Pub Date : 2025-07-29DOI: 10.1016/j.btre.2025.e00908
Pongtorn Phengnoi , Nuttinee Teerakulkittipong , Kosin Teeparuksapun , Gary Antonio Lirio , Witawat Jangiam
Levan, a promising fructan polysaccharide for biopharmaceuticals, has limited large-scale production studies. This research optimized and scaled up levan biosynthesis from Bacillus siamensis in continuous stirred-tank bioreactors based on response surface methodology (RSM). Batch cultures optimized for 30 % (w/v) sucrose, pH 5.0, and 48 h incubation yielded a maximum 15.74 % (w/v) levan. The optimal batch conditions were evaluated in a continuous stirred-tank bioreactor, where dilution rates and mixing speeds were examined. At a dilution rate of 0.021 h⁻¹ and an agitation speed of 200 rpm, the maximum productivity was 17.96 % (w/v), and steady-state conditions were attained after three days of continuous fermentation. X-ray diffraction confirmed the amorphous nature of the levan, ideal for biomaterial applications. These results underline the potential of B. siamensis for high-yield levan production and provide a systematic approach for bioprocess parameter optimization, serving as a strong basis for its increased application in industrialized polysaccharide-based bioprocessing.
{"title":"Optimization of levan biosynthesis from Bacillus siamensis using batch and continuous stirred-tank bioreactors: A response surface methodology approach","authors":"Pongtorn Phengnoi , Nuttinee Teerakulkittipong , Kosin Teeparuksapun , Gary Antonio Lirio , Witawat Jangiam","doi":"10.1016/j.btre.2025.e00908","DOIUrl":"10.1016/j.btre.2025.e00908","url":null,"abstract":"<div><div>Levan, a promising fructan polysaccharide for biopharmaceuticals, has limited large-scale production studies. This research optimized and scaled up levan biosynthesis from <em>Bacillus siamensis</em> in continuous stirred-tank bioreactors based on response surface methodology (RSM). Batch cultures optimized for 30 % (w/v) sucrose, pH 5.0, and 48 h incubation yielded a maximum 15.74 % (w/v) levan. The optimal batch conditions were evaluated in a continuous stirred-tank bioreactor, where dilution rates and mixing speeds were examined. At a dilution rate of 0.021 h⁻¹ and an agitation speed of 200 rpm, the maximum productivity was 17.96 % (w/v), and steady-state conditions were attained after three days of continuous fermentation. X-ray diffraction confirmed the amorphous nature of the levan, ideal for biomaterial applications. These results underline the potential of <em>B. siamensis</em> for high-yield levan production and provide a systematic approach for bioprocess parameter optimization, serving as a strong basis for its increased application in industrialized polysaccharide-based bioprocessing.</div></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"47 ","pages":"Article e00908"},"PeriodicalIF":0.0,"publicationDate":"2025-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144771035","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-07-22DOI: 10.1016/j.btre.2025.e00905
Marwan Taha , Anna-Maria Pappa , Hani Saleh , Anas Alazzam
This paper examines recent advancements in cell characterization using microfluidic devices, emphasizing mechanical, electrical, and hybrid methodologies. These technologies have substantially improved throughput, precision, and the range of cell types they can analyze. Key microfluidic technologies for cell characterization are reviewed, including label-free electrical and mechanical methods designed for high-throughput, real-time analysis. Microfluidic advancements in cell characterization are critically assessed, along with challenges such as operational complexity and the need for more adaptable, user-friendly platforms. The integration of AI and machine learning in microfluidic systems is also discussed, highlighting their crucial role in automating data analysis and enhancing classification accuracy, with implications for personalized medicine and advanced cellular assays.
{"title":"Enhancing cell characterization with microfluidics and AI: a comprehensive review of mechanical, electrical, and hybrid techniques","authors":"Marwan Taha , Anna-Maria Pappa , Hani Saleh , Anas Alazzam","doi":"10.1016/j.btre.2025.e00905","DOIUrl":"10.1016/j.btre.2025.e00905","url":null,"abstract":"<div><div>This paper examines recent advancements in cell characterization using microfluidic devices, emphasizing mechanical, electrical, and hybrid methodologies. These technologies have substantially improved throughput, precision, and the range of cell types they can analyze. Key microfluidic technologies for cell characterization are reviewed, including label-free electrical and mechanical methods designed for high-throughput, real-time analysis. Microfluidic advancements in cell characterization are critically assessed, along with challenges such as operational complexity and the need for more adaptable, user-friendly platforms. The integration of AI and machine learning in microfluidic systems is also discussed, highlighting their crucial role in automating data analysis and enhancing classification accuracy, with implications for personalized medicine and advanced cellular assays.</div></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"47 ","pages":"Article e00905"},"PeriodicalIF":0.0,"publicationDate":"2025-07-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144678845","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-07-17DOI: 10.1016/j.btre.2025.e00906
Silvia Abdi Pratama , Adi Setyo Purnomo , Asranudin Asranudin
Methylene blue (MB) is a common synthetic dye used in various industries due to its abundance and cost-effectiveness. However, the presence of this synthetic dye in industrial wastewater has the potential to cause significant effects on the environment and public health when released into soil or water bodies. According to results, biological wastewater treatment has emerged as a promising approach due to its economic feasibility, efficiency, and environmental compatibility. Therefore, this study aims to evaluate the MB removal by Pseudomonas aeruginosa immobilized within alginate–polyvinyl alcohol (PVA) matrices and compare removal performance to that of free (non-immobilized) cells. The results showed that the immobilized P. aeruginosa (Alg-PVA-PA beads) exhibited higher MB removal efficiency (72.52%) compared to free cells (55.52%) and Alg-PVA beads alone (43.12%). Adsorption analysis showed that the removal process followed a pseudo-second-order kinetic model, suggesting chemisorption, and was best described by the Langmuir isotherm, showing monolayer adsorption. Additionally, reusability tests showed that the immobilized biocomposite beads retained over 35% MB removal efficiency after five consecutive cycles, suggesting the sustainability of the beads. XRD, FTIR, and SEM-EDX analysis confirmed the successful entrapment of P. aeruginosa within the Alg-PVA matrices and showed structural changes in the beads following the MB removal process. The results were also supported by LC-QTOF/MS outcomes, which showed metabolites with retention times of 1.17, 1.52, 8.35, and 9.36 min, suggesting the successful degradation of MB.
{"title":"Methylene Blue Removal Using Alginate-PVA-Pseudomonas aeruginosa Biocomposite: Kinetics and Biodegradation Studies","authors":"Silvia Abdi Pratama , Adi Setyo Purnomo , Asranudin Asranudin","doi":"10.1016/j.btre.2025.e00906","DOIUrl":"10.1016/j.btre.2025.e00906","url":null,"abstract":"<div><div>Methylene blue (MB) is a common synthetic dye used in various industries due to its abundance and cost-effectiveness. However, the presence of this synthetic dye in industrial wastewater has the potential to cause significant effects on the environment and public health when released into soil or water bodies. According to results, biological wastewater treatment has emerged as a promising approach due to its economic feasibility, efficiency, and environmental compatibility. Therefore, this study aims to evaluate the MB removal by <em>Pseudomonas aeruginosa</em> immobilized within alginate–polyvinyl alcohol (PVA) matrices and compare removal performance to that of free (non-immobilized) cells. The results showed that the immobilized <em>P. aeruginosa</em> (Alg-PVA-PA beads) exhibited higher MB removal efficiency (72.52%) compared to free cells (55.52%) and Alg-PVA beads alone (43.12%). Adsorption analysis showed that the removal process followed a pseudo-second-order kinetic model, suggesting chemisorption, and was best described by the Langmuir isotherm, showing monolayer adsorption. Additionally, reusability tests showed that the immobilized biocomposite beads retained over 35% MB removal efficiency after five consecutive cycles, suggesting the sustainability of the beads. XRD, FTIR, and SEM-EDX analysis confirmed the successful entrapment of <em>P. aeruginosa</em> within the Alg-PVA matrices and showed structural changes in the beads following the MB removal process. The results were also supported by LC-QTOF/MS outcomes, which showed metabolites with retention times of 1.17, 1.52, 8.35, and 9.36 min, suggesting the successful degradation of MB.</div></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"47 ","pages":"Article e00906"},"PeriodicalIF":0.0,"publicationDate":"2025-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144738918","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-07-16DOI: 10.1016/j.btre.2025.e00907
Maryam Anar , Sajjad Kamal Shuvro , Muhammad Farooq Hussain Munis , Masaaki Morikawa
The effects of biologically synthesized MnO and ZnO microparticles (MPs), were evaluated on the growth of Lemna minor. Both MnO and ZnO MPs promoted the growth of L. minor at low concentrations of 1–8 mg/L. In contrast, 1000 mg/L MnO and ZnO MPs reduced the growth of L. minor. It was also found that the starch content of L. minor and Spirodela polyrhiza was enhanced upon growing in 1 mg/L of MnO MPs when compared to no MnO MPs. These effects were suggested to be due in part to the fact that biogenic MPs harbored small but significant amounts of inorganic phosphorus. Our findings indicate that biologically synthesized MnO and ZnO MPs can act as novel plant growth regulators to enhance the production of useful duckweed biomass with high starch content.
{"title":"Enhancement of the duckweed biomass and starch production utilizing biogenic MnO and ZnO microparticles","authors":"Maryam Anar , Sajjad Kamal Shuvro , Muhammad Farooq Hussain Munis , Masaaki Morikawa","doi":"10.1016/j.btre.2025.e00907","DOIUrl":"10.1016/j.btre.2025.e00907","url":null,"abstract":"<div><div>The effects of biologically synthesized MnO and ZnO microparticles (MPs), were evaluated on the growth of <em>Lemna minor</em>. Both MnO and ZnO MPs promoted the growth of L. <em>minor</em> at low concentrations of 1–8 mg/L. In contrast, 1000 mg/L MnO and ZnO MPs reduced the growth of L. <em>minor</em>. It was also found that the starch content of L. <em>minor</em> and <em>Spirodela polyrhiza</em> was enhanced upon growing in 1 mg/L of MnO MPs when compared to no MnO MPs. These effects were suggested to be due in part to the fact that biogenic MPs harbored small but significant amounts of inorganic phosphorus. Our findings indicate that biologically synthesized MnO and ZnO MPs can act as novel plant growth regulators to enhance the production of useful duckweed biomass with high starch content.</div></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"47 ","pages":"Article e00907"},"PeriodicalIF":0.0,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144704543","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-06-30DOI: 10.1016/j.btre.2025.e00904
Tímea B. Gerzsenyi , Ágnes M. Ilosvai , Ferenc Kristály , Lajos Daróczi , Michael C. Owen , Béla Viskolcz , László Vanyorek , Emma Szőri-Dorogházi
DNA isolation is a crucial step in many molecular biological applications for diagnostic and research purposes, like detection of infectious diseases or gene expression studies. However, due to the requirement of toxic reagents in traditional procedures and the high expenses of commercial kits, the use of magnetic MNP-based DNA isolation is becoming more widespread. In this study, different ferrite containing MNPs (MnFe2O4, MnFe2O4-NH2, MgFe2O4, MgFe2O4-NH2 NiFe2O4, NiFe2O4-NH2) are examined and compared in their pDNA isolation efficiency. Among the tested nanoparticles, we document the use of NiFe2O4 and its amine-functionalized form for the first time. Three protocols for the isolation of pDNA are optimized for each type of nanoparticle and the best protocol is selected based on the quantity, quality and integrity of the extracted DNA. Plasmid samples extracted with the MNPs are transformed into competent bacterial cells and further tests are performed to recover genomic DNA from bacterial cells, leading to the development of another protocol. Bacteria-spiked blood serum samples are produced to extract DNA from a more complex biological matrix.
{"title":"Investigation and optimization of DNA isolation efficiency using ferrite-based magnetic nanoparticles","authors":"Tímea B. Gerzsenyi , Ágnes M. Ilosvai , Ferenc Kristály , Lajos Daróczi , Michael C. Owen , Béla Viskolcz , László Vanyorek , Emma Szőri-Dorogházi","doi":"10.1016/j.btre.2025.e00904","DOIUrl":"10.1016/j.btre.2025.e00904","url":null,"abstract":"<div><div>DNA isolation is a crucial step in many molecular biological applications for diagnostic and research purposes, like detection of infectious diseases or gene expression studies. However, due to the requirement of toxic reagents in traditional procedures and the high expenses of commercial kits, the use of magnetic MNP-based DNA isolation is becoming more widespread. In this study, different ferrite containing MNPs (MnFe<sub>2</sub>O<sub>4</sub>, MnFe<sub>2</sub>O<sub>4</sub>-NH<sub>2</sub>, MgFe<sub>2</sub>O<sub>4</sub>, MgFe<sub>2</sub>O<sub>4</sub>-NH<sub>2</sub> NiFe<sub>2</sub>O<sub>4</sub>, NiFe<sub>2</sub>O<sub>4</sub>-NH<sub>2</sub>) are examined and compared in their pDNA isolation efficiency. Among the tested nanoparticles, we document the use of NiFe<sub>2</sub>O<sub>4</sub> and its amine-functionalized form for the first time. Three protocols for the isolation of pDNA are optimized for each type of nanoparticle and the best protocol is selected based on the quantity, quality and integrity of the extracted DNA. Plasmid samples extracted with the MNPs are transformed into competent bacterial cells and further tests are performed to recover genomic DNA from bacterial cells, leading to the development of another protocol. Bacteria-spiked blood serum samples are produced to extract DNA from a more complex biological matrix.</div></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"47 ","pages":"Article e00904"},"PeriodicalIF":0.0,"publicationDate":"2025-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144571879","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-06-24DOI: 10.1016/j.btre.2025.e00903
María Clara De La Hoz-Romo , Luis Díaz , Javier Gómez-León , Luis Felipe Reyes , Marynes Quintero , Luisa Villamil
Quorum sensing (QS) regulates virulence factors in bacteria associated with acne vulgaris. Consequently, extracts from marine actinobacteria sourced from Colombian Caribbean sponges and octocorals were screened for quorum quenching (QQ) and anti-biofilm activity. The most promising extract, G12.218 from Promicromonospora sp. CLIVUS-G12218, inhibited QS by 97 % via AI-2 signaling in Vibrio harveyi BB170 and reduced Staphylococcus aureus biofilm by 60 % at 3 mg/L, without cytotoxicity to keratinocytes and fibroblasts.
Untargeted metabolomics of QQ active and non-active extracts revealed distinct profiles enriched in terpenoids, phenols, and butenolides, potentially linked to bioactivity. These extracts also exhibited antioxidant capacity (18 ± 0.23 µmol TE/L) and hyaluronidase inhibition up to 72.8 %, suggesting skin-protective properties. These findings highlight the potential of marine actinobacteria as a source of QQ and anti-biofilm agents for acne therapy and skincare.
{"title":"Quorum quenching: A key biological activity of marine actinobacteria extracts for acne vulgaris control","authors":"María Clara De La Hoz-Romo , Luis Díaz , Javier Gómez-León , Luis Felipe Reyes , Marynes Quintero , Luisa Villamil","doi":"10.1016/j.btre.2025.e00903","DOIUrl":"10.1016/j.btre.2025.e00903","url":null,"abstract":"<div><div>Quorum sensing (QS) regulates virulence factors in bacteria associated with acne vulgaris. Consequently, extracts from marine actinobacteria sourced from Colombian Caribbean sponges and octocorals were screened for quorum quenching (QQ) and anti-biofilm activity. The most promising extract, G12.218 from <em>Promicromonospora</em> sp. CLIVUS-G12218, inhibited QS by 97 % via AI-2 signaling in <em>Vibrio harveyi</em> BB170 and reduced <em>Staphylococcus aureus</em> biofilm by 60 % at 3 mg/L, without cytotoxicity to keratinocytes and fibroblasts.</div><div>Untargeted metabolomics of QQ active and non-active extracts revealed distinct profiles enriched in terpenoids, phenols, and butenolides, potentially linked to bioactivity. These extracts also exhibited antioxidant capacity (18 ± 0.23 µmol TE/L) and hyaluronidase inhibition up to 72.8 %, suggesting skin-protective properties. These findings highlight the potential of marine actinobacteria as a source of QQ and anti-biofilm agents for acne therapy and skincare.</div></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"47 ","pages":"Article e00903"},"PeriodicalIF":0.0,"publicationDate":"2025-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144858079","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-06-23DOI: 10.1016/j.btre.2025.e00902
Loretta László , Anna Lovrics , Álmos Tilajka , Tamás Takács , László Buday , Virag Vas
Cancer rarely results from a single gene defect but emerges from disruptions in complex cellular networks. The Network Medicine perspective guides our investigation of cancer-driving interactions, particularly focusing on RAS signaling pathways that are key mediator for cancer development.
We analyzed gene expression patterns in colon and lung cancers to identify stage-specific molecular drivers. Using computational modelling combined with patient tissue analysis, we discovered five key genes that are specifically altered in early-stage colon cancer: RAF1, PLCE1, RGL1, RIN1, and GRB7. These genes work as RAS effectors in signaling and can effectively distinguish between normal and cancerous colon tissue.
Our approach combines network analysis with gene expression studies to understand how RAS signaling disruption contributes to colon cancer development. These findings suggest that targeting early-stage RAS-related changes could offer therapeutic opportunities before cancer becomes more complex and harder to treat.
{"title":"Integrative analysis of RAS signaling effectors reveals stage-dependent oncogenic patterns in colon adenocarcinoma","authors":"Loretta László , Anna Lovrics , Álmos Tilajka , Tamás Takács , László Buday , Virag Vas","doi":"10.1016/j.btre.2025.e00902","DOIUrl":"10.1016/j.btre.2025.e00902","url":null,"abstract":"<div><div>Cancer rarely results from a single gene defect but emerges from disruptions in complex cellular networks. The Network Medicine perspective guides our investigation of cancer-driving interactions, particularly focusing on RAS signaling pathways that are key mediator for cancer development.</div><div>We analyzed gene expression patterns in colon and lung cancers to identify stage-specific molecular drivers. Using computational modelling combined with patient tissue analysis, we discovered five key genes that are specifically altered in early-stage colon cancer: RAF1, PLCE1, RGL1, RIN1, and GRB7. These genes work as RAS effectors in signaling and can effectively distinguish between normal and cancerous colon tissue.</div><div>Our approach combines network analysis with gene expression studies to understand how RAS signaling disruption contributes to colon cancer development. These findings suggest that targeting early-stage RAS-related changes could offer therapeutic opportunities before cancer becomes more complex and harder to treat.</div></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"47 ","pages":"Article e00902"},"PeriodicalIF":0.0,"publicationDate":"2025-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144510914","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-05-22DOI: 10.1016/j.btre.2025.e00900
James Colter , Tiffany Dang , Julia Malinovska , Jessica May Corpuz , Dora Modrcin , Roman Krawetz , Kartikeya Murari , Michael Scott Kallos
Human induced pluripotent stem cell (hiPSC) derived therapeutics require clinically relevant quantities of high-quality cell populations for applications in regenerative medicine. The lack of efficacy exhibited across clinical trials suggests deeper understanding of the networks governing phenotype is needed. Further, costs limit study throughput in characterizing the artificial niche relative to outcomes. We present herein an optimized strategy to enable high-throughput hiPSC expansion at <20 mL research scale. We assessed viability of single cell inoculation and aggregate preformation to facilitate proliferation. We modeled aggregate characteristics against agitation rate. Our results demonstrate tunable control with fold expansion comparable to commercial systems. Marker quantification and teratoma assay confirm functional pluripotency. This approach constitutes a scalable protocol to accelerate hiPSC research, and a significant step in advancing the rate of progress in elucidating links to derivative functionality. This work will enable statistically rigorous studies targeting hiPSC and downstream phenotype for clinical manufacturing.
{"title":"Scale-down optimization of a robust, parallelizable human induced pluripotent stem cell bioprocess for high-throughput research","authors":"James Colter , Tiffany Dang , Julia Malinovska , Jessica May Corpuz , Dora Modrcin , Roman Krawetz , Kartikeya Murari , Michael Scott Kallos","doi":"10.1016/j.btre.2025.e00900","DOIUrl":"10.1016/j.btre.2025.e00900","url":null,"abstract":"<div><div>Human induced pluripotent stem cell (hiPSC) derived therapeutics require clinically relevant quantities of high-quality cell populations for applications in regenerative medicine. The lack of efficacy exhibited across clinical trials suggests deeper understanding of the networks governing phenotype is needed. Further, costs limit study throughput in characterizing the artificial niche relative to outcomes. We present herein an optimized strategy to enable high-throughput hiPSC expansion at <20 mL research scale. We assessed viability of single cell inoculation and aggregate preformation to facilitate proliferation. We modeled aggregate characteristics against agitation rate. Our results demonstrate tunable control with fold expansion comparable to commercial systems. Marker quantification and teratoma assay confirm functional pluripotency. This approach constitutes a scalable protocol to accelerate hiPSC research, and a significant step in advancing the rate of progress in elucidating links to derivative functionality. This work will enable statistically rigorous studies targeting hiPSC and downstream phenotype for clinical manufacturing.</div></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"47 ","pages":"Article e00900"},"PeriodicalIF":0.0,"publicationDate":"2025-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144168992","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-05-20DOI: 10.1016/j.btre.2025.e00899
Halima Aliyu Alhafiz , Karin Longus , Rob A.J. Verlinden , Vera Lambauer , Andreas Kruschitz , Regina Kratzer
Today, 85 % of the carbon in organic chemicals and their derivatives comes from fossil sources. Replacing fossil-based materials with sustainable sources requires large quantities of feedstocks and mature technologies. Biorefineries based on lignocellulose have great potential to replace fossil raw materials in the short and medium term. Here we want to pave the way for the bacterium Cupriavidus necator as a versatile biotechnological workhorse in future biorefineries. Wheat straw, beech, pine and spruce reflect lignocellulosic biomass from the agricultural waste and wood sectors that is widespread in Europe. Miscanthus was chosen as an emerging energy crop. Lignocellulose feedstocks were pretreated by steam explosion under variable conditions prior to enzymatic hydrolysis. Native Cupriavidus necator and a strain adapted by laboratory evolution were shown to grow on 16 filtered lignocellulosic hydrolysates as the sole carbon source and without prior detoxification.
{"title":"Cultivation of Cupriavidus necatorstrains on hydrolyzed lignocellulosic feedstocks widely available in Europe","authors":"Halima Aliyu Alhafiz , Karin Longus , Rob A.J. Verlinden , Vera Lambauer , Andreas Kruschitz , Regina Kratzer","doi":"10.1016/j.btre.2025.e00899","DOIUrl":"10.1016/j.btre.2025.e00899","url":null,"abstract":"<div><div>Today, 85 % of the carbon in organic chemicals and their derivatives comes from fossil sources. Replacing fossil-based materials with sustainable sources requires large quantities of feedstocks and mature technologies. Biorefineries based on lignocellulose have great potential to replace fossil raw materials in the short and medium term. Here we want to pave the way for the bacterium <em>Cupriavidus necator</em> as a versatile biotechnological workhorse in future biorefineries. Wheat straw, beech, pine and spruce reflect lignocellulosic biomass from the agricultural waste and wood sectors that is widespread in Europe. Miscanthus was chosen as an emerging energy crop. Lignocellulose feedstocks were pretreated by steam explosion under variable conditions prior to enzymatic hydrolysis. Native <em>Cupriavidus necator</em> and a strain adapted by laboratory evolution were shown to grow on 16 filtered lignocellulosic hydrolysates as the sole carbon source and without prior detoxification.</div></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"47 ","pages":"Article e00899"},"PeriodicalIF":0.0,"publicationDate":"2025-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144240520","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Proinsulin was expressed by Escherichia coli SHuffle T7 with pET system in minimal medium containing glucose (Glu medium), glucose and glycerol (GluGly medium) or glycerol (Gly medium). With 100 μM IPTG, proinsulin production did not increase with glycerol. In contrast, proinsulin production per medium volume in GluGly and Gly media was approximately 3∼4-fold higher than in Glu medium with 10 μM IPTG. mRNA expression of target protein was higher in GluGly versus Glu medium, indicating that proinsulin production was enhanced by the release of glucose-induced catabolite inhibition. Although proinsulin production did not differ between GluGly and Gly media at 25 h, substrate was consumed quickly in GluGly medium with 1.55±0.12 times higher proinsulin production at 15 h. Productivity, considering the production period, was highest in the GluGly medium. This study shows a mixture of glucose and glycerol is valuable for protein production in E. coli with low IPTG concentrations.
{"title":"Induction conditions that promote the effect of glycerol on recombinant protein production in Escherichia coli","authors":"Yoshihiro Ojima , Hajime Saito , Shintaro Miyuki , Koichi Fukunaga , Terumichi Tsuboi , Masayuki Azuma","doi":"10.1016/j.btre.2025.e00898","DOIUrl":"10.1016/j.btre.2025.e00898","url":null,"abstract":"<div><div>Proinsulin was expressed by <em>Escherichia coli</em> SHuffle T7 with pET system in minimal medium containing glucose (Glu medium), glucose and glycerol (GluGly medium) or glycerol (Gly medium). With 100 μM IPTG, proinsulin production did not increase with glycerol. In contrast, proinsulin production per medium volume in GluGly and Gly media was approximately 3∼4-fold higher than in Glu medium with 10 μM IPTG. mRNA expression of target protein was higher in GluGly versus Glu medium, indicating that proinsulin production was enhanced by the release of glucose-induced catabolite inhibition. Although proinsulin production did not differ between GluGly and Gly media at 25 h, substrate was consumed quickly in GluGly medium with 1.55±0.12 times higher proinsulin production at 15 h. Productivity, considering the production period, was highest in the GluGly medium. This study shows a mixture of glucose and glycerol is valuable for protein production in <em>E. coli</em> with low IPTG concentrations.</div></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"46 ","pages":"Article e00898"},"PeriodicalIF":0.0,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144124178","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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