Biotechnology and Bioengineering最新文献_第10页

Special Issue on “Recovery of Biological Products: Past, Present, Future” 生物产品回收 "特刊：过去、现在和未来 "特刊。

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-07-11 DOI: 10.1002/bit.28792

Suzanne S. Farid, Nihal Tugcu, Arne Staby

引用次数: 0

The characterization of cell traction force on nonflat surfaces with different curvature by elastic hydrogel microspheres 弹性水凝胶微球对不同曲率非平坦表面上细胞牵引力的表征。

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-07-08 DOI: 10.1002/bit.28802

Yuqing Dong, Cong Wang, Xin Ding, Xingquan Ma, Rong Huang, Moxiao Li, Qingzhen Yang

{"title":"The characterization of cell traction force on nonflat surfaces with different curvature by elastic hydrogel microspheres","authors":"Yuqing Dong, Cong Wang, Xin Ding, Xingquan Ma, Rong Huang, Moxiao Li, Qingzhen Yang","doi":"10.1002/bit.28802","DOIUrl":"10.1002/bit.28802","url":null,"abstract":"It is of great importance to study the detachment/attachment behaviors of cells (cancer cell, immune cell, and epithelial cell), as they are closely related with tumor metastasis, immunoreaction, and tissue development at variety scales. To characterize the detachment/attachment during the interaction between cells and substrate, some researchers proposed using cell traction force (CTF) as the indicator. To date, various strategies have been developed to measure the CTF. However, these methods only realize the measurements of cell passive forces on flat cases. To quantify the active CTF on nonflat surfaces, which can better mimic the in vivo case, we employed elastic hydrogel microspheres as a force sensor. The microspheres were fabricated by microfluidic chips with controllable size and mechanical properties to mimic substrate. Cells were cultured on microsphere and the CTF led to the deformation of microsphere. By detecting the morphology information, the CTF exerted by attached cells can be calculated by the in-house numerical code. Using these microspheres, the CTF of various cells (including tumor cell, immunological cell, and epithelium cell) were successfully obtained on nonflat surfaces with different curvature radii. The proposed method provides a versatile platform to measure the CTF with high precision and to understand the detachment/attachment behaviors during physiology processes.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"121 11","pages":"3537-3550"},"PeriodicalIF":3.5,"publicationDate":"2024-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141558103","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}

引用次数: 0

Revisiting the impact of genomic hot spots: C12orf35 locus as a hot spot and engineering target 重新审视基因组热点的影响：作为热点和工程目标的 C12orf35 基因座

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-07-08 DOI: 10.1002/bit.28801

Kyuhee Cho, Jae Seong Lee

{"title":"Revisiting the impact of genomic hot spots: C12orf35 locus as a hot spot and engineering target","authors":"Kyuhee Cho, Jae Seong Lee","doi":"10.1002/bit.28801","DOIUrl":"10.1002/bit.28801","url":null,"abstract":"Traditional Chinese hamster ovary (CHO) cell line development is based on random integration (RI) of transgene that causes clonal variation and subsequent large-scale clone screening. Therefore, site-specific integration (SSI) of transgenes into genomic hot spots has recently emerged as an alternative method for cell line development. However, the specific mechanisms underlying hot spot site formation remain unclear. In this study, we aimed to generate landing pad (LP) cell lines via the RI of transgenes encoding fluorescent reporter proteins flanked by recombination sites to facilitate recombinase-mediated cassette exchange. The RI-based LP cell line expressing high reporter levels with spontaneous C12orf35 locus deletion exhibited similar reporter fluorescent protein levels compared to targeted integrants with an identical reporter LP construct at the CHO genome hot spot, the C12orf35 locus. Additionally, Resf1, a C12orf35 locus gene, knockout (KO) in the RI-based LP cell line with conserved C12orf35 increased reporter expression levels, comparable to those in cell lines with C12orf35 locus disruption. These results indicate that the effect of SSI into the C12orf35 locus, a genomic hot spot, on high-level transgene expression was caused by C12orf35 disruption. In contrast to C12orf35 KO, KO at other well-known hot spot sites at specific loci of genes, including Fer1L4, Hprt1, Adgrl4, Clcc1, Dop1b, and Ddc, did not increase transgene expression. Overall, our findings suggest that C12orf35 is a promising engineering target and a hot spot for SSI-based cell line development.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"121 11","pages":"3642-3649"},"PeriodicalIF":3.5,"publicationDate":"2024-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.28801","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141558102","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Fine and combinatorial regulation of key metabolic pathway for enhanced β-alanine biosynthesis with non-inducible Escherichia coli 利用非诱导型大肠杆菌对关键代谢途径进行精细和组合调控，以增强 β-丙氨酸的生物合成。

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-07-08 DOI: 10.1002/bit.28799

Hai-Yan Zhou, Wen-Qing Ding, Xi Zhang, Hong-Yu Zhang, Zhong-Ce Hu, Zhi-Qiang Liu, Yu-Guo Zheng

{"title":"Fine and combinatorial regulation of key metabolic pathway for enhanced β-alanine biosynthesis with non-inducible Escherichia coli","authors":"Hai-Yan Zhou, Wen-Qing Ding, Xi Zhang, Hong-Yu Zhang, Zhong-Ce Hu, Zhi-Qiang Liu, Yu-Guo Zheng","doi":"10.1002/bit.28799","DOIUrl":"10.1002/bit.28799","url":null,"abstract":"β-Alanine is the only β-amino acid in nature and one of the most important three-carbon chemicals. This work was aimed to construct a non-inducible β-alanine producer with enhanced metabolic flux towards β-alanine biosynthesis in Escherichia coli. First of all, the assembled E. coli endogenous promoters and 5′-untranslated regions (PUTR) were screened to finely regulate the combinatorial expression of genes panDBS and aspBCG for an optimal flux match between two key pathways. Subsequently, additional copies of key genes (panDBSK104S and ppc) were chromosomally introduced into the host A1. On these bases, dynamical regulation of the gene thrA was performed to reduce the carbon flux directed in the competitive pathway. Finally, the β-alanine titer reached 10.25 g/L by strain A14-R15, 361.7% higher than that of the original strain. Under fed-batch fermentation in a 5-L fermentor, a titer of 57.13 g/L β-alanine was achieved at 80 h. This is the highest titer of β-alanine production ever reported using non-inducible engineered E. coli. This metabolic modification strategy for optimal carbon flux distribution developed in this work could also be used for the production of various metabolic products.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"121 10","pages":"3297-3310"},"PeriodicalIF":3.5,"publicationDate":"2024-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141558101","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}

引用次数: 0

Optical discrimination of pathological red blood cells 病理红细胞的光学鉴别

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-07-07 DOI: 10.1002/bit.28798

Michele Vergari, Benedetta Niccolini, Dario Pitocco, Alessandro Rizzi, Gabriele Ciasca, Marco de Spirito, Luca Gavioli

引用次数: 0

High-throughput G protein-coupled receptor-based autocrine screening for secondary metabolite production in yeast 基于 G 蛋白偶联受体的高通量自分泌筛选酵母中次生代谢物的产生。

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-07-07 DOI: 10.1002/bit.28797

Tatyana E. Saleski, Huadong Peng, Bettina Lengger, Jinglin Wang, Michael Krogh Jensen, Emil D. Jensen

{"title":"High-throughput G protein-coupled receptor-based autocrine screening for secondary metabolite production in yeast","authors":"Tatyana E. Saleski, Huadong Peng, Bettina Lengger, Jinglin Wang, Michael Krogh Jensen, Emil D. Jensen","doi":"10.1002/bit.28797","DOIUrl":"10.1002/bit.28797","url":null,"abstract":"Biosensors are valuable tools in accelerating the test phase of the design-build-test-learn cycle of cell factory development, as well as in bioprocess monitoring and control. G protein-coupled receptor (GPCR)-based biosensors enable cells to sense a wide array of molecules and environmental conditions in a specific manner. Due to the extracellular nature of their sensing, GPCR-based biosensors require compartmentalization of distinct genotypes when screening production levels of a strain library to ensure that detected levels originate exclusively from the strain under assessment. Here, we explore the integration of production and sensing modalities into a single Saccharomyces cerevisiae strain and compartmentalization using three different methods: (1) cultivation in microtiter plates, (2) spatial separation on agar plates, and (3) encapsulation in water-in-oil-in-water double emulsion droplets, combined with analysis and sorting via a fluorescence-activated cell sorting machine. Employing tryptamine and serotonin as proof-of-concept target molecules, we optimize biosensing conditions and demonstrate the ability of the autocrine screening method to enrich for high producers, showing the enrichment of a serotonin-producing strain over a nonproducing strain. These findings illustrate a workflow that can be adapted to screening for a wide range of complex chemistry at high throughput using commercially available microfluidic systems.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"121 10","pages":"3283-3296"},"PeriodicalIF":3.5,"publicationDate":"2024-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.28797","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141554204","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Modulating metabolism through synthetic biology: Opportunities for two-stage fermentation 通过合成生物学调节新陈代谢：两阶段发酵的机遇。

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-07-06 DOI: 10.1002/bit.28791

Yixin Rong, Sheila Ingemann Jensen, John M. Woodley, Alex Toftgaard Nielsen

引用次数: 0

An engineered ultrahigh affinity bi-paratopic uPAR targeting agent confers enhanced tumor targeting 一种超高亲和力双伞uPAR靶向制剂可增强肿瘤靶向性。

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-07-04 DOI: 10.1002/bit.28790

Gerald M. Cherf, Robert B. Lee, Nishant Mehta, Claire Clifford, Kathleen Torres, James R. Kintzing, Jennifer R. Cochran

引用次数: 0

Combinatorial metabolic engineering of Bacillus subtilis for menaquinone-7 biosynthesis 对枯草芽孢杆菌进行组合代谢工程，以合成脑醌-7。

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-07-04 DOI: 10.1002/bit.28800

Xian Sun, Xinyu Bi, Guyue Li, Shixiu Cui, Xianhao Xu, Yanfeng Liu, Jianghua Li, Guocheng Du, Xueqin Lv, Long Liu

{"title":"Combinatorial metabolic engineering of Bacillus subtilis for menaquinone-7 biosynthesis","authors":"Xian Sun, Xinyu Bi, Guyue Li, Shixiu Cui, Xianhao Xu, Yanfeng Liu, Jianghua Li, Guocheng Du, Xueqin Lv, Long Liu","doi":"10.1002/bit.28800","DOIUrl":"10.1002/bit.28800","url":null,"abstract":"Menaquinone-7 (MK-7), a form of vitamin K2, supports bone health and prevents arterial calcification. Microbial fermentation for MK-7 production has attracted widespread attention because of its low cost and short production cycles. However, insufficient substrate supply, unbalanced precursor synthesis, and low catalytic efficiency of key enzymes severely limited the efficiency of MK-7 synthesis. In this study, utilizing Bacillus subtilis BSAT01 (with an initial MK-7 titer of 231.0 mg/L) obtained in our previous study, the glycerol metabolism pathway was first enhanced to increase the 3-deoxy-arabino-heptulonate 7-phosphate (DHAP) supply, which led to an increase in MK-7 titer to 259.7 mg/L. Subsequently, a combination of knockout strategies predicted by the genome-scale metabolic model etiBsu1209 was employed to optimize the central carbon metabolism pathway, and the resulting strain showed an increase in MK-7 production from 259.7 to 318.3 mg/L. Finally, model predictions revealed the methylerythritol phosphate pathway as the major restriction pathway, and the pathway flux was increased by heterologous introduction (Introduction of Dxs derived from Escherichia coli) and fusion expression (End-to-end fusion of two enzymes by a linker peptide), resulting in a strain with a titer of 451.0 mg/L in a shake flask and 474.0 mg/L in a 50-L bioreactor. This study achieved efficient MK-7 synthesis in B. subtilis, laying the foundation for large-scale MK-7 bioproduction.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"121 10","pages":"3338-3350"},"PeriodicalIF":3.5,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141533648","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}

引用次数: 0

Non-industrial production of therapeutic lentiviral vectors: How to provide vectors to academic CAR-T 治疗性慢病毒载体的非工业化生产：如何为 CAR-T 学术研究提供载体？

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-07-04 DOI: 10.1002/bit.28794

Viktoriya Keyer, Laura Syzdykova, Bakytkali Ingirbay, Elena Sedova, Gulzat Zauatbayeva, Tolganay Kulatay, Alexandr Shevtsov, Alexandr V. Shustov

{"title":"Non-industrial production of therapeutic lentiviral vectors: How to provide vectors to academic CAR-T","authors":"Viktoriya Keyer, Laura Syzdykova, Bakytkali Ingirbay, Elena Sedova, Gulzat Zauatbayeva, Tolganay Kulatay, Alexandr Shevtsov, Alexandr V. Shustov","doi":"10.1002/bit.28794","DOIUrl":"10.1002/bit.28794","url":null,"abstract":"Bringing effective cancer therapy in the form of chimeric antigen receptor technology to untapped markets faces numerous challenges, including a global shortage of therapeutic lentiviral or retroviral vectors on which all current clinical therapies using genetically modified T cells are based. Production of these lentiviral vectors in academic settings in principle opens the way to local production of therapeutic cells, which is the only economically viable approach to make this therapy available to patients in developing countries.The conditions for obtaining and concentrating lentiviral vectors have been optimized and described. The calcium phosphate precipitation method was found to be suitable for transfecting high cell-density cultures, a prerequisite for high titers. We describe protocols for gradually increasing production from 6-well plates to P100 plates, T-175 flasks, and 5-layer stacks while maintaining high titers, >108 transducing units. Concentration experiments using ultracentrifugation revealed the advantage of lower centrifugation speeds compared to competing protocols. The resulting batches of lentiviral vectors had a titer of 1010 infectious particles and were used to transduce primary human T lymphocytes generating chimeric antigen receptor T cells, the quality of which was checked and found potential applicability for treatment.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"121 10","pages":"3252-3268"},"PeriodicalIF":3.5,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141497113","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}

引用次数: 0