Journal of bioscience and bioengineering最新文献_第2页

Amphiphilic phospholipid polymers as a cryoprotectant for vitrification and nanowarming of rat livers. 两亲磷脂聚合物作为玻璃化和纳米加热大鼠肝脏的低温保护剂

IF 2.3 4区生物学

Journal of bioscience and bioengineering Pub Date : 2024-10-24 DOI: 10.1016/j.jbiosc.2024.10.003

Masahiro Kaneko, Natsumi Takizawa, Taisei Wakabayashi, Hidenori Kaneoka, Akira Ito

{"title":"Amphiphilic phospholipid polymers as a cryoprotectant for vitrification and nanowarming of rat livers.","authors":"Masahiro Kaneko, Natsumi Takizawa, Taisei Wakabayashi, Hidenori Kaneoka, Akira Ito","doi":"10.1016/j.jbiosc.2024.10.003","DOIUrl":"https://doi.org/10.1016/j.jbiosc.2024.10.003","url":null,"abstract":"<p><p>Liver biobanking is a promising approach that saves the lives of patients with end-stage liver disease. Cryopreservation based on vitrification enables semi-permanent organ preservation, contributing to overcome the shortage of donors for liver transplants. A technical challenge in cryopreservation of transplantable organs lies in thawing methodology, and conventional convective warming cannot maintain the glassy state during thawing because of the large temperature gradient between the inner and outer parts of the organs, leading to ice formation and damage of cells in the organ. Nanowarming, in which magnetic nanoparticles are dispersed in a vitrification solution and heated by exposure of alternating magnetic field, can achieve uniform and rapid heating of organs. Herein, we report that amphiphilic phospholipid polymers composed of 2-methacryloyloxyethyl phosphorylcholine and n-butyl methacrylate can function as a cryoprotectant for nanowarming. The amphiphilic phospholipid polymers enhanced the viability of primary rat hepatocytes after vitrification. Moreover, the polymers enhanced the dispersion stability of magnetic nanoparticles in vitrification solution, and the perfusion of the vitrification solution with magnetic nanoparticles into rat livers through portal vein provided uniform distribution of the nanoparticles in the liver. After perfusion, the vitrified liver was successfully thawed rapidly and uniformly by nanowarming, which maintained tissue integrity and cell viability.</p>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":" ","pages":""},"PeriodicalIF":2.3,"publicationDate":"2024-10-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142501013","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Application of a low acetate-producing strain of Tetragenococcus halophilus to soy sauce fermentation. 在酱油发酵中应用嗜卤四源球菌低醋酸盐菌株。

IF 2.3 4区生物学

Journal of bioscience and bioengineering Pub Date : 2024-10-18 DOI: 10.1016/j.jbiosc.2024.09.007

Keita Higuchi, Yuya Nukagawa, Takura Wakinaka, Jun Watanabe, Yoshinobu Mogi

{"title":"Application of a low acetate-producing strain of Tetragenococcus halophilus to soy sauce fermentation.","authors":"Keita Higuchi, Yuya Nukagawa, Takura Wakinaka, Jun Watanabe, Yoshinobu Mogi","doi":"10.1016/j.jbiosc.2024.09.007","DOIUrl":"https://doi.org/10.1016/j.jbiosc.2024.09.007","url":null,"abstract":"<p><p>In soy sauce brewing, the halophilic lactic acid bacterium, Tetragenococcus halophilus is used as a fermentation starter and contributes to the taste and aroma of soy sauce, mainly by producing lactate. By lowering the pH of the soy sauce mash, lactate serves as a suitable growth environment for the halotolerant yeast Zygosaccharomyces rouxii. Acetate, which is produced by T. halophilus via the citrate metabolic pathway, is a critical growth inhibitory factor for Z. rouxii. Therefore, a T. halophilus strain that lacks acetate production could be an ideal fermentation starter to enhance ethanol production. In this study, we obtained a derivative of T. halophilus containing an insertion sequence in citC, which is an essential gene for citrate metabolism, and validated its performance as a soy sauce fermentation starter. The derivative neither metabolized citrate nor produced excessive acetate in soy sauce mash, resulting in vigorous alcohol fermentation by Z. rouxii. This study provides insights into the application of a low acetate-producing strain of T. halophilus as a starter to produce soy sauce with high alcohol content and low sour aroma.</p>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":" ","pages":""},"PeriodicalIF":2.3,"publicationDate":"2024-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142466295","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Evaluation of induced pluripotent stem cell differentiation into neural progenitor cell using Raman spectra derived from extracellular vesicles in culture supernatants. 利用培养上清液中细胞外囊泡的拉曼光谱评估诱导多能干细胞向神经祖细胞分化的情况。

IF 2.3 4区生物学

Journal of bioscience and bioengineering Pub Date : 2024-10-16 DOI: 10.1016/j.jbiosc.2024.09.004

Kakuro Hirai, Hikaru Saito, Midori Kato, Masaharu Kiyama, Hiroko Hanzawa, Atsushi Nakane, Sayaka Sekiya, Kenji Yoshida, Akiyoshi Kishino, Atsushi Ikeda, Toru Kimura, Jun Takahashi, Shizu Takeda

{"title":"Evaluation of induced pluripotent stem cell differentiation into neural progenitor cell using Raman spectra derived from extracellular vesicles in culture supernatants.","authors":"Kakuro Hirai, Hikaru Saito, Midori Kato, Masaharu Kiyama, Hiroko Hanzawa, Atsushi Nakane, Sayaka Sekiya, Kenji Yoshida, Akiyoshi Kishino, Atsushi Ikeda, Toru Kimura, Jun Takahashi, Shizu Takeda","doi":"10.1016/j.jbiosc.2024.09.004","DOIUrl":"https://doi.org/10.1016/j.jbiosc.2024.09.004","url":null,"abstract":"<p><p>Non-invasive cell culture monitoring technology is crucial to improve the manufacturing efficiency of cell products. We have found that extracellular vesicles (EVs) are secreted into the culture supernatants in the differentiation process from human induced pluripotent stem cells (iPSCs) to dopaminergic progenitor cells, and that the composition of EVs changes in accordance with the differentiation processes. In this study, we hypothesized that it is possible to evaluate the cultured cellular states by detecting compositional changes of EVs secreted from cultured cells with label-free Raman spectroscopy in a non-invasive manner. Therefore, Raman signal analysis derived from EV fractions isolated from culture supernatants throughout the differentiation process was conducted. iPSCs cultures were simultaneously implemented under a standard condition (control) and an artificial deviation condition inducing reductions in pluripotency by depleting FGF2 in culture medium (-FGF2), which is indispensable for maintaining the pluripotency. Subsequently, the differentiation step was conducted for each iPSCs culture under the same condition. As a result, it was found that under -FGF2, the expression level of the pluripotency marker NANOG decreased compared to that of the control and correlated with the identification results based on Raman signals with a correlation coefficient of 0.77. Lipid-derived Raman signals were extracted as identification factors, suggesting that changes in the lipid component of EV occur depending on the cellular states. From the above, we have found that the change in composition of EVs in the culture supernatant by detecting Raman signals would be a monitoring index of the cellular state of differentiation and pluripotency.</p>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":" ","pages":""},"PeriodicalIF":2.3,"publicationDate":"2024-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142466297","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Bioconversion of eicosapentaenoic acid into 5S,15S- and 5R,15R-dihydroxyeicosapentaenoic acids by double-dioxygenating 15S- and 15R-lipoxygenases. 通过 15S- 和 15R- 脂氧合酶将二十碳五烯酸生物转化为 5S、15S- 和 5R、15R- 二羟基二十碳五烯酸。

IF 2.3 4区生物学

Journal of bioscience and bioengineering Pub Date : 2024-10-14 DOI: 10.1016/j.jbiosc.2024.09.002

Jin Lee, Hyun-Ah Park, Kyung-Chul Shin, Deok-Kun Oh

{"title":"Bioconversion of eicosapentaenoic acid into 5S,15S- and 5R,15R-dihydroxyeicosapentaenoic acids by double-dioxygenating 15S- and 15R-lipoxygenases.","authors":"Jin Lee, Hyun-Ah Park, Kyung-Chul Shin, Deok-Kun Oh","doi":"10.1016/j.jbiosc.2024.09.002","DOIUrl":"https://doi.org/10.1016/j.jbiosc.2024.09.002","url":null,"abstract":"<p><p>Resolvin E series (Rvs), such as RvE4 (5S,15S-dihydroxyeicosapentaenoic acid) and its stereoselective enantiomer (5R,15R-dihydroxyeicosapentaenoic acid), play an important role in promoting the resolution of inflammation and are derived from eicosapentaenoic acid (EPA) by M2 macrophage in human. However, they have been synthesized using expensive and inefficient chemical methods. Here, we performed efficient quantitative production of RvE4 and its enantiomer from EPA using Escherichia coli expressing double-dioxygenating 15S-lipoxygenase (15S-LOX) from Archangium violaceum and double-dioxygenating 15R-LOX from Sorangium cellulosum, respectively, with solvent, polymer, and adsorbent resin. The cell density, substrate concentration, solvent types and concentrations, polymer types and concentrations, and resin concentration were optimized for the enhanced bioconversion of EPA into RvE4 and its enantiomer. Under the optimized conditions, A. violaceum 15S-LOX and S. cellulosum 15R-LOX expressed in E. coli converted 6.0 mM EPA into 4.3 mM (1.44 g/L) RvE4 and 5.8 mM (1.94 g/L) RvE4 enantiomer in 60 min, with productivities of 4.3 and 5.8 mM/h and molar conversions of 72% and 97%, respectively. To date, these are the highest concentrations, productivities, and conversions of RvE4 and its enantiomer. The concentrations of RvE4 and its enantiomer obtained from the conversion of EPA with solvent, polymer, and resin were 2.5- and 3.2-fold higher than those without the additives, respectively.</p>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":" ","pages":""},"PeriodicalIF":2.3,"publicationDate":"2024-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142466296","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Positive impact of pyrocarbon and mechanical loading on cartilage-like tissue synthesis in a scaffold-free process. 在无支架工艺中，热碳和机械负载对软骨样组织合成的积极影响。

IF 2.3 4区生物学

Journal of bioscience and bioengineering Pub Date : 2024-10-11 DOI: 10.1016/j.jbiosc.2024.09.005

Imbert De Gaudemaris, Amira Hannoun, Rémy Gauthier, Nina Attik, Leyre Brizuela, Saida Mebarek, Michel Hassler, Carole Bougault, Ana-Maria Trunfio-Sfarghiu

{"title":"Positive impact of pyrocarbon and mechanical loading on cartilage-like tissue synthesis in a scaffold-free process.","authors":"Imbert De Gaudemaris, Amira Hannoun, Rémy Gauthier, Nina Attik, Leyre Brizuela, Saida Mebarek, Michel Hassler, Carole Bougault, Ana-Maria Trunfio-Sfarghiu","doi":"10.1016/j.jbiosc.2024.09.005","DOIUrl":"https://doi.org/10.1016/j.jbiosc.2024.09.005","url":null,"abstract":"<p><p>Aiming to build a tissue analogue engineered cartilage from differentiated chondrocytes, we investigated the potential of a pyrocarbon (PyC)-based and scaffold-free process, under mechanical stimulation. PyC biomaterial has shown promise in arthroplasty and implant strategies, and mechanical stimulation is recognized as an improvement in regeneration strategies. The objective was to maintain the cell phenotype to produce constructs with cartilage-like matrix composition and mechanical properties. Primary murine chondrocytes were deposited in drop form between two biomaterial surfaces expanded to 500 μm and a uniaxial cyclic compression was applied thanks to a handmade tribo-bioreactor (0.5 Hz, 100 μm of amplitude, 17 days). Histology and immunohistochemistry analysis showed that PyC biomaterial promoted expression of cartilage-like matrix components (glycosaminoglycans, type II collagen, aggrecan). Importantly, constructs obtained in dynamic conditions were denser and showed a cohesive and compact shape. The most promising condition was the combined use of PyC and dynamic stimulation, resulting in constructs of low elasticity and high viscosity, thus with an increased damping factor. We verified that no calcium deposits were detectable and that type X collagen was not expressed, suggesting that the cells had not undergone hypertrophic maturation. While most studies focus on the comparison of different biomaterials or on the effect of different mechanical stimuli separately, we demonstrated the value of combining the two approaches to get as close as possible to the biological and mechanical qualities of natural hyaline articular cartilage.</p>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":" ","pages":""},"PeriodicalIF":2.3,"publicationDate":"2024-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142466308","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Optimization of bacteriophage propagation in high-yield continuous culture (cellstat) meeting the constraints of industrial manufacturing processes 优化噬菌体在高产连续培养（cellstat）中的繁殖，以满足工业生产过程的限制。

IF 2.3 4区生物学

Journal of bioscience and bioengineering Pub Date : 2024-10-05 DOI: 10.1016/j.jbiosc.2024.09.001

Céleste Caffin , Lhéa Milhamont , Eva Duriez , Agathe Hembert , Pauline Huzet , Camille Lerouge , Marie Deblieck , Denis Watier

{"title":"Optimization of bacteriophage propagation in high-yield continuous culture (cellstat) meeting the constraints of industrial manufacturing processes","authors":"Céleste Caffin , Lhéa Milhamont , Eva Duriez , Agathe Hembert , Pauline Huzet , Camille Lerouge , Marie Deblieck , Denis Watier","doi":"10.1016/j.jbiosc.2024.09.001","DOIUrl":"10.1016/j.jbiosc.2024.09.001","url":null,"abstract":"<div><div>The growing use of bacteriophages in the fields of agriculture, agri-food, veterinary treatments, and medicine involves the quantitative production of these bacteriophages. In this study, we propose a bacteriophage production protocol that can easily be transposed to industry. We used a cellstat production system because the latest studies have shown that it is the most suitable process for the production of phages due to volumetric productivity, safety (limitation of co-evolution), and flexibility (choice of growth rate criteria). Sizing of the assembly used makes it possible to extrapolate the results to industrial production. The production conditions are indicated precisely, which would allow manufacturers to adapt the protocol to their own equipment. We propose experimental conditions in order to obtain a stable <em>Escherichia coli</em> population, qualitatively and over time, and production of high-titer T7 bacteriophages. The optimized production conditions (yield, cost and simplicity of the process) are: a buffered peptone water medium concentration of 11 g L<sup>−1</sup> and a dilution rate of 1.6 h<sup>−1</sup>. Under these conditions, we obtained a production of 7.35×10<sup>16</sup> plaque-forming units (PFU) L<sup>−1</sup> day<sup>−1</sup> with a concentration of 9.8×10<sup>12</sup> PFU mL<sup>−1</sup>. The strength of this work lies in its focus on industrial applicability.</div></div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"138 6","pages":"Pages 507-514"},"PeriodicalIF":2.3,"publicationDate":"2024-10-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142377847","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Effect of sodium metabisulfite treatment and storage condition on metabolic profile of young coconut (Cocos nucifera L.) 焦亚硫酸钠处理和储存条件对椰子（Cocos nucifera L.）新陈代谢的影响

IF 2.3 4区生物学

Journal of bioscience and bioengineering Pub Date : 2024-09-28 DOI: 10.1016/j.jbiosc.2024.08.002

Della Rahmawati , Mary Faith Yamballa Adan , Muhammad Maulana Malikul Ikram , Marvin Nathanael Iman , Eiichiro Fukusaki , Sastia Prama Putri

{"title":"Effect of sodium metabisulfite treatment and storage condition on metabolic profile of young coconut (Cocos nucifera L.)","authors":"Della Rahmawati , Mary Faith Yamballa Adan , Muhammad Maulana Malikul Ikram , Marvin Nathanael Iman , Eiichiro Fukusaki , Sastia Prama Putri","doi":"10.1016/j.jbiosc.2024.08.002","DOIUrl":"10.1016/j.jbiosc.2024.08.002","url":null,"abstract":"<div><div>Young coconuts (<em>Cocos nucifera</em> L.) used for export are trimmed to reduce their size and weight to lower transport costs. However, trimmed coconuts have a shorter shelf life due to microbial spoilage and surface discoloration caused by enzymatic browning. To minimize these effects, trimmed coconuts were dipped in an anti-browning agent, sodium metabisulfite (SMB), and stored under ambient conditions. However, there have been no reports on the effects of SMB treatment on metabolome changes in the flesh and water of young coconuts. Hence, this study investigated the metabolite changes in trimmed young coconuts after SMB treatment under different storage conditions using a gas chromatography (GC)/mass spectrometry (MS) metabolomic profiling approach. Tall young coconut samples were trimmed and treated with a 2% SMB solution for 5 min before storage at 25 °C or 4 °C for 2–4 weeks. Coconut flesh and water samples were collected after storage for 0, 2, and 4 weeks, and were subjected to GC–MS analysis. The results showed that the major metabolites affected by coconut deterioration were amino acids, sugars, and sugar alcohols. SMB treatment and/or refrigeration can help prevent metabolite changes in the flesh and water of young coconuts. In the future, improvements in storage conditions based on metabolite profiles should be explored.</div></div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"138 6","pages":"Pages 515-521"},"PeriodicalIF":2.3,"publicationDate":"2024-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142347274","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Enzymatically cross-linkable sulfated bacterial polyglucuronic acid as an affinity-based carrier of FGF-2 for therapeutic angiogenesis 可酶法交联硫酸化细菌聚葡萄糖醛酸是一种基于亲和力的 FGF-2 载体，可用于治疗性血管生成。

IF 2.3 4区生物学

Journal of bioscience and bioengineering Pub Date : 2024-09-28 DOI: 10.1016/j.jbiosc.2024.08.011

Ryota Goto , Shinji Sakai , Cédric Delattre , Emmanuel Petit , Redouan El Boutachfaiti , Masaki Nakahata

{"title":"Enzymatically cross-linkable sulfated bacterial polyglucuronic acid as an affinity-based carrier of FGF-2 for therapeutic angiogenesis","authors":"Ryota Goto , Shinji Sakai , Cédric Delattre , Emmanuel Petit , Redouan El Boutachfaiti , Masaki Nakahata","doi":"10.1016/j.jbiosc.2024.08.011","DOIUrl":"10.1016/j.jbiosc.2024.08.011","url":null,"abstract":"<div><div>The fibroblast growth factor-2 (FGF-2) is a critical protein for biological processes such as angiogenesis and tissue regeneration. Recently, hydrogels based on semi-synthetic sulfated polysaccharides have been developed for the controlled delivery of FGF-2. These affinity-based FGF-2 carriers utilizing hydrogels based on sulfated polysaccharides enable sustained delivery of FGF-2, yet choice of materials is limited. Here, we demonstrate a novel synthetic sulfated polysaccharide-based hydrogel based on bacterial polyglucuronic acid (PGU). We synthesized phenol-grafted sulfated PGU (PGUS-Ph), an enzymatically cross-linkable PGU derivative that exhibited an enhanced affinity for FGF-2. The aqueous solution of PGUS-Ph, when combined with FGF-2, could be injected into affected sites and form a hydrogel in a minimally invasive manner. The FGF-2 released from the PGUS-Ph hydrogel induced blood vessel formation, as proven by a chick embryo-based angiogenesis assay. Our results indicate that the PGUS-Ph has the potential as an enzymatically cross-linkable and minimally invasively injectable affinity-based FGF-2 delivery system.</div></div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"138 6","pages":"Pages 541-547"},"PeriodicalIF":2.3,"publicationDate":"2024-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142347276","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Elucidation of d-allulose recognition mechanism in ketose 3-epimerase 阐明酮糖 3-酰亚胺酶中 d-阿洛糖的识别机制。

IF 2.3 4区生物学

Journal of bioscience and bioengineering Pub Date : 2024-09-23 DOI: 10.1016/j.jbiosc.2024.08.010

Masahiro Watanabe , Yusuke Nakamichi , Shohei Mine

{"title":"Elucidation of d-allulose recognition mechanism in ketose 3-epimerase","authors":"Masahiro Watanabe , Yusuke Nakamichi , Shohei Mine","doi":"10.1016/j.jbiosc.2024.08.010","DOIUrl":"10.1016/j.jbiosc.2024.08.010","url":null,"abstract":"<div><div><span>d</span>-Allulose is a low-calorie sweetener with multiple nutritional functions that can be produced through <span>d</span>-fructose isomerization by ketose 3-epimerase (KEase). <span>l</span>-Ribulose 3-epimerase from <em>Arthrobacter</em> <em>globiformis</em> (AgLRE) is one of the most important enzymes that produce <span>d</span>-allulose; however, its substrate recognition mechanism is unknown. In this study, the crystal structures of AgLRE and its complex with <span>d</span>-allulose and <span>d</span>-fructose were determined. Upon substrate binding, the hydrophobic residues around the active-site entrance move toward the bound substrate. A comparison of AgLRE and other KEase structures revealed that the substrate-binding residues are not the main factors responsible for its marked specificity for <span>d</span>-allulose and <span>d</span>-fructose, but the hydrophobicity of the active site pocket influences substrate recognition. Particularly, the two hydrophobic regions at the active site entrance are the regulatory elements that modulate substrate recognition by AgLRE. This study provides useful information for designing AgLRE to increase its affinity for <span>d</span>-allulose and <span>d</span>-fructose.</div></div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"138 6","pages":"Pages 488-494"},"PeriodicalIF":2.3,"publicationDate":"2024-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142347275","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Rapid one-pot human single nucleotide polymorphism genotyping platform with Cas13a nuclease 使用 Cas13a 核酸酶的单锅快速人类单核苷酸多态性基因分型平台。

IF 2.3 4区生物学

Journal of bioscience and bioengineering Pub Date : 2024-09-19 DOI: 10.1016/j.jbiosc.2024.08.003

Rui Lei , Xi-Peng Liu

{"title":"Rapid one-pot human single nucleotide polymorphism genotyping platform with Cas13a nuclease","authors":"Rui Lei , Xi-Peng Liu","doi":"10.1016/j.jbiosc.2024.08.003","DOIUrl":"10.1016/j.jbiosc.2024.08.003","url":null,"abstract":"<div><div>Single nucleotide polymorphism (SNP), as one of the key components of the genetic factors, is important for disease detection and early screening of hereditary diseases. Current SNP genotyping methods require laboratory instruments or long operating times. To facilitate the diagnosis of hereditary diseases, we developed a new method referred to as the LwaCas13a-based SNP genotyping platform (Cas13a platform), which is useful for detecting disease-related SNPs. We report a CRISPR/Cas13a-based SNP genotyping platform that couples recombinase-aided amplification (RAA), T7 transcription, and <em>Leptotrichia wadei</em> Cas13a (LwaCas13a) detection for simple and fast genotyping of human disease-related SNPs. We used this Cas13a platform to identify 17 disease-related SNPs, demonstrating that position 2 in gRNA is suitable for the introduction of additional mismatches to achieve high discrimination in genotyping across a wide range of SNP targets. The discrimination specificity of 17 SNPs was improved 3.0–35.1-fold after introducing additional mismatches at position 2 from the 5′-end. We developed a method, which has a lower risk of cross-contamination and operational complexity, for genotyping SNPs using human saliva samples in an one-pot testing that delivers results within 60 min. Compared to TaqMan probe qPCR, RFLP, AS-PCR and other SNP genotyping methods, the Cas13a platform is simple, rapid and reliable, expanding the applications of the CRISPR/Cas system in nucleic acid detection and SNP genotyping.</div></div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"138 6","pages":"Pages 469-477"},"PeriodicalIF":2.3,"publicationDate":"2024-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142288070","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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