Pub Date : 2026-01-19DOI: 10.1186/s13036-025-00604-x
Marisabel Morales-Muralles, Spencer J Serrano, George Makhatadze, José I Reyes-De-Corcuera
We report the effects of high hydrostatic pressure (HHP) on D-amino acid oxidase (DAAO) from pork kidney stability and activity. At 20 °C, the activity decreased with increasing pressure, but at 40 °C, it was not significantly affected because the increased catalytic rate compensated for the enzyme inactivation within the time frame of the experiment. At 50-300 MPa, raising the temperature increased the rate of DAAO inactivation, indicating that increases in both pressure and temperature contributed to inactivation. In contrast to other oxidases pressure did not stabilize DAAO against thermal inactivation. Treatments between 0.1 and 100 MPa had no significant effect on its stability at 55 °C. However, the rate of inactivation increased with increasing pressure from 100 to 300 MPa, resulting in an activation volume (ΔV‡) of inactivation of -15.8 ± 1.7 cm³mol⁻¹. Unlike other oxidases that have buried cavities larger than 100 Å3 (0.1 nm3), DAAO has cavities smaller than 75 Å3 (0.075 nm3), thus supporting the hypothesis that cavity size determines whether pressure stabilizes an enzyme against thermal inactivation.
{"title":"Effect of high hydrostatic pressure on D-amino acid oxidase activity and stability.","authors":"Marisabel Morales-Muralles, Spencer J Serrano, George Makhatadze, José I Reyes-De-Corcuera","doi":"10.1186/s13036-025-00604-x","DOIUrl":"https://doi.org/10.1186/s13036-025-00604-x","url":null,"abstract":"<p><p>We report the effects of high hydrostatic pressure (HHP) on D-amino acid oxidase (DAAO) from pork kidney stability and activity. At 20 °C, the activity decreased with increasing pressure, but at 40 °C, it was not significantly affected because the increased catalytic rate compensated for the enzyme inactivation within the time frame of the experiment. At 50-300 MPa, raising the temperature increased the rate of DAAO inactivation, indicating that increases in both pressure and temperature contributed to inactivation. In contrast to other oxidases pressure did not stabilize DAAO against thermal inactivation. Treatments between 0.1 and 100 MPa had no significant effect on its stability at 55 °C. However, the rate of inactivation increased with increasing pressure from 100 to 300 MPa, resulting in an activation volume (ΔV<sup>‡</sup>) of inactivation of -15.8 ± 1.7 cm³mol⁻¹. Unlike other oxidases that have buried cavities larger than 100 Å<sup>3</sup> (0.1 nm<sup>3</sup>), DAAO has cavities smaller than 75 Å<sup>3</sup> (0.075 nm<sup>3</sup>), thus supporting the hypothesis that cavity size determines whether pressure stabilizes an enzyme against thermal inactivation.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":""},"PeriodicalIF":6.5,"publicationDate":"2026-01-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146003556","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}
Pub Date : 2026-01-10DOI: 10.1186/s13036-025-00607-8
Mahsa Mohammadzadeh, Ali Farzin, Zahra Pazhouhnia, Mahdieh Hoseinpour, Nima Beheshtizadeh
{"title":"Smart biomaterials for cardiovascular, bone, and skin tissue engineering: mechanisms, applications, and future prospects.","authors":"Mahsa Mohammadzadeh, Ali Farzin, Zahra Pazhouhnia, Mahdieh Hoseinpour, Nima Beheshtizadeh","doi":"10.1186/s13036-025-00607-8","DOIUrl":"10.1186/s13036-025-00607-8","url":null,"abstract":"","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":"31"},"PeriodicalIF":6.5,"publicationDate":"2026-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145948867","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}
Pub Date : 2026-01-09DOI: 10.1186/s13036-025-00588-8
Ignas Lebedis, Jolita Pachaleva, Eiva Bernotiene, Daiva Bironaite, Tomas Ragauskas, Giedrius Kvedaras, Gunaras Terbetas, Ilona Uzieliene
{"title":"Effect of HIF-1α inhibitor LW6 on chondrogenic properties of mesenchymal stromal cells and chondrocytes in cell sheets under physioxia.","authors":"Ignas Lebedis, Jolita Pachaleva, Eiva Bernotiene, Daiva Bironaite, Tomas Ragauskas, Giedrius Kvedaras, Gunaras Terbetas, Ilona Uzieliene","doi":"10.1186/s13036-025-00588-8","DOIUrl":"10.1186/s13036-025-00588-8","url":null,"abstract":"","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":"30"},"PeriodicalIF":6.5,"publicationDate":"2026-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145943984","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}
Pub Date : 2026-01-08DOI: 10.1186/s13036-025-00611-y
Danny Knobloch-Sperlich, Matthias Kappler, Markus Glaß, Antje Güttler, Marina Petrenko, Jonas Pyko, Tony Gutschner, Frank Tavasol, Dirk Vordermark, Matthias Bache
{"title":"Two-color spheroid model for determining the O<sub>2</sub>-induced radiosensitivity of HNSCC.","authors":"Danny Knobloch-Sperlich, Matthias Kappler, Markus Glaß, Antje Güttler, Marina Petrenko, Jonas Pyko, Tony Gutschner, Frank Tavasol, Dirk Vordermark, Matthias Bache","doi":"10.1186/s13036-025-00611-y","DOIUrl":"10.1186/s13036-025-00611-y","url":null,"abstract":"","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":"17"},"PeriodicalIF":6.5,"publicationDate":"2026-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145917680","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}
Pub Date : 2026-01-08DOI: 10.1186/s13036-025-00614-9
Changchuan Ye, Yan Zhang, Jie Zhang, Menglei Shi, Feixue Nie, Qinghua Liu
Background: Vitamin K2 (VK2), as a derivative of the menaquinone family, plays an important role in the prevention of osteoporosis and cardiovascular calcification. The realization of the industrialization of VK2 and the reduction of its production cost have become the focus of attention.
Results: In this work, an E. coli strain with high VK2 accumulation was constructed through rational metabolic engineering and stepwise improvement based on regulatory metabolic information and CRISPR/Cas9-mediated gene knockout. We first constructed a recombinant E. coli strain BW-T7/MU to produce menaquinol-8 (MKH2-8, a reduced form of VK2) by overexpressing menA and ubiE genes, which encoding the rate-limiting enzymes of the menaquinol pathway. After 24 h and 48 h of fermentation, this strain BW-T7/MU reach a titer of 303 mg/L and 232 mg/L. Secondly, we overexpressed different related genes wrbA (oxidative stress mitigation), qorB (reduction of quinones) and menF (conversion of chorismate to isochorismate), respectively. Among these recombinant strains, the strain BW-T7/MUW (overexpressing menA, ubiE and wrbA genes) reached the highest titer of VK2 after 48 h of fermentation. The optimization of the medium led to an increase in the accumulation of VK2. Subsequently, the rational metabolic engineering of gene knockout further increased the titer of VK2. The recombinant strain ΔB/MUW was selected as the dominant strain for further optimization, with a high VK2 titer of 724 mg/L. A final attempt is to overexpress ispB gene to increased flux of isoprenoid side chain synthesis, resulting in strain ΔB/MUWI with a titer of 859 mg/L in a shake flask and 1360 mg/L in a 5 L fermenter after 48 h cultivation.
Conclusions: The stepwise engineering strategy raised the VK2 titer from the initial 303 mg/L to 859 mg/L through rational pathway modification and systematic gene expression. Further optimization in batch fermentation increased the VK2 titer to 1360 mg/L, which highlights the strong engineering impact of our strategy.
{"title":"Metabolic engineering of Escherichia coli BW25113 for the production of Vitamin K<sub>2</sub> based on CRISPR/Cas9 mediated gene knockout and metabolic pathway modification.","authors":"Changchuan Ye, Yan Zhang, Jie Zhang, Menglei Shi, Feixue Nie, Qinghua Liu","doi":"10.1186/s13036-025-00614-9","DOIUrl":"10.1186/s13036-025-00614-9","url":null,"abstract":"<p><strong>Background: </strong>Vitamin K<sub>2</sub> (VK<sub>2</sub>), as a derivative of the menaquinone family, plays an important role in the prevention of osteoporosis and cardiovascular calcification. The realization of the industrialization of VK<sub>2</sub> and the reduction of its production cost have become the focus of attention.</p><p><strong>Results: </strong>In this work, an E. coli strain with high VK<sub>2</sub> accumulation was constructed through rational metabolic engineering and stepwise improvement based on regulatory metabolic information and CRISPR/Cas9-mediated gene knockout. We first constructed a recombinant E. coli strain BW-T7/MU to produce menaquinol-8 (MKH<sub>2</sub>-8, a reduced form of VK<sub>2</sub>) by overexpressing menA and ubiE genes, which encoding the rate-limiting enzymes of the menaquinol pathway. After 24 h and 48 h of fermentation, this strain BW-T7/MU reach a titer of 303 mg/L and 232 mg/L. Secondly, we overexpressed different related genes wrbA (oxidative stress mitigation), qorB (reduction of quinones) and menF (conversion of chorismate to isochorismate), respectively. Among these recombinant strains, the strain BW-T7/MUW (overexpressing menA, ubiE and wrbA genes) reached the highest titer of VK<sub>2</sub> after 48 h of fermentation. The optimization of the medium led to an increase in the accumulation of VK<sub>2</sub>. Subsequently, the rational metabolic engineering of gene knockout further increased the titer of VK<sub>2</sub>. The recombinant strain ΔB/MUW was selected as the dominant strain for further optimization, with a high VK<sub>2</sub> titer of 724 mg/L. A final attempt is to overexpress ispB gene to increased flux of isoprenoid side chain synthesis, resulting in strain ΔB/MUWI with a titer of 859 mg/L in a shake flask and 1360 mg/L in a 5 L fermenter after 48 h cultivation.</p><p><strong>Conclusions: </strong>The stepwise engineering strategy raised the VK<sub>2</sub> titer from the initial 303 mg/L to 859 mg/L through rational pathway modification and systematic gene expression. Further optimization in batch fermentation increased the VK<sub>2</sub> titer to 1360 mg/L, which highlights the strong engineering impact of our strategy.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":"29"},"PeriodicalIF":6.5,"publicationDate":"2026-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145933507","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}
Pub Date : 2026-01-08DOI: 10.1186/s13036-025-00609-6
Yao Xiao, Xuxu Li, Lijin Jiang, Yiji Zhao, Lei Wang, Yan Feng
{"title":"Construction of Escherichia coli L-isoleucine cell factories based on propionate pathway.","authors":"Yao Xiao, Xuxu Li, Lijin Jiang, Yiji Zhao, Lei Wang, Yan Feng","doi":"10.1186/s13036-025-00609-6","DOIUrl":"10.1186/s13036-025-00609-6","url":null,"abstract":"","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":"28"},"PeriodicalIF":6.5,"publicationDate":"2026-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145933500","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}
Pub Date : 2026-01-07DOI: 10.1186/s13036-025-00602-z
Bruno F Gaag, Peter Tang, Oliver Klein, Simon Moosburner, Agnes K Böhm, Theresa Lohmann, Jonas K Wieland, Victoria Contes, Yijun Zhou, Eriselda Keshi, Luna Haderer, Eric Metzler, Verena Schöwel-Wolf, Simone Spuler, Jens-Carsten Rückert, Johann Pratschke, Igor M Sauer, Marco N Andreas, Karl H Hillebrandt
{"title":"Comparison of different decellularization protocols for porcine centrum tendineum diaphragmatis and diaphragmatic muscle - a base for effective recellularization.","authors":"Bruno F Gaag, Peter Tang, Oliver Klein, Simon Moosburner, Agnes K Böhm, Theresa Lohmann, Jonas K Wieland, Victoria Contes, Yijun Zhou, Eriselda Keshi, Luna Haderer, Eric Metzler, Verena Schöwel-Wolf, Simone Spuler, Jens-Carsten Rückert, Johann Pratschke, Igor M Sauer, Marco N Andreas, Karl H Hillebrandt","doi":"10.1186/s13036-025-00602-z","DOIUrl":"10.1186/s13036-025-00602-z","url":null,"abstract":"","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":"16"},"PeriodicalIF":6.5,"publicationDate":"2026-01-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145917731","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}
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