Immobilized macromolecular enzymes effectively address critical challenges including environmental sensitivity, thermal instability, and non-reusability, thereby establishing the fundamental and practical significance of enzyme immobilization research. This study employed sodium alginate (SA), ZIF-8, and Bi-EA materials to immobilize Aspergillus carlsbadensis-derived epoxide hydrolase (AcEH). Comparative analysis revealed SA's superior immobilization efficiency. Material characterization via SEM, FTIR, and XRD was performed, followed by optimization of SA immobilization parameters. The immobilized enzyme maintained activity over 6 reuse cycles. Incorporating 0.5 wt% carboxymethyl cellulose (CMC) extended AcEH@SA-CMC reusability to 8 cycles. Magnetic modification through Fe3O4 powder incorporation yielded AcEH@SA-CMC- Fe3O4 hydrogel with enhanced recoverability. Results demonstrate SA's exceptional biocompatibility underpins its immobilization efficacy, while CMC co-entanglement creates a porous network facilitating mass transfer and mechanical reinforcement. Fe3O4 powder plays a promoting role in improving enzyme activity. This methodology not only evaluates immobilization matrices but establishes an optimized protocol for macromolecular AcEH immobilization.
{"title":"Sodium alginate-immobilized epoxide hydrolase: A multifaceted strategy for enhanced stability, reusability, and catalytic performance","authors":"Zongzhong Yu , Meinan Zhu , Xiao Gu , Zhongkun Wu , Peiqin Chen , Chunying Jin , Junning Zhao , Guangya Zhang , Wei Jiang","doi":"10.1016/j.jbiotec.2025.11.017","DOIUrl":"10.1016/j.jbiotec.2025.11.017","url":null,"abstract":"<div><div>Immobilized macromolecular enzymes effectively address critical challenges including environmental sensitivity, thermal instability, and non-reusability, thereby establishing the fundamental and practical significance of enzyme immobilization research. This study employed sodium alginate (SA), ZIF-8, and Bi-EA materials to immobilize <em>Aspergillus carlsbadensis</em>-derived epoxide hydrolase (AcEH). Comparative analysis revealed SA's superior immobilization efficiency. Material characterization via SEM, FTIR, and XRD was performed, followed by optimization of SA immobilization parameters. The immobilized enzyme maintained activity over 6 reuse cycles. Incorporating 0.5 wt% carboxymethyl cellulose (CMC) extended AcEH@SA-CMC reusability to 8 cycles. Magnetic modification through Fe<sub>3</sub>O<sub>4</sub> powder incorporation yielded AcEH@SA-CMC- Fe<sub>3</sub>O<sub>4</sub> hydrogel with enhanced recoverability. Results demonstrate SA's exceptional biocompatibility underpins its immobilization efficacy, while CMC co-entanglement creates a porous network facilitating mass transfer and mechanical reinforcement. Fe<sub>3</sub>O<sub>4</sub> powder plays a promoting role in improving enzyme activity. This methodology not only evaluates immobilization matrices but establishes an optimized protocol for macromolecular AcEH immobilization.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 84-95"},"PeriodicalIF":3.9,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145622129","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}
Pub Date : 2026-02-01Epub Date: 2025-12-06DOI: 10.1016/j.jbiotec.2025.12.003
Tao Liu , Yuzhang Zhu , Jiahui Wang , Kun Lin , Xuan Zhang , Huawei Ma , Chaonan Kong , Wei Cai , Lifang Yang , Mingguo Jiang
The development of natural product photosensitizers was a critical cornerstone for applying photodynamic antimicrobial technology in the food industry. The objective of this study was to fabricate complex of fisetin-β-cyclodextrin (FIT-β-CD), explore their photodynamic antimicrobial efficacy and mechanisms, and characterize it through infrared spectroscopy and X-ray diffraction. The results indicatde that this method can produce good dispersibility and water solubility of FIT-β-CD complex.The photodynamic impact of FIT-β-CD was assessed by examining alterations in reactive oxygen species (ROS) production.The results demonstrated that FIT-β-CD can rapidly generate ROS under UVA, with hydrogen peroxide production being 3.73 times higher than in dark conditions. The antimicrobial efficacy of FIT-β-CD against Escherichia coli O157:H7 (E.coli O157:H7) was evaluated based on bactericidal efficacy, cell membrane damage, DNA and protein damage, motility changes, biofilm formation, and extracellular polysaccharide production. In addition, gene expression analysis by RT-qPCR showed its significant inhibition on the two-component system (TCS) and quorum sensing system (QS). The results indicated that FIT-β-CD exhibited efficient, sensitive photodynamic, and antibacterial properties against E.coli O157:H7, making it a novel photosensitizer. These findings imply potential new applications of fisetin in the field of food antimicrobial agents.
{"title":"Antibacterial effects and mechanisms of fisetin-β-cyclodextrin complex under UVA light against Escherichia coli O157:H7","authors":"Tao Liu , Yuzhang Zhu , Jiahui Wang , Kun Lin , Xuan Zhang , Huawei Ma , Chaonan Kong , Wei Cai , Lifang Yang , Mingguo Jiang","doi":"10.1016/j.jbiotec.2025.12.003","DOIUrl":"10.1016/j.jbiotec.2025.12.003","url":null,"abstract":"<div><div>The development of natural product photosensitizers was a critical cornerstone for applying photodynamic antimicrobial technology in the food industry. The objective of this study was to fabricate complex of fisetin-β-cyclodextrin (FIT-β-CD), explore their photodynamic antimicrobial efficacy and mechanisms, and characterize it through infrared spectroscopy and X-ray diffraction. The results indicatde that this method can produce good dispersibility and water solubility of FIT-β-CD complex.The photodynamic impact of FIT-β-CD was assessed by examining alterations in reactive oxygen species (ROS) production.The results demonstrated that FIT-β-CD can rapidly generate ROS under UVA, with hydrogen peroxide production being 3.73 times higher than in dark conditions. The antimicrobial efficacy of FIT-β-CD against <em>Escherichia coli</em> O157:H7 (<em>E.coli</em> O157:H7) was evaluated based on bactericidal efficacy, cell membrane damage, DNA and protein damage, motility changes, biofilm formation, and extracellular polysaccharide production. In addition, gene expression analysis by RT-qPCR showed its significant inhibition on the two-component system (TCS) and quorum sensing system (QS). The results indicated that FIT-β-CD exhibited efficient, sensitive photodynamic, and antibacterial properties against <em>E.coli</em> O157:H7, making it a novel photosensitizer. These findings imply potential new applications of fisetin in the field of food antimicrobial agents.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 145-152"},"PeriodicalIF":3.9,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145708122","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}
Pub Date : 2026-02-01Epub Date: 2025-12-16DOI: 10.1016/j.jbiotec.2025.12.011
Linhai Xie , Shuo Tian , Zhehao Jin , Tiantian Zou , Mingxin Chang , Hongting Tang , Tao Yu , Zhoukang Zhuang
As an emerging alternative protein, Single cell protein (SCP) has gained considerable traction in livestock feed applications. Furthermore, it presents substantial potential for replacing traditional dietary proteins for human consumption. Yet current research remains focused on strain screening with limited advances in cellular metabolic engineering. Due to the complexity of protein synthesis and processing, enhancing the SCP production efficiency in microbial chassis remains a major challenge. Here, we present a metabolic engineering approach to systematic engineer the cell wall of Saccharomyces cerevisiae for improving SCP production. In this study, we constructed a strain by gene modifications of genes related to cell wall biogenesis and associated signaling pathways, achieving a 32.6 % increase in cellular protein content. Additionally, comparative transcriptomics analysis uncovered the regulatory mechanism whereby remodeling of carbon and nitrogen metabolism governs cellular protein biosynthesis, providing new insight for the rational design of microbial cell factories with enhanced protein content.
{"title":"Systematic engineering of cell wall for improving single cell protein (SCP) production","authors":"Linhai Xie , Shuo Tian , Zhehao Jin , Tiantian Zou , Mingxin Chang , Hongting Tang , Tao Yu , Zhoukang Zhuang","doi":"10.1016/j.jbiotec.2025.12.011","DOIUrl":"10.1016/j.jbiotec.2025.12.011","url":null,"abstract":"<div><div>As an emerging alternative protein, Single cell protein (SCP) has gained considerable traction in livestock feed applications. Furthermore, it presents substantial potential for replacing traditional dietary proteins for human consumption. Yet current research remains focused on strain screening with limited advances in cellular metabolic engineering. Due to the complexity of protein synthesis and processing, enhancing the SCP production efficiency in microbial chassis remains a major challenge. Here, we present a metabolic engineering approach to systematic engineer the cell wall of <em>Saccharomyces cerevisiae</em> for improving SCP production. In this study, we constructed a strain by gene modifications of genes related to cell wall biogenesis and associated signaling pathways, achieving a 32.6 % increase in cellular protein content. Additionally, comparative transcriptomics analysis uncovered the regulatory mechanism whereby remodeling of carbon and nitrogen metabolism governs cellular protein biosynthesis, providing new insight for the rational design of microbial cell factories with enhanced protein content.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 173-183"},"PeriodicalIF":3.9,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145781399","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}
Pub Date : 2026-02-01Epub Date: 2025-11-21DOI: 10.1016/j.jbiotec.2025.11.013
E.M. Abou Hussein , Rasha Mohammad Fathy
A New glass system with a composition of; 50.3 B2O3+ 21 CaO + 8 MgO + 8 K2O + 8 Na2O + 4 P2O5 + 0.7 Nano Ag2O, was prepared by the traditional melting technique. Some of its structural, optical, and physical characteristics were investigated regarding the effect of gamma radiation. The XRD pattern revealed the amorphous nature of the fabricated glass sample. UV–visible absorption spectra revealed an obvious peak before 400 nm due to the absorption of Ag2 + ions, and a photo-reduction reaction to Ag0 was expected due to the impact of gamma irradiation. FTIR absorption spectra displayed structural bands in the region 800–1600 cm−1 recognized to the specific vibrations of BO3, BO4 and PO2. Results displayed optical and structural stability against gamma irradiation doses of 25 and 50 kGy, and revealed a positive effect of irradiation to enhance the antifungal efficiency of the glass or in sterilization processes before usage. The antifungal activity of borate bioglass doped with silver nanoparticles (BG/Ag) against Candida albicans and Candida tropicalis was assessed. BG/Ag was mostly effective against C. albicans. Furthermore, at a dosage of 40.0 mg/mL, BG/Ag considerably reduced biofilm formation of C. albicans and C. tropicalis (90.79 and 75.07 %). At 40 mg/mL, BG/Ag significantly reduced Candida viability, colony counts, and phospholipase activity. BG/Ag can be successfully applied as an attractive biomaterial for the medication of serious fungal infections. The novelty in the study is the synthesis of borate bioactive glass containing nanosilver- and the use of gamma irradiation to improve its biological properties.
{"title":"Optical, structural, and antifungal properties of nanosilver borate bioactive glass synthesized using gamma rays on the survival of Candida albicans and Candida tropicalis","authors":"E.M. Abou Hussein , Rasha Mohammad Fathy","doi":"10.1016/j.jbiotec.2025.11.013","DOIUrl":"10.1016/j.jbiotec.2025.11.013","url":null,"abstract":"<div><div>A New glass system with a composition of; 50.3 B<sub>2</sub>O<sub>3</sub>+ 21 CaO + 8 MgO + 8 K<sub>2</sub>O + 8 Na<sub>2</sub>O + 4 P<sub>2</sub>O<sub>5</sub> + 0.7 Nano Ag<sub>2</sub>O, was prepared by the traditional melting technique. Some of its structural, optical, and physical characteristics were investigated regarding the effect of gamma radiation. The XRD pattern revealed the amorphous nature of the fabricated glass sample. UV–visible absorption spectra revealed an obvious peak before 400 nm due to the absorption of Ag<sup>2 +</sup> ions, and a photo-reduction reaction to Ag<sup>0</sup> was expected due to the impact of gamma irradiation. FTIR absorption spectra displayed structural bands in the region 800–1600 cm<sup>−1</sup> recognized to the specific vibrations of BO<sub>3</sub>, BO<sub>4</sub> and PO<sub>2</sub>. Results displayed optical and structural stability against gamma irradiation doses of 25 and 50 kGy, and revealed a positive effect of irradiation to enhance the antifungal efficiency of the glass or in sterilization processes before usage. The antifungal activity of borate bioglass doped with silver nanoparticles (BG/Ag) against <em>Candida albicans</em> and <em>Candida tropicalis</em> was assessed. BG/Ag was mostly effective against <em>C. albicans</em>. Furthermore, at a dosage of 40.0 mg/mL, BG/Ag considerably reduced biofilm formation of <em>C. albicans</em> and <em>C. tropicalis</em> (90.79 and 75.07 %). At 40 mg/mL, BG/Ag significantly reduced <em>Candida</em> viability, colony counts, and phospholipase activity. BG/Ag can be successfully applied as an attractive biomaterial for the medication of serious fungal infections. The novelty in the study is the synthesis of borate bioactive glass containing nanosilver- and the use of gamma irradiation to improve its biological properties.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 33-44"},"PeriodicalIF":3.9,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145587571","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}
Pub Date : 2026-02-01Epub Date: 2025-12-01DOI: 10.1016/j.jbiotec.2025.11.020
Suhye Choi , Seung Hun Lee , Yunyeong Kim , Woo Young Jeon , Jung oh Ahn , Shashi Kant Bhatia , Kwon-Young Choi , Jeong Chan Joo , Yung-Hun Yang
Alkanediols are industrial compounds with diverse applications. However, the biological production of C5 diols is limited by the low substrate specificity and efficiency of carboxylic acid reductases (CARs) toward short to medium-chain substrates. We developed a whole-cell biotransformation system for efficient 1,4-pentanediol (1,4-PDO) production, aimed at developing a platform pathway for diol biosynthesis. By screening 10 CARs, we identified MsmCAR from Mycolicibacterium smegmatis MC2 155 as the most effective for C5 diol biosynthesis, with high specificity toward 4-hydroxyvaleric acid (4-HV). We constructed a whole-cell catalyst expressing MsmCAR and enhanced its performance by introducing ppk2b from Corynebacterium glutamicum ATCC 13032 and chnD from Acinetobacter sp., which encode proteins involved in cofactor regeneration. Systematic optimization of the reaction conditions including buffers, cofactors, metal ions, and cultivation parameters led to a maximum titer of 78.10 mM 1,4-PDO, with a productivity of 1.86 mM/h, when using 4-HV as a substrate. To expand the substrate scope to levulinic acid, 3-hydroxybutyrate dehydrogenase and formate dehydrogenase biosynthetic genes were introduced. This system yielded 22.10 mM 1,4-PDO within 10 h. This work highlights the potential of MsmCAR as a versatile biocatalyst and provides a sustainable strategy for producing short- to medium-chain diols from biomass-derived feedstocks.
{"title":"High specificity of MsmCAR toward 4-hydroxyvaleric acid enables efficient 1,4-pentanediol production from biomass-derived levulinic acid","authors":"Suhye Choi , Seung Hun Lee , Yunyeong Kim , Woo Young Jeon , Jung oh Ahn , Shashi Kant Bhatia , Kwon-Young Choi , Jeong Chan Joo , Yung-Hun Yang","doi":"10.1016/j.jbiotec.2025.11.020","DOIUrl":"10.1016/j.jbiotec.2025.11.020","url":null,"abstract":"<div><div>Alkanediols are industrial compounds with diverse applications. However, the biological production of C5 diols is limited by the low substrate specificity and efficiency of carboxylic acid reductases (CARs) toward short to medium-chain substrates. We developed a whole-cell biotransformation system for efficient 1,4-pentanediol (1,4-PDO) production, aimed at developing a platform pathway for diol biosynthesis. By screening 10 CARs, we identified <em>Msm</em>CAR from <em>Mycolicibacterium smegmatis</em> MC2 155 as the most effective for C5 diol biosynthesis, with high specificity toward 4-hydroxyvaleric acid (4-HV). We constructed a whole-cell catalyst expressing <em>Msm</em>CAR and enhanced its performance by introducing <em>ppk2b</em> from <em>Corynebacterium glutamicum</em> ATCC 13032 and <em>chnD</em> from <em>Acinetobacter</em> sp., which encode proteins involved in cofactor regeneration. Systematic optimization of the reaction conditions including buffers, cofactors, metal ions, and cultivation parameters led to a maximum titer of 78.10 mM 1,4-PDO, with a productivity of 1.86 mM/h, when using 4-HV as a substrate. To expand the substrate scope to levulinic acid, 3-hydroxybutyrate dehydrogenase and formate dehydrogenase biosynthetic genes were introduced. This system yielded 22.10 mM 1,4-PDO within 10 h. This work highlights the potential of <em>Msm</em>CAR as a versatile biocatalyst and provides a sustainable strategy for producing short- to medium-chain diols from biomass-derived feedstocks.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 115-124"},"PeriodicalIF":3.9,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145668412","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}
Pub Date : 2026-02-01Epub Date: 2025-12-16DOI: 10.1016/j.jbiotec.2025.12.012
Bochun Hu , Xiaoqi Xi , Qianjin Hou , Jiaxin Ou , Xiaoxuan Shen , Zining Wang , Jiahui Zhao , Cunduo Tang , Jihong Huang
To eliminate the requirement for pyruvate supplementation and to avoid lactate accumulation, a multi-enzyme cascade was systematically restructured by replacing lactate dehydrogenase (LDH) with water-forming NADH oxidase (EfNOx). In this revised system, RpEHL360V catalyzed the enantioconvergent hydrolysis of rac-4-chlorostyrene oxide (rac-1a) to (R)-1-(4-chlorophenyl)-1,2-ethanediol (1b), followed by stereoselective oxidation of (R)-1b to 4’-chloro-2-hydroxyacetophenone (1c) by BsBDHAI49L/V266L/G292A. Concurrently, EfNOx reduced molecular oxygen to water, regenerating NAD+ and sustaining redox balance. To further enhance intracellular cofactor turnover, BsBDHAI49L/V266L/G292A and EfNOx were co-expressed in Escherichia coli, generating the recombinant strain E. coli/Cbn. Through single-factor and L9(33) orthogonal array optimization, the one-pot in vivo cascade achieved maximal efficiency with lyophilized whole cells of E. coli/rpehL360V (5 mg DCW/mL) and E. coli/Cbn (35 mg DCW/mL) in sodium phosphate buffer (10 mM, pH 7.0) containing 5 mM NAD+ and 5 % (v/v) Tween-20 at 25 °C for 8 h under controlled agitation and aeration. Under these conditions, 25 mM rac-1a was converted to 1c with a yield of 92.1 % and a space-time yield (STY) of 2.9 mmol/L/h, representing a 3.7-fold increase relative to the original LDH-based system. The applicability of the cascade was further demonstrated using additional substrates (rac-2a to rac-7a), yielding the corresponding α-hydroxyketones at 39.1–94.5 % with STYs of 1.2–3.0 mmol/L/h. These findings establish BDH–NOx co-expression as a robust strategy for cofactor self-sufficiency and provide a scalable framework for the efficient in vivo synthesis of structurally diverse α-hydroxyketones.
{"title":"Engineered BDH–NOx co-expression in Escherichia coli enables highly efficient in vivo cascade catalysis for the transformation of racemic epoxides to α-hydroxyketones","authors":"Bochun Hu , Xiaoqi Xi , Qianjin Hou , Jiaxin Ou , Xiaoxuan Shen , Zining Wang , Jiahui Zhao , Cunduo Tang , Jihong Huang","doi":"10.1016/j.jbiotec.2025.12.012","DOIUrl":"10.1016/j.jbiotec.2025.12.012","url":null,"abstract":"<div><div>To eliminate the requirement for pyruvate supplementation and to avoid lactate accumulation, a multi-enzyme cascade was systematically restructured by replacing lactate dehydrogenase (LDH) with water-forming NADH oxidase (<em>Ef</em>NOx). In this revised system, <em>Rp</em>EH<sup>L360V</sup> catalyzed the enantioconvergent hydrolysis of <em>rac</em>-4-chlorostyrene oxide (<em>rac</em>-<strong>1a</strong>) to (<em>R</em>)-1-(4-chlorophenyl)-1,2-ethanediol (<strong>1b</strong>), followed by stereoselective oxidation of (<em>R</em>)-<strong>1b</strong> to 4’-chloro-2-hydroxyacetophenone (<strong>1c</strong>) by <em>Bs</em>BDHA<sup>I49L/V266L/G292A</sup>. Concurrently, <em>Ef</em>NOx reduced molecular oxygen to water, regenerating NAD<sup>+</sup> and sustaining redox balance. To further enhance intracellular cofactor turnover, <em>Bs</em>BDHA<sup>I49L/V266L/G292A</sup> and <em>Ef</em>NOx were co-expressed in <em>Escherichia coli</em>, generating the recombinant strain <em>E. coli</em>/C<em>bn</em>. Through single-factor and L9(3<sup>3</sup>) orthogonal array optimization, the one-pot <em>in vivo</em> cascade achieved maximal efficiency with lyophilized whole cells of <em>E. coli/rpeh</em><sup>L360V</sup> (5 mg DCW/mL) and <em>E. coli</em>/C<em>bn</em> (35 mg DCW/mL) in sodium phosphate buffer (10 mM, pH 7.0) containing 5 mM NAD<sup>+</sup> and 5 % (v/v) Tween-20 at 25 °C for 8 h under controlled agitation and aeration. Under these conditions, 25 mM <em>rac</em>-<strong>1a</strong> was converted to <strong>1c</strong> with a yield of 92.1 % and a space-time yield (STY) of 2.9 mmol/L/h, representing a 3.7-fold increase relative to the original LDH-based system. The applicability of the cascade was further demonstrated using additional substrates (<em>rac</em>-<strong>2a</strong> to <em>rac</em>-<strong>7a</strong>), yielding the corresponding α-hydroxyketones at 39.1–94.5 % with STYs of 1.2–3.0 mmol/L/h. These findings establish BDH–NOx co-expression as a robust strategy for cofactor self-sufficiency and provide a scalable framework for the efficient <em>in vivo</em> synthesis of structurally diverse α-hydroxyketones.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 184-193"},"PeriodicalIF":3.9,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145781375","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}
Pub Date : 2026-02-01Epub Date: 2025-12-05DOI: 10.1016/j.jbiotec.2025.12.002
Jiao Liu, Chaolong Qu, Bin Zhang, Jie Bao
Trace amounts of D-lactic acid derived from lignocellulose feedstock and nitrogen additives frequently reduce the chiral purity of L-lactic acid below the polymerization standard during biorefinery fermentations. The spontaneous D-lactic acid degradation catalyzed by the highly substrate specific D-lactate oxidase in lactic acid bacterium provides an ideal solution. This study used a D-lactate oxidase GOX2071 with relatively high activity for construction of D-lactic acid oxidation pathway in the L-lactic acid producing strain Pediococcus acidilactici ZY271. The engineered P. acidilactici LJ2071 strain demonstrated an active D-lactic acid oxidation performance while the high L-lactic acid production property was well maintained. A L-lactic acid chiral purity of 99.63 % was obtained using the D-lactic acid containing wheat straw as feedstock and 40 % of corn steep liquor (CSL) as nitrogen additive to alternate expensive yeast extract. This study provided a practical method for upgrading the chirality of cellulosic L-lactic acid using lignocellulose feedstock and cheap nitrogen additives.
{"title":"Increasing the chiral purity of cellulosic L-lactic acid by D-Lactate oxidase-catalyzed oxidation in engineered lactic acid bacterium Pediococcus acidilactici","authors":"Jiao Liu, Chaolong Qu, Bin Zhang, Jie Bao","doi":"10.1016/j.jbiotec.2025.12.002","DOIUrl":"10.1016/j.jbiotec.2025.12.002","url":null,"abstract":"<div><div>Trace amounts of <span>D</span>-lactic acid derived from lignocellulose feedstock and nitrogen additives frequently reduce the chiral purity of <span>L</span>-lactic acid below the polymerization standard during biorefinery fermentations. The spontaneous <span>D</span>-lactic acid degradation catalyzed by the highly substrate specific <span>D</span>-lactate oxidase in lactic acid bacterium provides an ideal solution. This study used a <span>D</span>-lactate oxidase GOX2071 with relatively high activity for construction of <span>D</span>-lactic acid oxidation pathway in the <span>L</span>-lactic acid producing strain <em>Pediococcus acidilactici</em> ZY271. The engineered <em>P</em>. <em>acidilactici</em> LJ2071 strain demonstrated an active <span>D</span>-lactic acid oxidation performance while the high <span>L</span>-lactic acid production property was well maintained. A <span>L</span>-lactic acid chiral purity of 99.63 % was obtained using the <span>D</span>-lactic acid containing wheat straw as feedstock and 40 % of corn steep liquor (CSL) as nitrogen additive to alternate expensive yeast extract. This study provided a practical method for upgrading the chirality of cellulosic <span>L</span>-lactic acid using lignocellulose feedstock and cheap nitrogen additives.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 137-144"},"PeriodicalIF":3.9,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145701077","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}
Pub Date : 2026-02-01Epub Date: 2025-12-15DOI: 10.1016/j.jbiotec.2025.12.006
Johanna Pechan , Kristina Engström , Nora Mac Key , Lina Sörvik , Véronique Chotteau
Human Mesenchymal Stromal Cells (hMSCs) are a safe option for allogeneic cell therapy across various diseases, but their manufacturing process requires improvement to broaden accessibility. In this study, a state-of-the-art planar multi-vessel process was transferred to a stirred tank bioreactor using microcarriers to support the growth of adherent cells. The frequent medium exchange strategy from planar culture guided the design of the bioreactor process. However, complete medium changes in the bioreactor resulted in limited cell expansion and higher glucose consumption compared to planar culture. To enhance expansion, a 0.1 L perfusion bioreactor was tested, enabling continuous medium exchange. Three perfusion approaches were evaluated: (1) maintaining a target cell-specific glucose consumption rate, (2) varying the perfusion rate, and (3) applying a cell-specific perfusion rate. Implementing targeted glucose feeding (TAFE) reduced lactate production, while increasing perfusion rates improved cell density. The highest expansion was achieved using a cell-specific perfusion rate of 5 nL cell−1 day−1 combined with a target glucose consumption rate (qglc) of 15 pmol cell−1 day−1, resulting in a 5.4-fold higher expansion factor than daily medium changes in stirred tank bioreactors. This optimized process represents a key advancement toward producing clinically relevant quantities of hMSCs.
{"title":"Cell specific nutrient feeding in perfusion mode enhances hMSC growth in stirred tank bioreactor process","authors":"Johanna Pechan , Kristina Engström , Nora Mac Key , Lina Sörvik , Véronique Chotteau","doi":"10.1016/j.jbiotec.2025.12.006","DOIUrl":"10.1016/j.jbiotec.2025.12.006","url":null,"abstract":"<div><div>Human Mesenchymal Stromal Cells (hMSCs) are a safe option for allogeneic cell therapy across various diseases, but their manufacturing process requires improvement to broaden accessibility. In this study, a state-of-the-art planar multi-vessel process was transferred to a stirred tank bioreactor using microcarriers to support the growth of adherent cells. The frequent medium exchange strategy from planar culture guided the design of the bioreactor process. However, complete medium changes in the bioreactor resulted in limited cell expansion and higher glucose consumption compared to planar culture. To enhance expansion, a 0.1 L perfusion bioreactor was tested, enabling continuous medium exchange. Three perfusion approaches were evaluated: (1) maintaining a target cell-specific glucose consumption rate, (2) varying the perfusion rate, and (3) applying a cell-specific perfusion rate. Implementing targeted glucose feeding (TAFE) reduced lactate production, while increasing perfusion rates improved cell density. The highest expansion was achieved using a cell-specific perfusion rate of 5 nL cell<sup>−1</sup> day<sup>−1</sup> combined with a target glucose consumption rate (q<sub>glc</sub>) of 15 pmol cell<sup>−1</sup> day<sup>−1</sup>, resulting in a 5.4-fold higher expansion factor than daily medium changes in stirred tank bioreactors. This optimized process represents a key advancement toward producing clinically relevant quantities of hMSCs.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 194-206"},"PeriodicalIF":3.9,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145774742","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}
The rising incidence of Helicobacter pylori-associated infections and colorectal cancer necessitates multifunctional therapeutics targeting both microbial and tumor pathologies. However, antimicrobial peptides (AMPs) like human β-defensin 3 (hBD-3) faces challenges such as proteolytic degradation and poor bioavailability. To address these limitations, we developed a novel hybrid nanozyme by immobilizing hBD-3 onto a guanosine monophosphate-coordinated iron framework (Fe/GMP), known for its biocompatibility and intrinsic peroxidase-like activity.
Methods
The hBD-3@Fe/GMP nanozyme was synthesized and characterized for its physicochemical and catalytic properties. Its antibacterial activity against H. pylori was assessed via agar diffusion, biofilm inhibition, and virulence gene expression. Anti-colon cancer potential was evaluated in HT-29 cells using assays for cytotoxicity, ROS generation, apoptosis, migration inhibition, and redox biomarkers. in vivo antitumor efficacy was studied using a colon carcinoma xenograft mice model.
Results
hBD-3@Fe/GMP showed enhanced peroxidase-like activity with elevated ROS production than Fe/GMP alone. It significantly inhibited H. pylori growth, biofilm formation, and virulence gene expression (babA, alpA, alpB, flaA, flaB). In HT-29 cells, it induced cytotoxicity, suppressed antioxidant enzymes, and triggered apoptosis via p53 and Bax upregulation and Bcl-2 downregulation. In tumor xenograft model, it reduced tumor volume and weight.
Conclusion
This study reports, for the first time, a hybrid nanozyme that synergistically integrates the antimicrobial potency of hBD-3 with the catalytic redox functionality of Fe/GMP, offering dual action against H. pylori infection and colorectal cancer. The multifunctional therapeutic potential and biocompatibility of hBD-3@Fe/GMP mark a novel paradigm in the development of next-generation nanozymes for combinatorial infectious and oncological interventions.
{"title":"Human beta-defensin 3-functionalized Fe/GMP nanozyme for multifunctional antimicrobial and anticancer activity against Helicobacter pylori-associated gastrointestinal cancer","authors":"Yoganathan Kamaraj , Veenayohini Kumaresan , Jinhao Hu , Daochen Zhu","doi":"10.1016/j.jbiotec.2025.11.018","DOIUrl":"10.1016/j.jbiotec.2025.11.018","url":null,"abstract":"<div><h3>Background</h3><div>The rising incidence of <em>Helicobacter pylori</em>-associated infections and colorectal cancer necessitates multifunctional therapeutics targeting both microbial and tumor pathologies. However, antimicrobial peptides (AMPs) like human β-defensin 3 (hBD-3) faces challenges such as proteolytic degradation and poor bioavailability. To address these limitations, we developed a novel hybrid nanozyme by immobilizing hBD-3 onto a guanosine monophosphate-coordinated iron framework (Fe/GMP), known for its biocompatibility and intrinsic peroxidase-like activity.</div></div><div><h3>Methods</h3><div>The hBD-3@Fe/GMP nanozyme was synthesized and characterized for its physicochemical and catalytic properties. Its antibacterial activity against <em>H. pylori</em> was assessed via agar diffusion, biofilm inhibition, and virulence gene expression. Anti-colon cancer potential was evaluated in HT-29 cells using assays for cytotoxicity, ROS generation, apoptosis, migration inhibition, and redox biomarkers. <em>in vivo</em> antitumor efficacy was studied using a colon carcinoma xenograft mice model.</div></div><div><h3>Results</h3><div>hBD-3@Fe/GMP showed enhanced peroxidase-like activity with elevated ROS production than Fe/GMP alone. It significantly inhibited <em>H. pylori</em> growth, biofilm formation, and virulence gene expression (<em>babA, alpA, alpB, flaA, flaB</em>). In HT-29 cells, it induced cytotoxicity, suppressed antioxidant enzymes, and triggered apoptosis via p53 and Bax upregulation and Bcl-2 downregulation. In tumor xenograft model, it reduced tumor volume and weight.</div></div><div><h3>Conclusion</h3><div>This study reports, for the first time, a hybrid nanozyme that synergistically integrates the antimicrobial potency of hBD-3 with the catalytic redox functionality of Fe/GMP, offering dual action against <em>H. pylori</em> infection and colorectal cancer. The multifunctional therapeutic potential and biocompatibility of hBD-3@Fe/GMP mark a novel paradigm in the development of next-generation nanozymes for combinatorial infectious and oncological interventions.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 67-83"},"PeriodicalIF":3.9,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145622128","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}
Pub Date : 2026-02-01Epub Date: 2025-12-12DOI: 10.1016/j.jbiotec.2025.12.005
Chunxiao Yan , Shuai Zhang , Zhihao Huang , Fei Du , Qian Xu , Wang Ma , Guang Yang , Xiaoman Sun
As an excellent host for docosahexaenoic acid (DHA) biosynthesis, Schizochytrium sp. still faces inherent limitations in DHA yield due to oxidative damage induced by reactive oxygen species (ROS). To effectively reduce the ROS levels, the optimal combination of mixed antioxidants (52.95 mg L−1 phytic acid, 32.00 mg L−1 licorice extract, 36.28 mg L−1 ascorbic acid palmitate) was selected by single factor experiment combined with response surface optimization. The DHA titer increased from 24.0 g L−1 to 33.5 g L−1. However, exogenous addition of antioxidants results in increased the cost of DHA production. Enhancing the intracellular antioxidant capacity through gene engineering has become another feasible strategy. Transcriptome analysis indicated that the transcriptional levels of antioxidant-related genes SOD1 (superoxide dismutase), APX5 (ascorbic acid peroxidase), and GST3 (glutathione S-transferase) were significantly upregulated during DHA biosynthesis. Then, the engineered strain SOD1-APX5-GST3–2 was constructed by overexpressing the three genes, and the DHA and lipid titers were 33.1 g L−1 and 64.1 g L−1, respectively. Scale-up in a 5 L bioreactor obtained 32.5 g L−1 DHA and 63.5 g L−1 lipid, and the ROS level decreased by 30.5 %, demonstrating engineered strain's stability and laying the foundation for industrial-scale production. This study verified that reducing ROS levels promoted DHA biosynthesis in Schizochytrium sp. from the two dimensions of exogenous antioxidants and gene engineering, which provided novel insights for industrial lipid production.
{"title":"Enhancing oxidative stress defense to improve docosahexaenoic acid production in Schizochytrium sp. HX-308","authors":"Chunxiao Yan , Shuai Zhang , Zhihao Huang , Fei Du , Qian Xu , Wang Ma , Guang Yang , Xiaoman Sun","doi":"10.1016/j.jbiotec.2025.12.005","DOIUrl":"10.1016/j.jbiotec.2025.12.005","url":null,"abstract":"<div><div>As an excellent host for docosahexaenoic acid (DHA) biosynthesis, <em>Schizochytrium</em> sp. still faces inherent limitations in DHA yield due to oxidative damage induced by reactive oxygen species (ROS). To effectively reduce the ROS levels, the optimal combination of mixed antioxidants (52.95 mg L<sup>−1</sup> phytic acid, 32.00 mg L<sup>−1</sup> licorice extract, 36.28 mg L<sup>−1</sup> ascorbic acid palmitate) was selected by single factor experiment combined with response surface optimization. The DHA titer increased from 24.0 g L<sup>−1</sup> to 33.5 g L<sup>−1</sup>. However, exogenous addition of antioxidants results in increased the cost of DHA production. Enhancing the intracellular antioxidant capacity through gene engineering has become another feasible strategy. Transcriptome analysis indicated that the transcriptional levels of antioxidant-related genes <em>SOD1</em> (superoxide dismutase), <em>APX5</em> (ascorbic acid peroxidase), and <em>GST3</em> (glutathione S-transferase) were significantly upregulated during DHA biosynthesis. Then, the engineered strain SOD1-APX5-GST3–2 was constructed by overexpressing the three genes, and the DHA and lipid titers were 33.1 g L<sup>−1</sup> and 64.1 g L<sup>−1</sup>, respectively. Scale-up in a 5 L bioreactor obtained 32.5 g L<sup>−1</sup> DHA and 63.5 g L<sup>−1</sup> lipid, and the ROS level decreased by 30.5 %, demonstrating engineered strain's stability and laying the foundation for industrial-scale production. This study verified that reducing ROS levels promoted DHA biosynthesis in <em>Schizochytrium</em> sp. from the two dimensions of exogenous antioxidants and gene engineering, which provided novel insights for industrial lipid production.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 162-172"},"PeriodicalIF":3.9,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145756688","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}
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