Biotechnology Journal最新文献_第3页

Issue Information: Biotechnology Journal 9/2026 期刊信息：Biotechnology Journal 9/2026

IF 3.7 3区生物学

Biotechnology Journal Pub Date : 2026-08-27 DOI: 10.1002/biot.70293

引用次数: 0

Expanding Transposase Technology to Regulatory Noncoding RNAs for Stable Expression in CHO Cells 扩展转座酶技术调控非编码rna在CHO细胞中的稳定表达。

IF 3.7 3区生物学

Biotechnology Journal Pub Date : 2026-08-23 DOI: 10.1002/biot.70299

Sonja Lochmueller, Linus Weiss, Kerstin Otte

{"title":"Expanding Transposase Technology to Regulatory Noncoding RNAs for Stable Expression in CHO Cells","authors":"Sonja Lochmueller, Linus Weiss, Kerstin Otte","doi":"10.1002/biot.70299","DOIUrl":"10.1002/biot.70299","url":null,"abstract":"Stable expression of regulatory non-coding RNAs enables targeted cell engineering in Chinese hamster ovary (CHO) cells but commonly relies on random integration, resulting in variable expression. Here, we establish a transposase-mediated platform for stable microRNA (miRNA) expression in CHO cells and demonstrate the first application of a DNA transposon system for long-term miRNA genome integration. Using miR-3096b-5p, we compared piggyBac-mediated integration with conventional random integration in antibody-producing CHO cells. Transposase-mediated integration enabled rapid generation of stable cell pools with higher fractions of GFP-expressing cells. Both integration strategies supported comparable growth, viability, and antibody production in batch cultures, indicating that transposase-mediated miRNA integration does not compromise bioprocess performance. At the molecular level, transposase-mediated integration resulted in significantly higher transgene copy numbers and markedly increased miRNA expression, translating into stronger and more consistent regulation of the target genes Fuk and Gmds. By combining high integration efficiency and expression stability with the regulatory potential of miRNAs, this approach expands the applicability of transposase systems beyond protein-coding transgenes and provides a robust platform for precise and durable post-transcriptional gene regulation in CHO cell engineering.","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 8","pages":""},"PeriodicalIF":3.7,"publicationDate":"2026-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13501212/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148808062","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

Evolutionary Adaptation and Targeted Metabolic Engineering Synergistically Improve Methanol-Derived Single-Cell Protein Production in Pichia pastoris 进化适应和靶向代谢工程协同提高毕赤酵母甲醇来源的单细胞蛋白产量。

IF 3.7 3区生物学

Biotechnology Journal Pub Date : 2026-08-23 DOI: 10.1002/biot.70300

Chong Xie, Chengchao Zhu, Junze Liu, Huiqi Sun, Jingliang Xu, Qi Liu, Yajie Wang

{"title":"Evolutionary Adaptation and Targeted Metabolic Engineering Synergistically Improve Methanol-Derived Single-Cell Protein Production in Pichia pastoris","authors":"Chong Xie, Chengchao Zhu, Junze Liu, Huiqi Sun, Jingliang Xu, Qi Liu, Yajie Wang","doi":"10.1002/biot.70300","DOIUrl":"10.1002/biot.70300","url":null,"abstract":"<div>\u0000 \u0000 Methanol is a promising renewable C1 feedstock for sustainable single-cell protein (SCP) production. However, its inherent cytotoxicity and metabolic trade-offs between cell growth and protein synthesis remain significant bottlenecks. Here, we established an “evolutionary-rational” dual-driven paradigm to construct a high-yield Pichia pastoris chassis. Through UV mutagenesis and adaptive laboratory evolution, we developed a highly tolerant strain A40, capable of growing in 70 g/L methanol. Notably, at 30 g/L methanol, A40 achieved a 3.4-fold higher maximum biomass than the wild-type. Whole-genome resequencing and reverse genetics revealed that this superior performance stems from a multi-gene synergistic network rather than a single dominant mutation. To further optimize SCP production, we rationally co-overexpressed nitrogen assimilation genes (GLN1, GDH1) and a translation elongation factor (PpeEF3) in the A40 background. This targeted metabolic engineering effectively redirected carbon flux toward protein biosynthesis. The engineered strain A40-2Ge3 achieved a peak intracellular crude protein content of 67.9% and a 51.3 g/L total titer in a 5-L bioreactor, representing a 23.1% increase over the wild-type strain. Collectively, this study provides deep insights into the synergistic mechanisms of methanol adaptation and establishes an efficient, scalable strategy for sustainable SCP production from C1 feedstocks.\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 8","pages":""},"PeriodicalIF":3.7,"publicationDate":"2026-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148808047","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

Pressure Dependent Changes in CO2-Concentrations Modulate the 3-Hydroxyvalerate Fraction in Terpolymeric Polyhydroxyalkanoates 压力相关的co2浓度变化调节三聚体聚羟基烷酸酯中3-羟戊酸酯的组分

IF 3.7 3区生物学

Biotechnology Journal Pub Date : 2026-08-20 DOI: 10.1002/biot.70298

Birk Achenbach, Pravesh Tamang, Günter E. M. Tovar, Susanne Zibek

{"title":"Pressure Dependent Changes in CO2-Concentrations Modulate the 3-Hydroxyvalerate Fraction in Terpolymeric Polyhydroxyalkanoates","authors":"Birk Achenbach, Pravesh Tamang, Günter E. M. Tovar, Susanne Zibek","doi":"10.1002/biot.70298","DOIUrl":"https://doi.org/10.1002/biot.70298","url":null,"abstract":"Polyhydroxyalkanoates (PHAs), especially their copolymeric forms, such as poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-4-hydroxyvalerate) (PHBVV), have promising properties for replacing petrochemical-derived plastics. However, the research focuses mostly on producing PHA from a specific substrate or waste-stream, while the reproducibility of a precise polymer composition is seldom targeted. This study systematically focuses on the influence of pressure conditions on the production of terpolymeric PHBVV by Cupriavidus necator NCIMB 11599 using levulinic acid (LA). Three different process setups were evaluated to assess the impact of hydrostatic pressure (ΔpHydro) and applied pressure (ΔpVessel) compared to the process at ambient pressure. Differences were detected regarding carbon utilization (YX/Gluc & YPHA/LA) and polymer composition. The YX/Gluc increased in the pilot scale compared to the technical scale from 0.31 ± 0.01 gX gGluc−1 to 0.41 ± 0.01 gX gGluc−1. Conversely, as ΔpVessel increased, the YPHA/LA decreased from 0.35 ± 0.02 gPHA gLA−1 to 0.23 ± 0.02 gPHA gLA−1, while the corresponding 3HV contents increased from 48 ± 1%(w/w) to 69 ± 2%(w/w). These changes were consistent with pressure-dependent variations in CO2 concentration and a potentially increased carbon fixation via the energy-intensive CBB cycle. This indicates that the polymer composition and cell vitality can be adjusted via the CO2 concentration, a factor that should be taken into account when further scaling up of PHA production processes.","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 8","pages":""},"PeriodicalIF":3.7,"publicationDate":"2026-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/biot.70298","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148784733","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

Endochitinase Overexpression in Transgenic Tomato Plants Improves Resistance to Phytopathogens 内几丁质酶在转基因番茄植株中的过表达提高了对植物病原体的抗性

IF 3.7 3区生物学

Biotechnology Journal Pub Date : 2026-08-20 DOI: 10.1002/biot.70297

Lucas José de Sousa, Ana Carolina Mendes Bezerra, Ivonaldo Reis Santos, Ana Clara Muniz Martins Vilela, Osmundo Brilhante de Oliveira Neto, Glaucia Barbosa Cabral, Alice Maria Quezado-Duval, Pedro Ricardo Vieira Hamann, Eliane Ferreira Noronha, Luiz Eduardo Bassay Blum, Angela Mehta

{"title":"Endochitinase Overexpression in Transgenic Tomato Plants Improves Resistance to Phytopathogens","authors":"Lucas José de Sousa, Ana Carolina Mendes Bezerra, Ivonaldo Reis Santos, Ana Clara Muniz Martins Vilela, Osmundo Brilhante de Oliveira Neto, Glaucia Barbosa Cabral, Alice Maria Quezado-Duval, Pedro Ricardo Vieira Hamann, Eliane Ferreira Noronha, Luiz Eduardo Bassay Blum, Angela Mehta","doi":"10.1002/biot.70297","DOIUrl":"https://doi.org/10.1002/biot.70297","url":null,"abstract":"Engineering broad-spectrum disease resistance remains a major challenge in crop improvement because most resistance genes are effective against a limited range of pathogens. Pathogenesis-related (PR) proteins, particularly endochitinases, are promising targets for enhancing plant immunity due to their antimicrobial activity and ability to activate defense responses. Here, we generated transgenic tomato (Solanum lycopersicum L.) plants overexpressing an endochitinase gene to assess their potential for enhanced resistance against bacterial and fungal pathogens. Three independent overexpression lines and the wild type were evaluated for transgene expression, chitinase activity, defense-related gene expression, and resistance after inoculation with Xanthomonas euvesicatoria pv. perforans (Xep) and Sclerotinia sclerotiorum. The transgene was strongly expressed in all transgenic lines, whereas chitinase activity increased by up to fivefold compared with the wild type. Endochitinase overexpression activated defense responses, with marked induction of PR1 expression, reaching a maximum increase of over 480-fold, and enhanced expression of defense-associated genes, including SlACS and SlACO. Compared with the wild type, the most responsive transgenic lines showed reductions of up to 79.7% in bacterial spot severity and 85.6% in white mold severity. These findings demonstrate that endochitinase overexpression activates defense signaling and improves broad-spectrum resistance, representing a promising strategy for developing disease-resistant tomato cultivars.","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 8","pages":""},"PeriodicalIF":3.7,"publicationDate":"2026-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/biot.70297","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148784732","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

Anion-Exchange Membrane Chromatography for the Separation of Empty and Full Adeno-Associated Viral Capsids 阴离子交换膜色谱法分离空和满腺相关病毒衣壳。

IF 3.7 3区生物学

Biotechnology Journal Pub Date : 2026-08-17 DOI: 10.1002/biot.70295

Luca Ossi, Samuele Delfino, Helena Marie, Michael Schulte, Mattia Sponchioni

{"title":"Anion-Exchange Membrane Chromatography for the Separation of Empty and Full Adeno-Associated Viral Capsids","authors":"Luca Ossi, Samuele Delfino, Helena Marie, Michael Schulte, Mattia Sponchioni","doi":"10.1002/biot.70295","DOIUrl":"10.1002/biot.70295","url":null,"abstract":"The separation of empty and full adeno-associated viruses (AAVs) is particularly critical because of the similarity between the two. Anion-exchange chromatography represents a scalable tool, leveraging their slight difference in isoelectric point. However, traditional resin chromatography can only deliver poor resolution, mainly due to the restricted accessibility of the capisds to the bead pores. To cover this gap, in this work, we designed and optimized the separation of empty and full AAVs using anion-exchange membrane chromatography. The role played by flowrate, gradient slope and wash step parameters on the chromatographic resolution was first investigated with two consecutive designs of experiments (DoE). The response surfaces indicated that high process flowrates, that is 15 membrane volumes (MVs)/min, with a gradient from 115 mM to 250 mM sodium acetate in 350 MVs maximize the chromatographic resolution. Then, preparative experiments showed that fast loading is possible, with residence time as short as 24 s, without product breakthrough. Moreover, membrane loading up to 1.2E17 viral particles (vp)/Lmembrane could be reached, providing one order of magnitude higher productivity with respect to standard column operations. With these results, we demonstrated the great potential of anion-exchange membrane chromatography in the polishing of AAVs, overcoming traditional limitations of packed-bed operations.","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 8","pages":""},"PeriodicalIF":3.7,"publicationDate":"2026-08-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/biot.70295","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148759550","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

Impact of Cistron Topology and Bicistronic Vector Design in the piggyBac Transposon System for Improved Antibody Expression 顺反子拓扑结构和双顺反子载体设计对piggyBac转座子系统改善抗体表达的影响。

IF 3.7 3区生物学

Biotechnology Journal Pub Date : 2026-08-16 DOI: 10.1002/biot.70296

Jason Vitko, Thomas King, Efecan Aral, Steven McLellan, Hope Divello, Sara Maimouni, Tiffany McLamarrah, Jennifer Tedstone, Victor Cairns, Christine DeMaria, John J. Scarcelli

{"title":"Impact of Cistron Topology and Bicistronic Vector Design in the piggyBac Transposon System for Improved Antibody Expression","authors":"Jason Vitko, Thomas King, Efecan Aral, Steven McLellan, Hope Divello, Sara Maimouni, Tiffany McLamarrah, Jennifer Tedstone, Victor Cairns, Christine DeMaria, John J. Scarcelli","doi":"10.1002/biot.70296","DOIUrl":"10.1002/biot.70296","url":null,"abstract":"The use of transposon-based technologies in cell line development has gained significant traction over the past decade. This study investigates the piggyBac transposon system combined with bicistronic vectors for monoclonal antibody expression, helping address the complex demands of multi-specific antibody formats that require multiple cistrons. We systematically evaluated bicistronic vector modifications within the piggyBac framework, including promoter sequences, reporter placements, and cassette configurations, and compared performance to piggyBac/single-gene vector co-transfection methods. Our findings demonstrate that double human CMV promoter configurations driving both heavy and light chain genes significantly enhanced pool productivity (2 to 2.5-fold) and reporter expression compared to separate promoter designs. Among tested cassette arrangements, the Light chain-GS-Heavy chain configuration yielded optimal productivity (1.5 to 6-fold) and superior heavy chain/light chain RNA transcript ratios (1.4 to 3.8-fold and 1.3 to 6.1-fold, respectively). While unfed batch conditions showed comparable productivities between optimized piggyBac/bicistronic and piggyBac/single-gene systems, the bicistronic approach exhibited superior productivity performance under fed-batch conditions (1.5-fold), with the resulting clones demonstrating significantly high productivity (top clone 9.6 g/L). This represents the first reported use of LC-selection marker-HC topology for monoclonal antibody expression, establishing a foundation for improved therapeutic biologic production through multicistronic vector systems combined with the piggyBac transposon strategy.","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 8","pages":""},"PeriodicalIF":3.7,"publicationDate":"2026-08-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/biot.70296","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148757038","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

Immunological Evaluation of mRNA-LNP Vaccines Targeting Conserved S2 and NSP3 Domains of SARS-CoV-2 靶向SARS-CoV-2保守S2和NSP3结构域的mRNA-LNP疫苗的免疫学评价

IF 3.7 3区生物学

Biotechnology Journal Pub Date : 2026-08-16 DOI: 10.1002/biot.70294

Feroza Begum, Md Hasan Mallik, Subhadip Ghora, Upasana Ray

{"title":"Immunological Evaluation of mRNA-LNP Vaccines Targeting Conserved S2 and NSP3 Domains of SARS-CoV-2","authors":"Feroza Begum, Md Hasan Mallik, Subhadip Ghora, Upasana Ray","doi":"10.1002/biot.70294","DOIUrl":"10.1002/biot.70294","url":null,"abstract":"<div>\u0000 \u0000 The evolution of SARS-CoV-2 necessitates next-generation vaccine strategies targeting conserved viral elements to ensure broad and durable protection. In this study, we designed and evaluated novel mRNA-based vaccines encoding conserved domains of SARS-CoV-2, including the spike S2 subunit, NSP3 Macrodomain I (Mac I), and papain-like protease (PLpro). These mRNAs were synthesized, cloned, and encapsulated into lipid nanoparticles (LNPs) with 70%–80% efficiency and favorable physicochemical properties. In vitro characterization showed efficient antigen expression with 60%–80% cell survival. Immunization of BALB/c mice induced strong humoral responses, particularly against S2 and Mac I, marked by IgG titers, class switching, and durable antibody presence up to 84 days post-booster. PLpro elicited minimal antibody responses, likely due to limited stability or intrinsic low immunogenicity. Combinatorial LNPs encoding multiple antigens maintained robust expression and elicited comparable immune responses despite dose splitting. Anti-S2 antibodies effectively inhibited spike protein uptake in vitro. Importantly, the selected antigenic regions exhibited high sequence conservation across major SARS-CoV-2 variants, underscoring their broad-spectrum vaccine potential. These findings support further development of domain-targeted, multi-antigen mRNA vaccines to enhance cross-variant protection and T-cell-mediated immunity.\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 8","pages":""},"PeriodicalIF":3.7,"publicationDate":"2026-08-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148759573","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

Spatial Engineering of Yarrowia lipolytica for the Biosynthesis of Capsanthin 聚脂耶氏菌生物合成辣椒素的空间工程研究。

IF 3.7 3区生物学

Biotechnology Journal Pub Date : 2026-08-16 DOI: 10.1002/biot.70291

Jialei Zhang, Ling Sun, Liang Zhang, Chongyang Din, Zhenghua Gu, Lei Chen, Sha Xu

{"title":"Spatial Engineering of Yarrowia lipolytica for the Biosynthesis of Capsanthin","authors":"Jialei Zhang, Ling Sun, Liang Zhang, Chongyang Din, Zhenghua Gu, Lei Chen, Sha Xu","doi":"10.1002/biot.70291","DOIUrl":"10.1002/biot.70291","url":null,"abstract":"<div>\u0000 \u0000 Capsanthin is a high-value tetraterpenoid widely utilized in the food, cosmetic, and pharmaceutical industries. However, its traditional production via plant extraction and chemical synthesis faces severe sustainability and cost constraints. The oleaginous yeast Yarrowia lipolytica (Y. lipolytica) offers a superior chassis due to its robust mevalonate pathway and abundant lipid droplets for pigment sequestration. In this study, we systematically engineered Y. lipolytica for the de novo biosynthesis of capsanthin. By heterologously co-expressing of Capsicum annuum capsanthin/capsorubin synthase (CaCCS) and Arabidopsis thaliana zeaxanthin epoxidase (AtZEP), we achieved the first reported biosynthesis of capsanthin in Y. lipolytica. Combinatorial screening revealed that full-length sequences of CaCCS and AtZEP yielded optimal production of 6.92 mg/L. To further enhance the productivity of capsanthin, we evaluated spatial compartmentalization strategies. The results demonstrated that targeting the biosynthetic enzymes to the peroxisome via Ser–Lys–Leu (SKL) tags successfully increased the titer to 8.8 mg/L by leveraging its specialized lipophilic and redox microenvironment. Finally, a 5-L fed-batch fermentation was conducted, achieving a maximum capsanthin titer of 48.15 mg/L. This work provides a sustainable and scalable alternative to plant extraction and chemical synthesis for capsanthin production.\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 8","pages":""},"PeriodicalIF":3.7,"publicationDate":"2026-08-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148757176","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

Metabolic Engineering of Komagataella Phaffii for de Novo Synthesis of Retinol From Methanol 法菲Komagataella Phaffii在甲醇从头合成视黄醇中的代谢工程。

IF 3.7 3区生物学

Biotechnology Journal Pub Date : 2026-08-16 DOI: 10.1002/biot.70288

Yanxuan Wu, Chengan Liu, Shuli Liang, Yuchen Jiang, Xinying Zhang, Yuqi Zhou, Ying Lin

{"title":"Metabolic Engineering of Komagataella Phaffii for de Novo Synthesis of Retinol From Methanol","authors":"Yanxuan Wu, Chengan Liu, Shuli Liang, Yuchen Jiang, Xinying Zhang, Yuqi Zhou, Ying Lin","doi":"10.1002/biot.70288","DOIUrl":"10.1002/biot.70288","url":null,"abstract":"<div>\u0000 \u0000 Retinol, a derivative of vitamin A with potent antioxidant and therapeutic properties, is in high market demand. In response to the low productivity of conventional methods, metabolic engineering has been explored for microbial retinol production. However, systematic engineering strategies for high-level retinol synthesis in Komagataella phaffii remain limited. In this study, the methylotrophic yeast K. phaffii was developed as an engineered chassis for efficient de novo retinol biosynthesis. Based on a previously constructed β-carotene-producing strain, β-carotene-15,15′-dioxygenase (Blh) and retinol dehydrogenase (RDH12) were screened and introduced to establish the synthetic pathway of retinol. To increase precursor supply, key genes in the β-carotene biosynthetic pathway were overexpressed. The mevalonate (MVA) pathway was further optimized, and central carbon metabolism was reprogrammed to enhance metabolic flux toward retinol. Transport engineering was also performed to improve retinol secretion. Several candidate transporters were overexpressed, and the protein encoded by chr1-4_0619 in K. phaffii was identified as an endogenous retinol transporter. The final engineered strain produced 3.38 g/L retinol with BHT supplementation in fed-batch fermentation using a 1.5 L bioreactor. This work represents de novo microbial synthesis of retinol from a one-carbon feedstock, demonstrating the formidable potential of K. phaffii as a sustainable chassis for retinol production.\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 8","pages":""},"PeriodicalIF":3.7,"publicationDate":"2026-08-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148759051","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