Journal of biotechnology最新文献_第2页

Engineering an Escherichia coli surface display platform based on an autotransporter from Stenotrophomonas maltophilia: Autodisplay of enzymes with low to high molecular weight.

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-12-09 DOI: 10.1016/j.jbiotec.2024.12.003

Kunpeng Gao, Kexin Yu, Jianan Sun, Xiangzhao Mao, Hao Dong

{"title":"Engineering an Escherichia coli surface display platform based on an autotransporter from Stenotrophomonas maltophilia: Autodisplay of enzymes with low to high molecular weight.","authors":"Kunpeng Gao, Kexin Yu, Jianan Sun, Xiangzhao Mao, Hao Dong","doi":"10.1016/j.jbiotec.2024.12.003","DOIUrl":"https://doi.org/10.1016/j.jbiotec.2024.12.003","url":null,"abstract":"Surface display technology has garnered significant attention for preparing efficient whole cell catalysts, while reported carrier proteins still cannot meet the demand to display various passenger domains, especially for those with high molecular weight. This study demonstrates that the autotransporter of esterase Est7 (E7AT) from Stenotrophomonas maltophilia played a decisive role in its efficient surface display. Guided by the original signal peptide, the surface display ratio of Est7 was determined as 89.67% with the total enzymatic activity of 16.67U/mL, which was much higher than 4.60U/mL and 5.70U/mL for the signal peptides derived from pectolase B (pelB) and cholera toxin B (ctxB), respectively. Then, the E7AT unit was successfully developed to surface display proteins with varying molecular weight from 19.3kDa to 117.9kDa, showing a more effective autodisplay ability than adhesin involved in diffuse adherence (AIDA-I), ice nucleation proteins (InaK and InaP), and outer membrane proteins (lipoprotein/ompA, MltA-interacting protein A, and yiaT) systems. Additionally, a galactosidase (GAL) displayed by E7AT was employed to hydrolyze lactose, achieving a promising hydrolysis rate of 31.63% in 2h. The displayed GAL retained 63.24% and 41.41% activity in third and sixth batch, respectively, indicating the considerable potential of E7AT in developing efficient whole cell catalysts.","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":" ","pages":""},"PeriodicalIF":4.1,"publicationDate":"2024-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142813280","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

Efficient production of spermidine from Bacillus amyloliquefaciens by enhancing synthesis pathway, blocking degradation pathway and increasing precursor supply.

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-12-06 DOI: 10.1016/j.jbiotec.2024.12.001

Ziyue Zhao, Ailing Guo, Dian Zou, Zhou Li, Xuetuan Wei

{"title":"Efficient production of spermidine from Bacillus amyloliquefaciens by enhancing synthesis pathway, blocking degradation pathway and increasing precursor supply.","authors":"Ziyue Zhao, Ailing Guo, Dian Zou, Zhou Li, Xuetuan Wei","doi":"10.1016/j.jbiotec.2024.12.001","DOIUrl":"10.1016/j.jbiotec.2024.12.001","url":null,"abstract":"Spermidine has broad application potential in food, medicine and other fields. In this study, a novel Bacillus amyloliquefaciens cell factory was constructed for production of spermidine from renewablebiomass resources. Firstly, the speB gene was found to be optimal for synthesis of spermidine, and the function of SpeB was explained by amino acid sequence analysis and molecular docking. By replacing the native promoter of the speEB operon with the P43, the synthesis of spermidine was significantly enhanced in B. amyloliquefaciens HSPM1-P43speEB. After knockout of the genes yobN and bltD associated with spermidine degradation, the spermidine titer of the strain HSPM2 was further improved to 115.96 mg/L, increased by 108 % compared to HSPM1-P43speEB. Subsequently, the titer of spermidine was further increased to 277.47 mg/L through enhancing the supply of the precursor methionine by overexpression of speD. Finally, the renewable biomass resources, xylose and feather meal were optimized to produce spermidine, and the maximum titer is up to 588.10 mg/L after optimization. In conclusion, an efficient spermidine producing B. amyloliquefaciens was constructed through combinatorial metabolic engineering strategies, and the sustainable production of spermidine was achieved using the biomass resources of xylose and feather meal.","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":" ","pages":"87-96"},"PeriodicalIF":4.1,"publicationDate":"2024-12-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142794715","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 'Enhanced activation of signaling pathway by recombinant human adiponectin from genome-edited chickens' [J. Biotechnol., Vol. 395 (2024) 95-99/0168-1656].

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-12-04 DOI: 10.1016/j.jbiotec.2024.11.019

Eunhui Yoo, Hee Jung Choi, Jae Yong Han

引用次数: 0

Efficient production of RNA in Saccharomyces cerevisiae through inducing high level transcription of functional ncRNA-SRG1.

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-12-03 DOI: 10.1016/j.jbiotec.2024.11.021

Can Guo, Zhiqiang Bin, Pengjie Zhang, Jing Tang, Lianqing Wang, Yefu Chen, Dongguang Xiao, Xuewu Guo

{"title":"Efficient production of RNA in Saccharomyces cerevisiae through inducing high level transcription of functional ncRNA-SRG1.","authors":"Can Guo, Zhiqiang Bin, Pengjie Zhang, Jing Tang, Lianqing Wang, Yefu Chen, Dongguang Xiao, Xuewu Guo","doi":"10.1016/j.jbiotec.2024.11.021","DOIUrl":"10.1016/j.jbiotec.2024.11.021","url":null,"abstract":"RNA (Ribonucleic Acid) is an essential component of organisms and is widely used in the food and pharmaceutical industries. Saccharomyces cerevisiae, recognized as a safe strain, is widely used for RNA production. In this study, the S. cerevisiae W303-1a was used as a starting strain and molecular modifications were made to the functional ncRNA-SRG1 to evaluate the effect on RNA production. At the same time, its transcriptionally associated helper genes (Spt2, Spt6 and Cha4) were overexpressed and the culture medium was supplemented with serine to induce SRG1 transcription, to increase SRG1 transcription levels and investigate its effect on intracellular RNA levels. The results showed that the intracellular RNA content of the recombinant strain W303-1a-SRG1 was 10.27 %, an increase of 11.15 % compared to the starting strain (W303-1a, with an intracellular RNA content of 9.24 %). On this basis, a gene co-overexpression strain-W303-1a-SRG1-Spt6 was constructed. Simultaneously, the addition of 2 % serine strategy was used to increase the transcription level of SRG1 and RNA content of the recombinant strain. The intracellular RNA of the recombinant strain reached 11.41 %, an increase of 23.38 % compared to the starting strain (W303-1a, without serine supplementation). In addition, the growth performance of the strain was assessed by measuring the SRG1 transcription level in the strain and plotting the growth curve. Therefore, we found that improving the transcription level of ncRNA can be used as a new idea to construct S. cerevisiae with high RNA content, which provides a strong help for subsequent research in related fields. This work provides a new strategy for increasing the nucleic acid content of S. cerevisiae.","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":" ","pages":"66-75"},"PeriodicalIF":4.1,"publicationDate":"2024-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142785725","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

Development of an advanced acetaldehyde detection solution based on yeast and bacterial surface display technology.

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-11-30 DOI: 10.1016/j.jbiotec.2024.11.020

Weigeng Liu, Jiamin Cao, Di Wu, Yue Wu, Yi Qin, Yanlin Liu, Xixi Zhao, Yuyang Song

{"title":"Development of an advanced acetaldehyde detection solution based on yeast and bacterial surface display technology.","authors":"Weigeng Liu, Jiamin Cao, Di Wu, Yue Wu, Yi Qin, Yanlin Liu, Xixi Zhao, Yuyang Song","doi":"10.1016/j.jbiotec.2024.11.020","DOIUrl":"10.1016/j.jbiotec.2024.11.020","url":null,"abstract":"Acetaldehyde, a carcinogen widely present in various beverages and the natural environment, necessitates convenient and efficient detection methods. In this work, two different host strains were used to develop a sensitive, convenient, and efficient whole-cell optical biosensor for acetaldehyde detection. Acetaldehyde dehydrogenase (AldH) was displayed on the cell surface of Saccharomyces cerevisiae and E. coli using flocculin protein and the N-terminal ice nucleation protein (INP), respectively. The successful construction of yeast and bacteria surface display platforms was confirmed by laser scanning confocal microscopy. Then, the optimal AldH-display system for yeast and bacteria was confirmed. The optimum reaction conditions were determined by changing testing temperatures and pH values. The differences between the two display systems were compared. The highest whole-cell activities of yeast and bacteria under optimal conditions were 3.68 ± 0.07 U/mL/OD600 for BY-S6G and 6.95 ± 0.04 U/mL/OD600 for E-32-IrA. The strains with the best performance were chosen for the detection of acetaldehyde in wine and other beverage samples and showed substrate specificity and accuracy, in which the recovery rate ranged between 94.4 % and 110.1 %. The results demonstrated that the AldH surface display strains could be used as an optical biosensor to detect acetaldehyde in beverages and red wine.","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":" ","pages":"42-50"},"PeriodicalIF":4.1,"publicationDate":"2024-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142769262","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

Thermal degradation kinetics and purification of C-phycocyanin from thermophilic and mesophilic cyanobacteria.

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-11-29 DOI: 10.1016/j.jbiotec.2024.11.018

Supenya Chittapun, Kattiya Suwanmanee, Chatchol Kongsinkaew, Soisuda Pornpukdeewattana, Yusuf Chisti, Theppanya Charoenrat

{"title":"Thermal degradation kinetics and purification of C-phycocyanin from thermophilic and mesophilic cyanobacteria.","authors":"Supenya Chittapun, Kattiya Suwanmanee, Chatchol Kongsinkaew, Soisuda Pornpukdeewattana, Yusuf Chisti, Theppanya Charoenrat","doi":"10.1016/j.jbiotec.2024.11.018","DOIUrl":"10.1016/j.jbiotec.2024.11.018","url":null,"abstract":"The natural blue colorant C-phycocyanin (C-PC) has many potential applications but its poor heat stability limits its commercial use. This study compares the production and thermal stability of C-PC from two cyanobacteria: the thermophilic Thermosynechococcus sp. TUBT-T01 and the mesophilic Synechococcus cedrorum TISTR8589. Thermosynechococcus sp. produced nearly 1.9-fold more C-PC than S. cedrorum. Batch adsorption using a chromatographic cationic ion exchange resin (Streamline Direct HST1) was used to effectively purify the C-PC. The equilibrium adsorption capacity (Qeq) of the resin for C-PC was the highest at pH 5. At this pH, the Qeq for the thermophilic C-PC was 5.5 ± 0.1 mg mL⁻¹ , whereas for the mesophilic C-PC it was 1.5 ± 0.2 mg mL⁻¹ . Purification increased the concentration of the thermophilic C-PC by 5.9-fold, and that of mesophilic C-PC by 4.2-fold. The purity ratios of the final products from the two cyanobacteria were similar at ∼2.2. At 60 °C and pH 7, the C-PC of Thermosynechococcus sp. had ∼12-times longer half-life than the mesophilic C-PC; however, the productivity of the thermophilic C-PC was comparatively low because of a low biomass productivity of Thermosynechococcus sp.","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":" ","pages":"76-86"},"PeriodicalIF":4.1,"publicationDate":"2024-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142769264","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

A review towards sustainable analyte detection: Biomimetic inspiration in biosensor technology.

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-11-28 DOI: 10.1016/j.jbiotec.2024.11.015

Pratistha Bhagat, Lata Sheo Bachan Upadhyay

{"title":"A review towards sustainable analyte detection: Biomimetic inspiration in biosensor technology.","authors":"Pratistha Bhagat, Lata Sheo Bachan Upadhyay","doi":"10.1016/j.jbiotec.2024.11.015","DOIUrl":"10.1016/j.jbiotec.2024.11.015","url":null,"abstract":"The branch of biomimetics has witnessed a profound impact on the field of biosensor technology, reflected in sustainable analyte detection. A vast array of biosensor platforms with improved/upgraded performance have been developed and reported. No wonder the motivation from the field of biomimetics has a huge impact on generating detection systems with escalated degrees of manipulation and tunability at different levels. More recently, biomimetic biosensor technology has found potential in constructing bio-inspired materials such as aptamers, MIPs, nanozymes, DNAzymes, Synzymes, etc. to be integrated with biosensor fabrication. The establishment of a sensing setup is not limited to the bioreceptor fabrication; the construction of transducing element using biomimetic material have been reported too. Moreover, to serve a biosensing of target analyte from a fatal diseased sample different biomimetic architectures can be designed that mimic in-vivo microenvironmental surroundings to get an exact microenvironment equivalent to natural conditions leading towards designing of a precise treatment strategy. This research area is ever-evolving as there is a scope for upgradation and refinement due to advancing technologies including nanotechnology, biomimetic nanomaterials, microfluidics, optical sensors, etc. This review is an attempt to comprehend and juxtapose the very primary innovations in the field of biomimetic biosensor technology to realize its comprehensive and wide-range scope and possibilities.","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":" ","pages":"51-65"},"PeriodicalIF":4.1,"publicationDate":"2024-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142769258","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

Assessing the role of deep eutectic solvents in Yarrowia lipolytica inhibition

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-11-28 DOI: 10.1016/j.jbiotec.2024.11.016

Filipe S. Buarque , Bernardo D. Ribeiro , Mara G. Freire , Maria A.Z. Coelho , Matheus M. Pereira

{"title":"Assessing the role of deep eutectic solvents in Yarrowia lipolytica inhibition","authors":"Filipe S. Buarque , Bernardo D. Ribeiro , Mara G. Freire , Maria A.Z. Coelho , Matheus M. Pereira","doi":"10.1016/j.jbiotec.2024.11.016","DOIUrl":"10.1016/j.jbiotec.2024.11.016","url":null,"abstract":"<div><div>Yarrowia lipolytica has gained recognition as a microorganism with biological relevance and extensive biotechnological applications. Some of its features include a high enzyme secretion capacity and a high cell-density fermentation mode. Hexokinase (YlHxk) is a vital enzyme in Y. lipolytica growth since it catalyzes glucose metabolism through phosphorylation in the glycolytic pathway. Given the potential application of deep eutectic solvents (DES) as novel solvents in biotechnological processes, this study evaluated the influence of eighteen DES on the growth of Y. lipolytica. Furthermore, this work examined the effects of individual ions on the YlHxk enzyme by analyzing its enzymatic tunnel structure, molecule transport, and molecular docking. The results revealed a significant reduction in yeast growth in the presence of most DES compared to the control (medium without DES), with the exception of the [N8881]Cl: hexanoic acid (1:1) DES. The growth varied between 11.95 ± 0.60 and 0.68 ± 0.17 g dry cell weight L−1. According to the enzymatic tunnel analysis, DES components associated with the lowest microbial growth values were transported through tunnel 1. On the other hand, DES components had their pathway facilitated through tunnel 2 ([N8881]+ and hexanoic acid) and showed growth values close to the control. Molecular docking analysis identified a similarity between all the ligands in this tunnel (including substrate and product), presenting binding interactions with the ASN273 amino acid of the YlHxk active site. Combining experimental results with computational tools provided promising insights at the molecular level, while also potentially reducing analysis costs and time, paving the way for similar approaches in broad biocatalytic reactions.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"398 ","pages":"Pages 1-10"},"PeriodicalIF":4.1,"publicationDate":"2024-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142748017","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

Bone marrow-derived dendritic cells play a role in attenuating inflammation on Bothrops jararacussu venom muscle damage.

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-11-28 DOI: 10.1016/j.jbiotec.2024.11.014

N M Nery, A A Ferreira E Ferreira, H M Santana, S N Serrath, V P Reis, M V Paloschi, M D S Silva, J G S Magalhães, L F Cruz, T Y Shibayama, S S Setubal, J P Zuliani

{"title":"Bone marrow-derived dendritic cells play a role in attenuating inflammation on Bothrops jararacussu venom muscle damage.","authors":"N M Nery, A A Ferreira E Ferreira, H M Santana, S N Serrath, V P Reis, M V Paloschi, M D S Silva, J G S Magalhães, L F Cruz, T Y Shibayama, S S Setubal, J P Zuliani","doi":"10.1016/j.jbiotec.2024.11.014","DOIUrl":"10.1016/j.jbiotec.2024.11.014","url":null,"abstract":"The immune system is regulated by dendritic cells (DCs), which are highly specialized cells for presenting antigens. They are thought of as natural sentinels that start the immune response triggered by naive T cells against invasive infections. DCs participate in the initial stage of muscle damage in conjunction with monocytes, macrophages, and myogenic cells. The goal of this study was to determine whether DCs might mitigate tissue damage and aid in the regeneration of the gastrocnemius muscle following envenomation with Bothrops jararacussu venom (BjV). Mature bone marrow dendritic cells (BMDCs) were used to treat mice in an experimental envenomation model with BjV by activation with lipopolysaccharide (LPS). BMDCs were injected into the gastrocnemius muscle at the same site of the BjV injury, in a single dose, 3 h after envenomation, and envenoming effects were observed at different periods for 7 days. In both untreated (NT) and treated (T) groups tissue necrosis, leukocyte influx, and hemorrhage at the injury site were observed. Results showed an increase in serum and tissue CK as well as IL-6, TNF-α, and IL-1β release in the first hours after envenoming. In contrast, after treatment with BMDCs results obtained demonstrated an attenuated local effect with a small leukocyte influx, decreased or non-existent necrosis and hemorrhage, as well as a reduction in both serum and tissue CK levels as well as cytokine release and, consequently, the onset of a moderate regenerative process. The present study's findings concluded that BjV causes a severe inflammatory reaction at the site of injury and that treating envenoming with BMDCs in the muscle was crucial for minimizing damage to the muscle and the inflammatory reaction and promoting the early onset of the tissue repair process.","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":" ","pages":"29-40"},"PeriodicalIF":4.1,"publicationDate":"2024-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142769260","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

Creation of catalytic activity-improved hyperthermophilic PQQ-dependent aldose sugar dehydrogenase and its efficient use for high performance electro-device

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-11-28 DOI: 10.1016/j.jbiotec.2024.11.017

Miku Maeno , Yusuke Miki , Kazuki Ito , Haruhiko Sakuraba , Toshihisa Ohshima , Shin-ichiro Suye , Takenori Satomura

{"title":"Creation of catalytic activity-improved hyperthermophilic PQQ-dependent aldose sugar dehydrogenase and its efficient use for high performance electro-device","authors":"Miku Maeno , Yusuke Miki , Kazuki Ito , Haruhiko Sakuraba , Toshihisa Ohshima , Shin-ichiro Suye , Takenori Satomura","doi":"10.1016/j.jbiotec.2024.11.017","DOIUrl":"10.1016/j.jbiotec.2024.11.017","url":null,"abstract":"<div><div>PQQ-dependent aldose sugar dehydrogenase (PQQ-ASD) from the hyperthermophilic archaeon Pyrobaculum aerophilum (PaeASD) has great potential as an element for durable bioelectrodevices owing to its exceptional stability against high temperatures and across a broad pH spectrum. However, its application is constrained by low electric current output of the enzyme-immobilized electrodes, which is attributable to its low catalytic activity. A directed evolutionary approach was performed on PaeASD to improve enzyme activity, resulting in the identification of a PaeASD s24 mutant containing six amino acid substitutions, which exhibited a 16-fold higher specific activity than that of wild type. Although each single amino acid mutant among these substitutions exhibited lower enzyme activity than PaeASD s24, the double mutant R64Q/D350N showed enzyme activity comparable to that of PaeASD s24. These amino acids located in the vicinity of coenzyme PQQ within the PaeASD molecule are also highly conserved with those of PQQ-ASDs reported to date. Thus, these amino acids play crucial roles in the catalytic activity of PQQ-ASD. Furthermore, the Km value for d-glucose of PaeASD s24-immobilized electrode decreased to approximately 1/3 that of the wild-type-immobilized electrode. These results indicate that the PaeASD s24 mutant is an excellent catalyst for potential bioelectrodevice applications.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"398 ","pages":"Pages 11-17"},"PeriodicalIF":4.1,"publicationDate":"2024-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142755169","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