Journal of biotechnology最新文献

{"title":"Performance of a pilot scale pressurized deep-jet gas bioreactor for SCP production with Cupriavidus necator H16","authors":"Kay Domenico Novak ,&nbsp;Petra Heidinger ,&nbsp;Daniel Schwendenwein ,&nbsp;Florian Haneder ,&nbsp;Daniel Martinjak ,&nbsp;Christoph Hochenauer ,&nbsp;Helmut Schwab ,&nbsp;Christoph Reisinger","doi":"10.1016/j.jbiotec.2025.08.013","DOIUrl":"10.1016/j.jbiotec.2025.08.013","url":null,"abstract":"<div><div>Single cell protein (SCP) production has the potential for significant contribution to global protein supply. The ground-breaking approach presented here demonstrates the industrial potential of <em>Cupriavidus necator</em> H16, a chemolithoautotrophic bacterium uniquely capable of converting CO₂ into high quality SCP. The study, conducted in a custom 300-L pressurized deep-jet bioreactor optimized for extremely efficient gas-liquid transfer, provides crucial insights for scalable SCP production. It highlights the significant influence of the inoculum type on the efficiency of fermentation. Heterotrophic inoculum caused a 10-hour lag phase, which limited the efficiency of the fermentation process, while chemolithoautotrophic inoculum achieved almost immediate productivity without a lag phase. This marked difference in performance underscores the importance of inoculum selection in maximizing yield. Fermentation with chemolithoautotrophic inoculum reached 300 g L⁻¹ cell wet weight (CWW) in only 35 h – a remarkable 44 % faster than the 63 h required with heterotrophic inoculum. By repeated removal of larger broth samples, the biomass concentration could be further increased to 360 g L⁻¹ CWW, which corresponds to to 90 g L⁻¹ cell dry weight (CDW). This demonstrates the feasibility of continuous operation and scalable production. The highest gas transfer and consumption rates were observed approximately 35 h after the start of fermentation: H₂ at 0.82 kg h⁻¹, O₂ at 4.1 kg h⁻¹and CO₂ at 1.95 kg h⁻¹. These high transfer rates underline the efficiency of the bioreactor in gas utilization, a critical factor for industrial-scale applications. The biomass showed a very high crude protein content of 80 % with an amino acid profile comparable to that of commercial fishmeal. This positions the SCP as a high-quality, sustainable alternative for application in aquaculture and poultry feed, meeting global protein needs while reducing environmental impact.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"408 ","pages":"Pages 1-14"},"PeriodicalIF":3.9,"publicationDate":"2025-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144933787","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"3-hydroxypropionic acid production from Brewer’s spent grain with an engineered Issatchenkia orientalis","authors":"Linjing Jia ,&nbsp;Deokyeol Jeong ,&nbsp;Mairui Zhang ,&nbsp;Eun Joong Oh ,&nbsp;Jikai Zhao","doi":"10.1016/j.jbiotec.2025.08.009","DOIUrl":"10.1016/j.jbiotec.2025.08.009","url":null,"abstract":"<div><div>3-Hydroxypropionic acid (3-HP) is a versatile platform chemical with wide-ranging industrial applications. This study presents a proof-of-concept approach for producing 3-HP from brewer’s spent grain (BSG) using a novel one-pot CaO pretreatment method and an engineered, acid-tolerant <em>Issatchenkia orientalis</em> IoDY01H strain. The effects of acid type for pH adjusting of pretreated slurry, nitrogen supplementation, NaHCO₃ addition, and BSG deproteinization were evaluated. Results showed that using pure glucose and xylose as carbon sources, yeast extract-peptone (YPDX) medium enhanced 3-HP titers by 13.3 % compared to synthetic complete (SCDX) medium. NaHCO₃ supplementation further boosted the 3-HP titer to 7.8 g/L, representing a 7.8 % increase over YPDX alone. In BSG hydrolysates, pH adjustment with H₂SO₄ resulted in higher 3-HP production (7.4 g/L) compared to H₃PO₄ (6.8 g/L). Moreover, supplementation with NaHCO₃ further increased the 3-HP titer to 7.7 g/L, while yeast extract-peptone reduced the 3-HP titer to 6.7 g/L by diverting flux toward strain growth and ethanol formation. Notably, deproteinization of BSG improved xylose recovery and achieved the highest 3-HP titer of 8.7 g/L without nitrogen supplementation. This work provides insights into integrating one-pot pretreatment and hydrolysis and robust fermentation without external nitrogen to produce value-added products from biomass feedstocks.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"407 ","pages":"Pages 70-77"},"PeriodicalIF":3.9,"publicationDate":"2025-08-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144903019","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The CRISPR Cas patent files, part 4: All back to zero, think again!","authors":"Ulrich Storz","doi":"10.1016/j.jbiotec.2025.08.001","DOIUrl":"10.1016/j.jbiotec.2025.08.001","url":null,"abstract":"<div><div>The epic patent disputes regarding CRISPR Cas9 have been keeping the Biotech community abuzz over the last 10 years. The unclear situation has created, and keeps creating, serious uncertainties among users of this groundbreaking technology. This article gives an overview of the dispute’s history, and explains the actual state of the debate.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"407 ","pages":"Pages 78-86"},"PeriodicalIF":3.9,"publicationDate":"2025-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144913262","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization of Single Cell Protein produced by Cupriavidus necator grown on various nitrogen and carbon sources.","authors":"Siwar Ismail, Géraldine Giacinti, Christine Delagado Raynaud, Sandrine Alfenore, Stéphane E Guillouet, Nathalie Gorret","doi":"10.1016/j.jbiotec.2025.08.007","DOIUrl":"https://doi.org/10.1016/j.jbiotec.2025.08.007","url":null,"abstract":"<p><p>Microorganisms can serve as Single Cell Proteins (SCP) used as an alternative and non-conventional protein source to meet the protein shortage and increasing demand. The objective of this research was to characterize growth and biomass composition of the bacterium Cupriavidus necator, cultivated on various nitrogen (ammonium sulfate and urea) and carbon (glucose, formic acid and oleic acid) sources. Under well-controlled conditions, a mono-factorial approach was employed to study the impact of the nature of the substrates on biomass growth and composition in bioreactors. Cultures were performed in batch culture mode, except for formic acid for which a fed-batch mode was selected due to its toxicity on cell growth. Results were compared in terms of growth, cell morphology, cell membrane permeability, biomass composition in proteins, nucleic acids and polyhydroxyalkanoates. The results showed high protein amounts and an impact of substrate on biomass composition. Independently, ammonium sulfate, as nitrogen source, and formic acid, as carbon source provided better results in term of growth and biomass composition. The highest total proteins (83%_DW), the lowest PHA amount (1.7%_DW) and the lowest changes in cell size and granularity profiles were obtained using the organic acid, showing the more attractive SCP composition. Despite interesting amino acid contents, the results showed that their distribution is affected by the nature of the substrates.</p>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":" ","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144859238","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Hydrolysis of chlorogenic acid by a novel GDSL lipase/esterase from sunflower","authors":"Cui Guo,&nbsp;Vanna Nguyen,&nbsp;Ashley Tseng,&nbsp;Yuheng Lin","doi":"10.1016/j.jbiotec.2025.08.006","DOIUrl":"10.1016/j.jbiotec.2025.08.006","url":null,"abstract":"<div><div>GDSL esterases/lipases are a diverse group of hydrolytic enzymes with broad substrate specificity. However, none of them have been reported to catalyze the hydrolysis of chlorogenic acid (CGA), a key plant metabolite. To date, all known CGA-hydrolyzing enzymes have been identified in microorganisms and belong to the classical esterase family. In this study, we report the identification and characterization of a novel GDSL-type enzyme HaCAE from <em>Helianthus annuus</em> (sunflower), which exhibits robust CGA-hydrolyzing activity. HaCAE was successfully expressed and secreted in recombinant <em>Pichia pastoris</em>. The kinetic parameters of HaCAE were determined through an <em>in vitro</em> enzyme assay. Whole-cell biocatalysis demonstrated remarkable enzymatic efficiency, achieving hydrolysis of 17.31 mM of CGA within 24 h. <em>In silico</em> analysis provided insights into the enzyme’s catalytic mechanism and substrate selectivity. To the best of our knowledge, HaCAE is the first plant-derived CGA-hydrolyzing enzyme. Its high catalytic activity underscores its potential for food processing applications aimed at reducing CGA content and offers valuable biochemical insights into CGA metabolism in plants.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"407 ","pages":"Pages 43-47"},"PeriodicalIF":3.9,"publicationDate":"2025-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144816733","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhancing the extraction of C-phycocyanin and water-soluble metabolites from Arthrospira platensis using pulsed electric field technology","authors":"Njomza Ibrahimi ,&nbsp;Ángela Sánchez-Quintero ,&nbsp;Anna Unterholzner ,&nbsp;Aurélien Parsy ,&nbsp;Amandine Adrien ,&nbsp;Yves Le Guer ,&nbsp;Antoine Silvestre de Ferron ,&nbsp;Susana C.M. Fernandes ,&nbsp;Laurent Pécastaing ,&nbsp;Jean-Baptiste Beigbeder","doi":"10.1016/j.jbiotec.2025.08.005","DOIUrl":"10.1016/j.jbiotec.2025.08.005","url":null,"abstract":"<div><div>Pulsed Electric Fields (PEF) technique has emerged as a promising approach to extract molecules of interest from different biological material. The present study aimed at optimizing the extraction of C-phycocyanin (C-PC) and other water-soluble biomolecules from <em>Arthrospira platensis</em> cyanobacterium by adjusting PEF process parameters. At a field strength of 5 kV/cm, specific energies comprised between 2.1 and 41.1 MJ/kg_DW were applied to the PEF treatment chamber containing an aqueous solution with 1.11 g_DW/kg_SUS of <em>A. platensis.</em> Experimental results showed that extraction efficiency including C-PC yield and quality are strongly influenced by PEF specific energy. The application of 20.5 MJ/kg_DW generated a C-PC yield of 84 mg_C-PC/g_DW with purity of 0.52 (A₆₂₀/A₂₈₀) and selectivity of 3.10 (A₆₂₀/A₆₈₀), whereas only 56 % of the cells were disintegrated. Higher specific energies of 30.8 MJ/kg_DW improved the disruption of <em>A. platensis</em> cells (85 %) and the C-PC extraction yield (115 mg_C-PC/g_DW) but at the expense of extracts quality. This study highlights the importance of finding a compromise between PEF energy requirements and extraction performances, which can have significant impact on the overall economic viability of <em>A. platensis</em> downstream processes.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"407 ","pages":"Pages 59-69"},"PeriodicalIF":3.9,"publicationDate":"2025-08-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144812050","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Utilizing the SacB-mediated gene editing system in Komagataeibacter xylinus to explore the function of bacterial cellulose synthase","authors":"Zhaojun Peng ,&nbsp;Xinyue Gan ,&nbsp;Jiaheng Liu ,&nbsp;Bo Xin ,&nbsp;Cheng Zhong","doi":"10.1016/j.jbiotec.2025.08.003","DOIUrl":"10.1016/j.jbiotec.2025.08.003","url":null,"abstract":"<div><div>Bacterial cellulose (BC) is a microbial polysaccharide, which is widely used in biotechnology, food, medicine, and other industries. Although existing genetic toolkits have laid a solid foundation for the genetic manipulation of BC-producing strains, there is still room for improvement in enhancing editing efficiency, simplifying operational procedures, and achieving scarless modifications. In the present study, we developed a SacB-based system, pK18mobsacB, to achieve marker-free gene editing with an efficiency of up to 83.33 %. Gene deletion, insertion, and replacement were successfully performed in <em>Komagataeibacter xylinus</em> CGMCC 2955 using this system. Subsequently, the SacB-based system was used to explore the function of bacterial cellulose synthase in the synthesis and structure of BC. It was found that the <em>bcs I</em> operon played an important role in BC synthesis. The deletion of the <em>bcs II</em> and <em>bcs III</em> operon regions, either individually or in combination, led to an increase in the fiber diameter and crystallinity of the BC films. The SacB-based system and its applications established in this study provide valuable tools and a theoretical foundation for the modification of BC-producing strains using synthetic biology, thereby facilitating the sustainable application of BC and the development of innovative products.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"407 ","pages":"Pages 48-58"},"PeriodicalIF":3.9,"publicationDate":"2025-08-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144799176","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Microbial production of single cell oil from biowaste-derived volatile fatty acids using a newly identified oleaginous Cyberlindnera jadinii","authors":"Ticiana Fernandes ,&nbsp;Pier Parpot ,&nbsp;Isabel Soares-Silva ,&nbsp;Maria João Sousa ,&nbsp;Ricardo Franco-Duarte","doi":"10.1016/j.jbiotec.2025.08.004","DOIUrl":"10.1016/j.jbiotec.2025.08.004","url":null,"abstract":"<div><div>Efforts to achieve a circular economy emphasize waste minimization while maximizing its value through recycling and reuse. Volatile Fatty Acids (VFAs), derived from organic waste streams, hold biotechnological potential as raw materials for producing value-added compounds offering sustainable alternatives to traditional petroleum-based approaches. The phenotypic and genomic plasticity of non-<em>Saccharomyces</em> yeasts positions them as ideal candidates for VFAs metabolization. In this study, the growth profiles of 48 yeasts were assessed using seven VFAs as sole carbon sources, as representative of a real organic waste stream composition. Growth was assessed individually and in mixtures of the VFAs, at concentrations representative of the organic biowastes. Using AMiGA tool we deepened yeasts growth dynamics based on the optical density growth curves measurements. Ten isolates showcased superior performance, through their varied growth under different VFAs combinations. Acetic and isobutyric acids were the most favourable VFAs for non-<em>Saccharomyces</em> yeasts growth, balancing efficiency and adaptability. In contrast, butyric and caproic acids were the less effective, likely due to metabolic constraints or inhibitory effects. Yeasts favoured lipid synthesis over TCA intermediates, glycerol and ethanol. Two <em>C. jadinii</em> strains stood out as particularly interesting considering their metabolic plasticity, successfully assimilating both individual and combined VFAs, achieving the highest growth coupled with a superior lipid production (19.5–26.7 % (w/w)). These findings establish <em>C. jadinii</em> as an oleaginous yeast, a trait firstly reported here, underscoring its promise as a cornerstone for biowaste valorisation.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"407 ","pages":"Pages 31-42"},"PeriodicalIF":3.9,"publicationDate":"2025-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144799175","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Stepwise metabolic engineering of a plasmid-free Corynebacterium glutamicum for efficient production of γ-aminobutyric acid (GABA) by co-utilizing lignocellulosic feedstock-derived sugars","authors":"Jie Wang,&nbsp;Yingying Xu,&nbsp;Zhuolin Song,&nbsp;Bin Zhang,&nbsp;Jie Bao","doi":"10.1016/j.jbiotec.2025.07.023","DOIUrl":"10.1016/j.jbiotec.2025.07.023","url":null,"abstract":"<div><div>γ-aminobutyric acid (GABA) can be synthesized through plasmid-based expression of glutamate decarboxylase in L-glutamic acid producing <em>Corynebacterium glutamicum</em> strain. However, the addition of antibiotic to maintain the expression plasmid during the fermentation not only increases production and recovery costs, but also poses potential food safety hazards. In this study, a plasmid-free GABA producing <em>C. glutamicum</em> strain was constructed from <em>C. glutamicum</em> GJ04 chassis, which can produce L-glutamate by co-utilizing lignocellulose-derived glucose and xylose. Secretory glutamate decarboxylase was integrated into the genome of <em>C. glutamicum</em> GJ04 in three copies by replacing <em>ldh</em>, <em>gabT</em>, <em>gabD</em> genes. The metabolic flux in engineered <em>C. glutamicum</em> was further fine-tuned by knocking out <em>aceA</em> and <em>gabP</em> genes to enhance GABA production. The recombinant strain <em>C. glutamicum</em> GJ09 can produce 44.3 ± 3.8 g/L GABA from 15 % (w/w) solids loading corncob residues hydrolysate with the yield and productivity of 0.45 g/g and 0.74 g/L/h. The highest GABA titer reached 63.4 g/L by fed-batch fermentation using corncob residues-derived syrup. This study provided a robust and plasmid-free <em>C. glutamicum</em> strain by stepwise metabolic engineering for industrial production of GABA from lignocellulosic feedstocks.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"406 ","pages":"Pages 281-284"},"PeriodicalIF":3.9,"publicationDate":"2025-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144724519","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The many virtues of staphylococcal protein A: A journey from N to C terminus","authors":"Charles Dahlsson Leitao,&nbsp;John Löfblom,&nbsp;Per-Åke Nygren,&nbsp;Sophia Hober,&nbsp;Mathias Uhlén,&nbsp;Stefan Ståhl","doi":"10.1016/j.jbiotec.2025.07.018","DOIUrl":"10.1016/j.jbiotec.2025.07.018","url":null,"abstract":"<div><div>This review outlines the historical development and versatile applications of one of the most well-studied bacterial proteins, namely the immunoglobulin (Ig)-binding staphylococcal protein A (SpA) of <em>Staphylococcus aureus</em>. Each segment of the SpA operon, from the 5’ promoter region and signal peptide to the 3’ cell wall anchoring region, has been exploited for various innovative applications in areas such as immunology and biotechnology. We provide an overview of selected applications and concepts that have had a significant impact on life science research, and some that have also led to significant commercial implications. In the 1980s, the SpA promoter and signal sequence were utilized in <em>Escherichia coli</em> for recombinant production of various proteins, yielding product secretion to the culture medium and thereby simplifying product recovery. The five homologous Ig-binding domains of SpA gained tremendous interest in the late 1980s, largely due to the rise of monoclonal antibodies (mAbs) for therapeutic use, prompting a growing demand for effective affinity ligands to facilitate their purification. Over the years, these Ig-binding domains have been extensively investigated and re-engineered to bind proteins other than antibodies, leading in the mid-1990s to the development of the affibody affinity protein technology. Today, affibody molecules are being investigated in late-stage clinical trials as potential protein therapeutics for various indications. Finally, the cell wall anchoring regions of SpA inspired the development of a surface display system for <em>Staphylococcus carnosus</em>, which has emerged as a technology platform in combinatorial protein engineering for work with large peptide, antibody and affibody libraries.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"406 ","pages":"Pages 272-280"},"PeriodicalIF":3.9,"publicationDate":"2025-07-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144724681","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}