engineered, which severely limits biosynthetic efficiency. Here, we establish <i>de novo</i> ABA biosynthesis in <i>Y. lipolytica</i>. We optimized carbon flux by up- and down-regulating key genes, and rationally engineered rate-limiting cytochrome P450 monooxygenases <i>Bc</i>ABA2 to improve pathway efficiency. Electron transfer was engineered by incorporating an NADPH regeneration module to enhance <i>Bc</i>ABA2 catalysis. Peroxisomal compartmentalization of the entire ABA pathway, followed by complementation of the defective tag to obtain a complete strain, increased ABA titer to 330.69 mg/L. The engineered strain was scaled up in a 5 L bioreactor, achieving an ABA titer of 2554.36 mg/L after 168 h, which is the highest titer reported to date for microbial fermentation. This work provides feasible enzyme and metabolic engineering strategies for microbial cell factory production of ABA and other terpenoids.</p>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 5","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147924612","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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

Engineering a Membrane-Anchored CreER Recombinase With Reduced Basal Activity for In Vitro Recombination 设计膜锚定CreER重组酶，降低基础活性用于体外重组。

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-12 DOI: 10.1002/biot.70236

Xiaotong Zeng, Xinyi Chen, Zishan Liang, Zhanze Yang, Junwei Li, Zhengrong Zhou, Chiju Wei, Aihua Mao

{"title":"Engineering a Membrane-Anchored CreER Recombinase With Reduced Basal Activity for In Vitro Recombination","authors":"Xiaotong Zeng, Xinyi Chen, Zishan Liang, Zhanze Yang, Junwei Li, Zhengrong Zhou, Chiju Wei, Aihua Mao","doi":"10.1002/biot.70236","DOIUrl":"10.1002/biot.70236","url":null,"abstract":"<div>\u0000 \u0000 <p>The CreER-loxP system is widely utilized for genetic cell lineage tracing and conditional gene function analysis. However, issues have arisen concerning the unintended labeling of nontarget cell populations, which complicate experimental data analysis and interpretation. In this study, we constructed an engineered CreER-loxP system in which a membrane‑localization motif was fused to the C‑terminus of Cre, thereby anchoring the fusion protein to the cell membrane and minimizing its spontaneous leakage into the nucleus. This membrane‑tethered CreER-loxP (mCreER-loxP) system substantially reduced nonspecific labeling in the absence of tamoxifen. Upon tamoxifen induction, the membrane‑localized Cre dissociated from the membrane and translocated into the nucleus, maintaining comparable recombination efficiency with the conventional CreER-loxP system. Notably, the membrane‑tethering design demonstrates enhanced performance under conditions of high expression levels and extended duration in vitro, even within the more stringent ERCreER-loxP labeling framework. Furthermore, mCreER exhibited reduced cytotoxicity relative to standard CreER. Collectively, these results indicate that the mCreER-loxP system provides improved temporal and spatial precision for applications in cell lineage tracing and genetic engineering.</p>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 5","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147872495","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-12

引用次数: 0

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-12 DOI: 10.1002/biot.70235

Dongliang Chen, Qiongqiong Zhang, Zhe Lang, Zheyu Wang, Xiang Zheng, Mingyue Fang, Hang Zhou

{"title":"Dual-Similarity Driven Just-in-Time Modeling Enables Precise Real-Time Monitoring of Cell Cultures via Dielectric Spectroscopy","authors":"Dongliang Chen, Qiongqiong Zhang, Zhe Lang, Zheyu Wang, Xiang Zheng, Mingyue Fang, Hang Zhou","doi":"10.1002/biot.70235","DOIUrl":"10.1002/biot.70235","url":null,"abstract":"<div>\u0000 \u0000 <p>The biopharmaceutical industry is increasingly focused on shortening development timelines to accelerate progression into clinical stages, thereby improving patient outcomes and boosting market competitiveness. Dielectric spectroscopy, a promising process analytical technology (PAT), offers enhanced monitoring, and control of mammalian cell culture bioprocesses, but its practical application is constrained by the limited accuracy of existing models due to sparse historical data within compressed development periods. Just-in-time modeling (JITM) has recently emerged as a soft sensing approach balancing model accuracy and generalizability. However, its application in dielectric spectroscopy remains unreported, mainly due to the absence of suitable similarity metrics. This study firstly proposes a novel dual-similarity driven JITM framework tailored for dielectric spectroscopy-based modeling. Various similarity strategies were evaluated through validation study, then the generalizability of JITM was demonstrated using multiple local model algorithms, while its effectiveness was further confirmed using a testing application involving real-time predictions of VCD. Compared to traditional techniques, the proposed JITM framework exhibited reliable and accurate VCD monitoring across diverse cell lines and culture conditions, showing great potential for industrial applications, especially in agile biomanufacturing where historical data and development time are limited.</p>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 5","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147872498","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-12

引用次数: 0

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-07

引用次数: 0

A New Conditional Transcription Factor for Use in Toxoplasma Gondii 一种新的用于刚地弓形虫的条件转录因子。

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-07 DOI: 10.1002/biot.70240

Mohammad Farouq Sharifpour, Shadi Khadiv, Alessandro D. Uboldi, Amalie A. Jayawickrama, Sarah Manning, Jonathan Whittall, Cynthia D. K. Bottema, John T. Ellis, Daniel K. Howe, Danny W. Wilson, Farhid Hemmatzadeh, Christopher J. Tonkin, Milton M. McAllister

{"title":"A New Conditional Transcription Factor for Use in Toxoplasma Gondii","authors":"Mohammad Farouq Sharifpour, Shadi Khadiv, Alessandro D. Uboldi, Amalie A. Jayawickrama, Sarah Manning, Jonathan Whittall, Cynthia D. K. Bottema, John T. Ellis, Daniel K. Howe, Danny W. Wilson, Farhid Hemmatzadeh, Christopher J. Tonkin, Milton M. McAllister","doi":"10.1002/biot.70240","DOIUrl":"10.1002/biot.70240","url":null,"abstract":"<p><i>Toxoplasma gondii</i> is an intracellular eukaryotic parasite in the phylum Apicomplexa. Reversible, conditional control of gene transcription was achieved previously in this organism using a tet-off system that was controlled by tetracycline. Although revolutionary for the field at the time, that system in <i>Toxoplasma</i> yielded modest signal-to-noise ratios (SNR) and proved to be leaky in the “off” state (i.e., it suffered from high background expression). Here we report the development of a new reversible and robust Conditional Transcription Factor (CTF) that is controlled by rapamycin. Although we originally intended to design a dimerizable drug-on transcription control system, the CTF unexpectedly functions in a drug-off fashion. In stably-transfected tachyzoites, CTF regulates a fluorescent reporter gene in <i>Toxoplasma</i> to achieve almost no background expression in the presence of rapamycin (i.e. near-zero leakiness), while in the absence of rapamycin 84% of tachyzoites express EYFP in comparison with 87% of constitutively-expressed positive controls, with 59% mean fluorescence intensity relative to positive controls. Conditional regulation of the median fluorescence intensity of the reporter gene reached an impressive SNR averaging 1489, approximating an “on/off” switch. The modular design of the CTF is expected to facilitate application to varied genes, incremental modifications, and adaptation for use in other apicomplexan organisms.</p>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 5","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13150947/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147831371","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0