Journal of biotechnology最新文献

Establishment of the REMBAC-cassette, a rapid, efficient and manifold BacMam tool for recombinant protein expression.

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-01-03 DOI: 10.1016/j.jbiotec.2024.12.011

Manuel Reithofer, Sophie Huber, Reingard Grabherr

{"title":"Establishment of the REMBAC-cassette, a rapid, efficient and manifold BacMam tool for recombinant protein expression.","authors":"Manuel Reithofer, Sophie Huber, Reingard Grabherr","doi":"10.1016/j.jbiotec.2024.12.011","DOIUrl":"10.1016/j.jbiotec.2024.12.011","url":null,"abstract":"Efficient recombinant protein production requires mammalian stable cell lines or often relies on inefficient transfection processes. Baculoviral transduction of mammalian cells (BacMam) offers cost-effective and robust gene transfer and straightforward scalability. The advantages over conventional approaches are, no need of high biosafety level laboratories, efficient transduction of various cell types and transfer of large transgenes into host cells. In our study, we aim to develop a high expression cassette to increase yields of baculoviral transduction. The establishment follows a sequential approach by first identifying the strongest promoter, followed by intron and WPRE sequences as enhancer elements for transcription and translation. The resulting REMBAC-cassette was compared to conventional transfection in suspension and adherent cells. Irrespective of the cell line, transduction reached nearly 100 % efficiency and led to almost 10-fold increases of gene expression levels. We confirmed these results in larger scale with batch and fed-batch cultivations. Finally, expression of different soluble proteins with high degrees of complexity confirmed the versatility of our established cassette. Overall, the REMBAC-cassette incorporated into the BacMam platform is a manifold tool offering advantages over standard transfection, in the scalability, efficiency and gene expression, which results in higher yields, shorter cultivation times and consequently cost-effective production processes.","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":" ","pages":"183-192"},"PeriodicalIF":4.1,"publicationDate":"2025-01-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142927174","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

Improved biosynthesis of tyrosol by epigenetic modification-based regulation and metabolic engineering in Saccharomyces cerevisiae.

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-12-31 DOI: 10.1016/j.jbiotec.2024.12.013

Guoli Wang, Xiqin Liang, Zhenke Wu, Bengui Fan, Jun Wang, Qiusheng Zheng, Defang Li, Tianyue An

{"title":"Improved biosynthesis of tyrosol by epigenetic modification-based regulation and metabolic engineering in Saccharomyces cerevisiae.","authors":"Guoli Wang, Xiqin Liang, Zhenke Wu, Bengui Fan, Jun Wang, Qiusheng Zheng, Defang Li, Tianyue An","doi":"10.1016/j.jbiotec.2024.12.013","DOIUrl":"10.1016/j.jbiotec.2024.12.013","url":null,"abstract":"Aromatic amino acids and their derivatives are high value chemicals widely used in food, pharmaceutical and feed industries. Current preparation methods for aromatic amino acid products are fraught with limitations. In this study, the efficient biosynthesis of aromatic amino acid compound tyrosol was investigated by epigenetic modification-based regulation and optimization of the biosynthetic pathway of aromatic amino acids. The production of tyrosol was significantly improved by the overexpression of m6A modification writer Ime4 and reader Pho92, and the positive regulator Gcr2. Introduction of Bbxfpk and deletion of Gpp1 further improved tyrosol production. Then the feedback inhibition of the shikimate pathway was relieved by the mutants Aro4K229L and Aro7G141S. The final tyrosol producing engineered strain was constructed by the deletion of PHA2, replacement of the native promoter of ARO10 with the strong promoter PGK1p, and introduction of tyrosine decarboxylase PcAAS. In the background of m6A modification regulation, this strain ultimately produced 954.69 ± 43.72 mg/L of tyrosol, promoted by 61.7-fold in shake-flask fermentation.","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":" ","pages":"175-182"},"PeriodicalIF":4.1,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142921807","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

Converting multiple hydrophobic aromatic plastic monomers into a single water-soluble substrate to increase bioavailability for the synthesis of polyhydroxyalkanoates by bacteria using batch, fed batch and continuous cultivation.

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-12-26 DOI: 10.1016/j.jbiotec.2024.12.008

Karthika Balusamy, Balaji V Rokade, Manuel Bruch, Meg Walsh, William Casey, Federico Cerrone, Bryan Dalton, Patrick Jerome Guiry, Tanja Narančić, Kevin E O'Connor

{"title":"Converting multiple hydrophobic aromatic plastic monomers into a single water-soluble substrate to increase bioavailability for the synthesis of polyhydroxyalkanoates by bacteria using batch, fed batch and continuous cultivation.","authors":"Karthika Balusamy, Balaji V Rokade, Manuel Bruch, Meg Walsh, William Casey, Federico Cerrone, Bryan Dalton, Patrick Jerome Guiry, Tanja Narančić, Kevin E O'Connor","doi":"10.1016/j.jbiotec.2024.12.008","DOIUrl":"https://doi.org/10.1016/j.jbiotec.2024.12.008","url":null,"abstract":"We demonstrate the proof of concept of increasing the bioavailability of carbon substrates, derived from plastic waste, for their conversion to the biodegradable polymer polyhydroxyalkanoate [PHA] by bacteria and test various approaches to PHA accumulation through batch, fed batch and continuous culture. Styrene, ethylbenzene, and toluene are produced from the pyrolysis of mixed plastic waste (Kaminsky, 2021; Miandad et al., 2017), but they are volatile and poorly soluble in water making them difficult to work with in aqueous fermentation systems. By chemically converting these aromatic compounds to benzoic acid, and subsequently to its sodium salt, we increased the solubility and reduced the volatility of the substrate supplied to Pseudomonas putida CA-3 to accumulate polyhydroxyalkanoates. 1L scale batch, fed batch, and continuous fermentations were carried out; the fed batch fermentation resulted in the maximum volumetric PHA productivity of 61.67 ± 7.34mgL-1 h-1; while batch and continuous, at a dilution rate, d = 0.2h-1, fermentations resulted in 13.30 ± 0.01 and 4.06 ± 0.01mgL-1 h-1 of PHA respectively.","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":" ","pages":""},"PeriodicalIF":4.1,"publicationDate":"2024-12-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142894667","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

The role of biosilica and its potential for sensing technologies: A review.

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-12-25 DOI: 10.1016/j.jbiotec.2024.12.010

Roberta Anjos de Jesus, Ivani Meneses Costa, Katlin Ivon Barrios Eguiluz, Giancarlo Richard Salazar-Banda

引用次数: 0

Alleviation of cadmium toxicity and minimizing its accumulation in rice plants by methyl jasmonate: performance and mechanisms.

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-12-24 DOI: 10.1016/j.jbiotec.2024.12.009

Ting Wei, Hong Li, Yuyao Wang, Ming Chi, Junkang Guo, Honglei Jia, Chao Zhang

{"title":"Alleviation of cadmium toxicity and minimizing its accumulation in rice plants by methyl jasmonate: performance and mechanisms.","authors":"Ting Wei, Hong Li, Yuyao Wang, Ming Chi, Junkang Guo, Honglei Jia, Chao Zhang","doi":"10.1016/j.jbiotec.2024.12.009","DOIUrl":"https://doi.org/10.1016/j.jbiotec.2024.12.009","url":null,"abstract":"Heavy metal pollution is a worldwide problem that threaten agricultural production and human health. Methyl jasmonate (MeJA) is a phytohormone that could enhance plant resistance against various stresses. However, the mechanism of MeJA in cadmium (Cd) uptake, distribution, and translocation in rice plants remains elusive. In this study, we found that the Cd induced-growth inhibition was ameliorated by MeJA. Upon MeJA application, Cd content in root and shoot was decreased by 10.15% and 36.39%, which paralleled with less Cd2+ influx of rice roots and depressed expression of the cation transporters (OsNramp1 and OsNramp5). The subcellular distribution revealed that MeJA enriched Cd distribution in cell wall, which was accompanied by increased cell wall thickness and altered cell wall polysaccharide (pectin, cellulose, hemicellulose) content, meanwhile, the Cd content in pectin, cellulose, hemicellulose was increased, the FTIR analysis implied that functional groups (especially -OH and COO-) on cell wall were involved in Cd fixation. The root to shoot translocation of Cd was hindered by exogenous MeJA, this was validated by the decreased expression of OsHMA2 in root and declined Cd level in xylem sap. Overall, our results revealed that MeJA could act as a foliar resistance control substance to reduce Cd accumulation in rice plants. The detailed molecular mechanisms of MeJA in Cd detoxification in plants still need further investigation.","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":" ","pages":""},"PeriodicalIF":4.1,"publicationDate":"2024-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142894664","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

Molecular mechanism of GSH metabolism and autophagy in NAC-promoted recombinant human serum albumin and follicle stimulating hormone beta fusion protein secretion in Pichia pastoris.

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-12-20 DOI: 10.1016/j.jbiotec.2024.12.006

Gang Luo, Wen Yang, Zijian Geng, Yiyi Cheng, Yingqing Xu, Yimeng Xiao, Jiying Liu

{"title":"Molecular mechanism of GSH metabolism and autophagy in NAC-promoted recombinant human serum albumin and follicle stimulating hormone beta fusion protein secretion in Pichia pastoris.","authors":"Gang Luo, Wen Yang, Zijian Geng, Yiyi Cheng, Yingqing Xu, Yimeng Xiao, Jiying Liu","doi":"10.1016/j.jbiotec.2024.12.006","DOIUrl":"10.1016/j.jbiotec.2024.12.006","url":null,"abstract":"The Pichia pastoris expression system is a favorable platform for production of pharmaceutical proteins. Treatment of strains with N-acetyl-L-cysteine (NAC) has been shown to enhance the yield of recombinant proteins, thereby contributing to a reduction in production costs. However, the specific mechanism of action of NAC remains unclear. Previous research has indicated that glutathione (GSH) and autophagy are involved in the increased production of human serum albumin and porcine follicle-stimulating hormone β (HSA-pFSHβ) by NAC. This study investigated the potential interaction between GSH and autophagy in the production of HSA-pFSHβ. The findings indicated that sulfhydryl-free antioxidants such as melatonin, vitamin C, or vitamin E did not exhibit similar effects to NAC in enhancing HSA-pFSHβ yield. Moreover, NAC was found to enhance HSA-pFSHβ production by modulating GSH metabolism to reduce GSH consumption, increase total GSH levels, as well as glutathione peroxidase (GSH-Px) and glutathione reductase (GR) activities. Additionally, inhibition of autophagy through disruption of autophagy scaffolding proteins Atg1 or Atg11 led to an increase in recombinant HSA-pFSHβ production. Furthermore, NAC significantly decreased the phosphorylation of Slt2, and the absence of the SLT2 gene influenced the effect of NAC on HSA-pFSHβ secretion by modulating mitophagy and GSH metabolism. In conclusion, these results suggest a complex interplay between GSH metabolism and autophagy in the regulation of NAC-induced HSA-pFSHβ secretion.","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":" ","pages":"146-157"},"PeriodicalIF":4.1,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142877145","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

Potency by design: Novel insights in transfection and purification for manufacturing of rAAV gene therapy vectors.

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-12-17 DOI: 10.1016/j.jbiotec.2024.12.007

Andreas Maccani, Robert Pachlinger, Peter Eisenhut, Sabine Unterthurner, Susanne Heider, Christoph Repik, Stefan Reuberger, Peter Andorfer, Johannes Lengler, Bernhard Kinastberger, Dieter Seczer, Petra Gruber, Alexander Apschner, Lucia Micutkova

{"title":"Potency by design: Novel insights in transfection and purification for manufacturing of rAAV gene therapy vectors.","authors":"Andreas Maccani, Robert Pachlinger, Peter Eisenhut, Sabine Unterthurner, Susanne Heider, Christoph Repik, Stefan Reuberger, Peter Andorfer, Johannes Lengler, Bernhard Kinastberger, Dieter Seczer, Petra Gruber, Alexander Apschner, Lucia Micutkova","doi":"10.1016/j.jbiotec.2024.12.007","DOIUrl":"10.1016/j.jbiotec.2024.12.007","url":null,"abstract":"This study investigates the crucial role of transfection methods in the manufacturability and potency of recombinant adeno-associated virus (rAAV) gene therapies. By employing a novel analytical approach, multiplex digital PCR (dPCR), we evaluated the impact of different transfection reagents and conditions on the scalability and quality of rAAV. Our research demonstrates that the selection of transfection approach significantly influences not only the yield and ease of scale-up but also the potency of the final product. Importantly, later changes to transfection parameters established in the early stages of development can be challenging, potentially compromising product quality and leading to comparability issues. Leveraging multiplex dPCR has proven instrumental in guiding these early-stage decisions, ensuring a reliable manufacturing process that consistently delivers high-quality therapeutic products. Our findings highlight the importance of optimizing transfection strategies early in development to guarantee the successful production of potent and cost-effective gene therapies, ultimately impacting patient accessibility.","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":" ","pages":"127-132"},"PeriodicalIF":4.1,"publicationDate":"2024-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142864376","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

Bioinformatics-assisted mining and design of novel pullulanase suitable for starch cold hydrolysis.

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-12-15 DOI: 10.1016/j.jbiotec.2024.12.005

Xin Wang, Zixing Wang, Xueting Zhang, Yanli Zhang, Wenxia Zhang, Yu Zhang, Xuecheng Zhang, Yazhong Xiao, Yinliang Zhang, Wei Fang

{"title":"Bioinformatics-assisted mining and design of novel pullulanase suitable for starch cold hydrolysis.","authors":"Xin Wang, Zixing Wang, Xueting Zhang, Yanli Zhang, Wenxia Zhang, Yu Zhang, Xuecheng Zhang, Yazhong Xiao, Yinliang Zhang, Wei Fang","doi":"10.1016/j.jbiotec.2024.12.005","DOIUrl":"10.1016/j.jbiotec.2024.12.005","url":null,"abstract":"Cold-active pullulanases with good catalytic performance possess promising applications in cold hydrolysis of starch. Adopting bioinformatics-assisted mining strategies, 7 candidate cold-active pullulanases were initially screened out from IMG/MER database. Among the candidates, PulBs exhibited good thermostability and the highest specific activity of 147.4 U/mg. The half-life of PulBs was about 200 h at 35 °C. Employing PulBs as the initial enzyme, the active-site design of FuncLib was implemented to enhance the activity. The design PulBs-20 exhibited an enhanced specific activity of 209.9 U/mg, which was 1.4 times that of PulBs. Furthermore, the thermostability of PulBs-20 was augmented, with a half-life of 250 h at 35 °C. When applied in the cold hydrolysis of starch, PulBs-20 can effectively enhance the hydrolysis effect of raw starch. Supplemented with the raw starch-hydrolyzing α-amylase AmyZ1 and PulBs-20, the hydrolysis rate of raw corn starch increased to 53.5 %, which was 1.3 times that of using AmyZ1 alone. Due to its high hydrolysis activity and good thermostability, PulBs-20 can serve as an efficient accessory enzyme in starch cold hydrolysis.","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":" ","pages":"106-116"},"PeriodicalIF":4.1,"publicationDate":"2024-12-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142835780","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

The physiological role of gamma-aminobutyric acid in relieving the effect of furfural inhibitor for improvement the production of lipid in D. intermedius Z₈.

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-12-15 DOI: 10.1016/j.jbiotec.2024.12.004

Min Li, Zhi Li, Lei Wei, Ge Bai, Saijin Wei, Linbin Zhou, Hanguang Li

{"title":"The physiological role of gamma-aminobutyric acid in relieving the effect of furfural inhibitor for improvement the production of lipid in D. intermedius Z8.","authors":"Min Li, Zhi Li, Lei Wei, Ge Bai, Saijin Wei, Linbin Zhou, Hanguang Li","doi":"10.1016/j.jbiotec.2024.12.004","DOIUrl":"10.1016/j.jbiotec.2024.12.004","url":null,"abstract":"In order to improve the production and quality of lipid of D. intermedius Z8 under the furfural stress environment, different exogenous γ-aminobutyric acid (GABA) addition strategies and their reasons for improving the fermentation performance were investigated. For this purpose, the effect of different concentrations of furfural on the production of biomass and lipid in D. intermedius Z8 was researched. The result shows that the growth of D. intermedius Z8 nearly stopped when 1.0 g/L furfural was adopted. However, when 1.5 mmol/L GABA was used, the highest biomass and lipid production of 4.56 g/L and 1.83 g/L were obtained, respectively, which were 36.53 % and 61.95 % higher compared to the control group (the normal development BG11 medium). Additionally, the changes in microalgal cell morphology were analyzed using the scanning electron microscope (SEM) technology, and the results suggest that GABA addition could significantly mitigate the toxic effects of excessive furfural by maintaining the integrity of the cell. Simultaneously, the activities of key enzymes involved in fatty acid synthesis, such as nitrate reductase (NR), phosphatidic acid phosphatase (PAP), acetyl CoA carboxylase (ACC), and fatty acid synthase (FAS), were enhanced under the optimal condition, and thus improving lipid production. According to the results in this study, the exogenous GABA addition strategy is a simple and effective approach to improve microbial tolerance and enhance its fermentation performance.","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":" ","pages":"117-126"},"PeriodicalIF":4.1,"publicationDate":"2024-12-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142846650","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

Corrigendum to "Cell-free reduction of carboxylic acids with secreted carboxylic acid reductase" [J. Biotechnol. 382 (2024) 44-50].

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-12-10 DOI: 10.1016/j.jbiotec.2024.12.002

Dominic Goj, Stella Ebner, Melissa Horvat, Simon Arhar, Ludmila Martínková, Margit Winkler

引用次数: 0