Protein expression and purification最新文献

{"title":"Screening and functional characterization of nanobodies targeting the transferrin receptor","authors":"Yuting Ding ,&nbsp;Li Jia ,&nbsp;Qifubo Geng ,&nbsp;Yan Liu ,&nbsp;Shaojue Guo ,&nbsp;Shuaiying Zhao ,&nbsp;Yingying Kong ,&nbsp;Quanfang Jin ,&nbsp;Guangxu Xu ,&nbsp;Jianfeng Xu","doi":"10.1016/j.pep.2025.106702","DOIUrl":"10.1016/j.pep.2025.106702","url":null,"abstract":"<div><div>The transferrin receptor (TfR1) mediates the cellular uptake of iron and other molecules, playing a vital role in hematology and tumor growth. Nanobodies (NBs) targeting TfR1 offer promising therapeutic potential due to their small size, high specificity and stability. However, rapid identification of effective nanobodies remains challenging.In this study, the truncated extracellular fragment of human TfR1 was expressed in a prokaryotic system and purified. Immunized camelids provided a source for nanobody libraries, which were screened using phage display and high-throughput strategies to identify candidates with specific TfR1 binding.NB 2D7 with nanomolar-level dissociation constants (KD) were successfully identified.The analysis of Cell Counting Kit-8(CCK8) experiments indicates that the combined treatment of NB2D7 with FeCl₃ can reduce the survival rate of LoVo cells.This research establishes an efficient platform for anti-TfR1 nanobody screening and highlights the therapeutic potential of these nanobodies in cancer treatment and iron metabolism disorders.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"231 ","pages":"Article 106702"},"PeriodicalIF":1.4,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143634403","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}

{"title":"Expression of recombinant swine ferritin heavy chain with enhanced solubility in Escherichia coli and simplified purification of ferritin nanoparticles","authors":"M.S.B.W.T.M. Nipuna Sudaraka Tennakoon ,&nbsp;Kyoung-Ho Lee ,&nbsp;Hyun-Jin Shin","doi":"10.1016/j.pep.2025.106700","DOIUrl":"10.1016/j.pep.2025.106700","url":null,"abstract":"<div><div>Ferritin is a versatile biomolecule used in various medical applications such as drug delivery, vaccines, biological imaging, and diagnostics. The purity and concentration of the ferritin nanoparticles are crucial for achieving excellent outcomes. In this study, we expressed and purified the recombinant swine ferritin heavy chain (rsFTH) as a new candidate for recombinant ferritin nanoparticles. We generated two types of plasmids that can express rsFTH in mammalian and prokaryotic systems. The myc-tagged rsFTH expressed in the mammalian system was purified and ferritin nanoparticles were validated using dynamic light scattering (DLS) and transmission electron microscopy (TEM). A prokaryotic expression system was used to produce rsFTH on a large scale. Protein expression was optimized in Escherichia coli BL21 under varying temperatures and IPTG conditions, and solubility was enhanced by incubation at 25 °C for 18–22 h in auto-induction media, resulting in approximately &gt;50 % protein content in the soluble fraction compared with the pellet. Protein purification was achieved using His-tag affinity chromatography and dialysis with Tris-HCl buffer, yielding adequately pure rsFTH without any apparent protein aggregates. SDS-PAGE and Western blot analysis confirmed the expected molecular weight of rsFTH, and Native-PAGE demonstrated polymerization into higher molecular weight forms. Particle size analysis of purified rsFTH revealed a mean diameter of 15.5 nm, with transmission electron microscopy (TEM) imaging confirming spherical ferritin particles with an iron core. These results suggest that rsFTH can be efficiently expressed and purified in both mammalian and bacterial systems, and has potential applications in nanotechnology and biotechnology.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"231 ","pages":"Article 106700"},"PeriodicalIF":1.4,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143628203","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}

{"title":"Rational designation and characterization of a novel humanized collagen capable of self-assembling into triple helix and fibrils with D-period","authors":"Qiexin Chen ,&nbsp;Yao Zhang ,&nbsp;Yuxiang Zhang ,&nbsp;Xiao Han ,&nbsp;Luyao Zhang ,&nbsp;Huan Meng ,&nbsp;Jian Luo ,&nbsp;Rong Yu ,&nbsp;Chun Zhang ,&nbsp;Yongdong Liu","doi":"10.1016/j.pep.2025.106698","DOIUrl":"10.1016/j.pep.2025.106698","url":null,"abstract":"<div><div>The triple helix and D-period are distinctive features of native collagen, crucial for its physicochemical properties and bioactivities. However, developing recombinant humanized collagen with D-period features remains elusive. Here, we present a strategy for preparing a novel recombinant humanized collagen using a ‘charged-hydrophobic-charged amino acid’ sequence with the capacity of self-assembling. The hydrophobic amino acids in the middle region are believed to be crucial for the triple helix formation while the charged amino acids at the C- and N-terminal drive the triple-helix to self-assemble into higher-order structures like fibrils, with D-period formation during this process. To prove this concept, the particular fragment of Gly1059-Ala1103 of human type III collagen, featuring arginine (R), lysine (K), aspartic acid (D), and glutamic acid (E)-rich termini and a Glycine-Proline-Alanine (G-P-A) central motif, was selected and repeated to construct a recombinant humanized collagen, designated as rhCL04. This construct successfully formed hierarchical structures, including triple helices, rod-like fibrils, and hydrogels, exhibiting a distinct 10 nm D-period across a broad pH range from 4 to 10. Additionally, cell adhesion and biocompatibility were confirmed using L929 mouse fibroblast cells, demonstrating the ability to promote cell adhesion activity and no significant cytotoxicity. Our study provides valuable insights into the self-assembling mechanisms of native collagens. Moreover, these results highlight the efficacy of this strategy in producing recombinant humanized collagen with collagen-like characteristics. The simplicity and versatility of the approach, combined with the excellent self-assembling properties and biological activity of rhCL04, underscore its potential for biomaterial production.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"230 ","pages":"Article 106698"},"PeriodicalIF":1.4,"publicationDate":"2025-03-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143586757","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}

{"title":"High-purity preparation and biophysical characterization of OX40-TC in lipid nanodiscs","authors":"Hao Wang ,&nbsp;Xiaohui Zhang ,&nbsp;Cunlong Yin ,&nbsp;Lu Liu ,&nbsp;Liang Chen ,&nbsp;Hengfang Tang ,&nbsp;Bo Wu ,&nbsp;Junfeng Wang","doi":"10.1016/j.pep.2025.106697","DOIUrl":"10.1016/j.pep.2025.106697","url":null,"abstract":"<div><div>Tumor necrosis factor receptor superfamily member 4 (OX40) is essential for the activation and maintenance of T cell immune function because of its recognition and regulation in signal transduction, therefore understanding the structure and dynamics of the membrane protein OX40 in a near-native membrane environment is critical for elucidating its functions. However, efforts to prepare OX40 in a stable, functional form and reconstitute it into a membrane-mimicking lipid system face persistent challenges, limiting progress in its structural and functional characterization. Here, we developed an efficient method for the expression in <em>E. coli</em> and purification of the N-terminal truncated transmembrane and cytoplasmic domains of OX40 (OX40-TC) using a TrpLE fusion system. High-purity OX40-TC was achieved and successfully reconstituted into membrane scaffold protein (MSP)-nanodiscs for the first time. Characterization through size-exclusion chromatography, dynamic light scattering, and transmission electron microscopy confirmed their stability and homogeneity. This study provides a reliable platform for investigating the structure and function of OX40-TC, advancing our understanding of OX40-mediated signaling and its therapeutic potential.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"230 ","pages":"Article 106697"},"PeriodicalIF":1.4,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143551522","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}

{"title":"Mini review for niche downstream processes","authors":"Tsutomu Arakawa ,&nbsp;Daisuke Ejima ,&nbsp;Yui Tomioka ,&nbsp;Chiaki Sakuma ,&nbsp;Teruo Akuta","doi":"10.1016/j.pep.2025.106690","DOIUrl":"10.1016/j.pep.2025.106690","url":null,"abstract":"<div><div>We review here several niche downstream purification processes that are not covered by other articles in this special issue. The first is the use of activated carbon to capture contaminants and clarify culture medium for purification of proteins, including antibodies. Flow-through operation of the activated carbon filter showed over 80 % recovery of antibodies with 10--fold reduction of host cell proteins and effective lipopolysaccharide and virus removal. The second is salt or arginine-tolerant column chromatography, in which the respective resins, such as hydroxyapatite and mixed-mode resins, can bind proteins in the presence of salt or arginine at high concentrations. Effective use of arginine was also suggested in size exclusion and affinity chromatography. The last is the isolation of proteins using gel electrophoresis and simple extraction procedure. These technologies offer simple and cost-effective methods for purifying proteins and protein complexes in the native state.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"230 ","pages":"Article 106690"},"PeriodicalIF":1.4,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143465114","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}

{"title":"Expression and purification of recombinant glutaredoxin 1 and protection against oxidative stress injury during cerebral ischemia-reperfusion injury","authors":"Zi-teng Li,&nbsp;Tong Lin,&nbsp;Yu Sun,&nbsp;Xin-yi Wang,&nbsp;Yi-xuan Yang,&nbsp;Li Gan,&nbsp;Jia-ming Xu,&nbsp;Xu-ting Wei,&nbsp;Huang-qing Zhu,&nbsp;Wei-chun Zhao,&nbsp;Zhen-hong Zhu","doi":"10.1016/j.pep.2025.106689","DOIUrl":"10.1016/j.pep.2025.106689","url":null,"abstract":"<div><div>Glutaredoxin (Grx) is a small molecular protein widely found in both prokaryotes and eukaryotes, serving various biological functions, including participation in redox reactions and exerting anti-apoptotic effects[1]. To evaluate the protective effect of recombinant Grx1 against oxidative stress, we constructed the pET-30a (+)/Grx1 recombinant plasmid and performed soluble expression and purification of the recombinant Grx1. In vitro experiments, including ABTS and DPPH radical scavenging assays, showed that recombinant Grx1 has significant antioxidant activity. Reactive oxygen species detection revealed that the levels of reactive oxygen species in the Grx1 treatment group decreased by 33.01 % compared to the H₂O₂ group. Flow cytometry analyses indicated that the number of apoptotic cells in the Grx1 treatment group decreased by 23.51 % relative to the H₂O₂ group. Additionally, qRT-PCR analysis showed that Grx1 significantly reduced the expression levels of genes such as IL-1β, TNF-α, IL-6, and caspase-3 in PC12 cells. <em>In vivo</em>, recombinant Grx1 was utilized to treat cerebral ischemia-reperfusion injury (CIRI). Histological staining revealed that recombinant Grx1 significantly mitigated hippocampal tissue damage. Western blotting analysis demonstrated that Grx1 can reduce neuronal apoptosis following CIRI by decreasing Bax expression while increasing Bcl-2 expression. Furthermore, Grx1 was shown to modulate the HO-1/Nrf2 signaling pathway by elevating the expression of Nrf2 and HO-1. In summary, this study successfully overexpressed biologically active Grx1 in <em>E</em>. <em>coli,</em> and confirms that recombinant Grx1 exhibits remarkable antioxidant activity in both <em>in vitro</em> and <em>in vivo</em> experiments, effectively alleviating oxidative stress damage associated with ischemic stroke.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"230 ","pages":"Article 106689"},"PeriodicalIF":1.4,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143459212","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}

{"title":"StxB fusion strategy for immunogenic enhancement: Recombinant expression of AcAMP antimicrobial peptide","authors":"Ehsan Zamani ,&nbsp;Jamil Zargan ,&nbsp;Hossein Honari ,&nbsp;Hani Keshavarz Alikhani","doi":"10.1016/j.pep.2025.106688","DOIUrl":"10.1016/j.pep.2025.106688","url":null,"abstract":"<div><h3>Background and objectives</h3><div>Polyclonal antibodies are essential for identifying compounds like peptides. However, many peptides exhibit low immunogenicity, resulting in reduced antibody yields. To address this, fusing peptides with immunogenic fusion proteins has been proposed. This study aimed to enhance the immunogenicity of the AcAMP peptide by fusing it with StxB as a fusion protein, producing the recombinant construct in <em>Escherichia coli,</em> and comparing its antibody titration to recombinant AcAMP in mice.</div></div><div><h3>Methods</h3><div>The <em>acamp</em> gene, containing BamHI and SalI restriction sites, was amplified from the pUC57 plasmid via PCR and cloned into the pET28a (+)-StxB expression vector. The pET28a (+)-AcAMP construct was prepared as previously described. Both constructs were expressed in <em>E. coli</em> BL21 (DE3) cells, induced with IPTG, and purified using nickel affinity chromatography. Recombinant proteins were confirmed by SDS-PAGE and Western blotting. Mice were immunized with purified proteins, and serum IgG titration was assessed using indirect ELISA.</div></div><div><h3>Results</h3><div>PCR amplification and enzymatic digestion verified the successful construction of the pET28a (+)-<em>StxB-acamp</em> vector. SDS-PAGE and Western blotting identified recombinant AcAMP and AcAMP-StxB proteins at 9 and 17 kDa, respectively. ELISA revealed significantly higher antibody titers for the recombinant AcAMP-StxB protein compared to recombinant AcAMP.</div></div><div><h3>Conclusion</h3><div>Fusing the AcAMP peptide with the StxB protein enhanced immunogenicity and increased antibody production. This approach may improve expression conditions and immunogenicity for peptides of this family, advancing their use in identification studies.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"229 ","pages":"Article 106688"},"PeriodicalIF":1.4,"publicationDate":"2025-02-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143429167","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}

{"title":"Production of recombinant coiled coil silk proteins for materials synthesis","authors":"Patrick J. Shilling ,&nbsp;Luisa Pontes-Braz ,&nbsp;Lachlan Mitchell ,&nbsp;Linda Howell ,&nbsp;Prem Veneer ,&nbsp;Srinivasan Jayashree ,&nbsp;Laura A. Castelli ,&nbsp;Tam Pham ,&nbsp;Louis Lu ,&nbsp;Bei Wang ,&nbsp;K.Y Benjamin Yeo ,&nbsp;Surekha Nimma ,&nbsp;Lyndall Briggs ,&nbsp;Caitlin Johnston ,&nbsp;Michelle Michie ,&nbsp;Tara D. Sutherland","doi":"10.1016/j.pep.2025.106683","DOIUrl":"10.1016/j.pep.2025.106683","url":null,"abstract":"<div><div>Rational design of fundamentally new advanced materials would be facilitated by availability of polymers with controlled monomer sequence. Recombinant proteins offer polymers with controlled monomer sequence but are underrepresented in material science, in part because suitable proteins cannot be produced at commercial levels in recombinant systems. The silk proteins of honeybees fulfil the requirements for rational materials design and can be produced at commercially viable levels. In this study we compare recombinant expression of these silks in bacteria, yeast and insect cells to identify the most suitable method of silk protein production. Yeast and insect cell lines are unlikely to be suitable expression platforms for these silks as the recombinant proteins were degraded, expression levels were low or absent, and host cell protein levels were high. We confirm that expression into <em>E. coli</em> inclusion bodies using defined media offers high level expression and to date is the best expression system for these proteins.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"229 ","pages":"Article 106683"},"PeriodicalIF":1.4,"publicationDate":"2025-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143374397","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"High-affinity nanobodies targeting IL-12B for the detection of fluorescence resonance energy transfer","authors":"Jing Hu ,&nbsp;Wenxuan Feng ,&nbsp;Jianchuan Wen ,&nbsp;Siyu Zhou ,&nbsp;Zengchao Sun ,&nbsp;Shuaiying Zhao ,&nbsp;Shaojue Guo ,&nbsp;Hui Wang ,&nbsp;Yong Geng","doi":"10.1016/j.pep.2025.106681","DOIUrl":"10.1016/j.pep.2025.106681","url":null,"abstract":"<div><h3>Aims</h3><div>IL-12B, a subunit of the IL-23 family of cytokines, plays a crucial role in various diseases such as viral infections, autoimmune disorders, and tumors. This study aimed to identify high-affinity nanobodies that bind to distinct epitopes of IL-12B to and assess their potential for therapeutic and diagnostic applications, particularly through fluorescence resonance energy transfer(FRET) to evaluate their ability to target IL-12B.</div></div><div><h3>Methods</h3><div>IL-12B protein was expressed in eukaryotic cells and used to immunize camels to induce an immune response. Camel-derived anti-IL-12B nanobodies were isolated and screened via phage display to identify those with high specificity and affinity for IL-12B. Binding affinity and epitope interactions were further analyzed using high-performance liquid chromatography (HPLC) and ForteBio Octet assays. A FRET-based assay was developed to evaluate protein interactions for precise therapeutic targeting.</div></div><div><h3>Results</h3><div>Several high-affinity nanobodies targeting IL-12B were successfully generated. These nanobodies exhibited strong binding to various epitopes of IL-12B. Screening by HPLC and ForteBio Octet confirmed their high specificity and affinity, while fluorescence analysis demonstrated efficient energy transfer between thenanobodies, indicating successful interactions.</div></div><div><h3>Conclusions</h3><div>This study identified high-affinity nanobodies against IL-12B and used FRET to characterize their interactions. These nanobodies show promise for therapeutic potential targeting IL-12B-related diseases, including viral infections, autoimmune disorders, and cancer. However, further clinical studies are needed to fully explore their potential for diagnostic and therapeutic applications.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"229 ","pages":"Article 106681"},"PeriodicalIF":1.4,"publicationDate":"2025-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143374386","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}

{"title":"Expression and biochemical characterization of a novel NAD+-dependent xylitol dehydrogenase from the plant endophytic fungus Trichoderma gamsii","authors":"Shuping Fei ,&nbsp;Wenxiu Hu ,&nbsp;Jingwen Shu ,&nbsp;Ruirui Zhao ,&nbsp;Jiatong Zhao ,&nbsp;Mengwei Jiang ,&nbsp;Wenwen Wu ,&nbsp;Chaoqun Lian ,&nbsp;Wanggang Tang","doi":"10.1016/j.pep.2025.106687","DOIUrl":"10.1016/j.pep.2025.106687","url":null,"abstract":"<div><div>Xylitol dehydrogenase (XDH; EC 1.1.1.9), encoded by the <em>XYL2</em> gene, is a key enzyme in the fungal xylose metabolic pathway. In this work, a putative XDH from the plant endophytic fungus <em>Trichoderma gamsii</em> (TgXDH) was hetero-expressed in <em>Escherichia coli</em> BL21(DE3), purified to the homogeneity, and biochemically characterized. Sequence analysis revealed that TgXDH is 363 amino acids long and belongs to the zinc-containing medium-chain alcohol dehydrogenase superfamily. The size-exclusion chromatography analysis and SDS-PAGE showed that the purified recombinant TgXDH had a native molecular mass of ∼155 kDa and was composed of four identical subunits of molecular mass of ∼39 kDa. The optimum temperature and pH of this enzyme were 25 °C and pH 9.5, respectively. Kinetic analysis showed that it is an NAD⁺-dependent enzyme that has a polyol substrate preference (based on <em>k</em>_cat/<em>K</em>_m) in the order xylitol &gt; ribitol ≈ <span>d</span>-sorbitol. The <em>K</em>_m values for NAD⁺ with these three polyols ranged from 0.23 to 0.70 mM. Moreover, TgXDH showed high substrate affinities as compared to most of its homologs. The <em>K</em>_m values for xylitol, ribitol, and <span>d</span>-sorbitol were 5.23 ± 0.68 mM, 8.01 ± 1.22 mM, and 12.34 ± 1.37 mM, respectively. Collectively, the results will contribute to understanding the biochemical properties of a novel XDH from the filamentous fungi and provide a promising XDH for industrial production of ethanol.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"229 ","pages":"Article 106687"},"PeriodicalIF":1.4,"publicationDate":"2025-02-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143365722","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}