Pub 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":"2025-12-05","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 : 2025-12-04DOI: 10.1016/j.jbiotec.2025.12.001
Jiaxin Li , Weiyang Zhao , Yunlei Fu , Ranxuan Li , Jin Liu , Shufang Yang , Jie Zheng , Haijin Mou , Han Sun
Large-scale production of microalgal lutein faces challenges due to limited biomass and lutein content. In this work, we developed an exponential fed-batch model to achieve heterotrophic high-density growth of Chlorella protothecoides CS-41. Then, indole-3-acetic acid (IAA) and high nitrogen were combined to promote lutein accumulation in situ. Fed-batch culture enhanced the TCA cycle of C. protothecoides CS-41 to benefit cell growth while increasing carotenoid flux to facilitate lutein accumulation through redirecting carbon flux towards lutein biosynthesis, improved by IAA. Also, high nitrogen increased lutein content through modulation of CMS (carotenoid methyltransferase synthase), CRTISO (carotenoid isomerase), LCYB (lycopene β-cyclase), ZEP (zeaxanthin epoxidase), and VDE (violaxanthin de-epoxidase). Finally, under heterotrophic conditions, combining 200 mM IAA with 3.6 g/L urea improved pyruvate and 3-phosphoglyceric acid and then boosted lutein content and productivity to 3.27 mg/g and 20.38 mg/L/d, respectively, by increases of 58 % and 56.7 %. Addressing issues including density dilution, container transfer, and in-situ lutein accumulation under heterotrophic high-density conditions offers a cost-effective solution.
由于生物量和叶黄素含量有限，微藻叶黄素的大规模生产面临挑战。本研究建立了小球藻CS-41异养高密度生长的指数补料批量模型。然后，吲哚-3-乙酸（IAA）和高氮配合施用，促进叶黄素原位积累。补料批培养增强了C. protothecoides CS-41的TCA循环，有利于细胞生长，同时增加类胡萝卜素通量，通过将碳通量转向叶黄素的生物合成，促进叶黄素的积累，IAA改善了这一过程。高氮通过调节类胡萝卜素甲基转移酶合成酶（CMS）、类胡萝卜素异构酶（CRTISO）、番茄红素β环化酶（LCYB）、玉米黄质环氧化酶（ZEP）和紫黄质去环氧化酶（VDE）提高叶黄素含量。最后，在异养条件下，200 mM IAA与3.6 g/L尿素配用可改善丙酮酸和3-磷酸甘油酸，叶黄素含量和产量分别提高了3.27 mg/g和20.38 mg/L/d，分别提高了58 %和56.7% %。在异养高密度条件下，解决密度稀释、容器转移和原位叶黄素积累等问题提供了具有成本效益的解决方案。
{"title":"Enhancing heterotrophic lutein production in Chlorella protothecoides through combined phytohormone and nitrogen strategies","authors":"Jiaxin Li , Weiyang Zhao , Yunlei Fu , Ranxuan Li , Jin Liu , Shufang Yang , Jie Zheng , Haijin Mou , Han Sun","doi":"10.1016/j.jbiotec.2025.12.001","DOIUrl":"10.1016/j.jbiotec.2025.12.001","url":null,"abstract":"<div><div>Large-scale production of microalgal lutein faces challenges due to limited biomass and lutein content. In this work, we developed an exponential fed-batch model to achieve heterotrophic high-density growth of <em>Chlorella protothecoides</em> CS-41. Then, indole-3-acetic acid (IAA) and high nitrogen were combined to promote lutein accumulation <em>in situ</em>. Fed-batch culture enhanced the TCA cycle of <em>C. protothecoides</em> CS-41 to benefit cell growth while increasing carotenoid flux to facilitate lutein accumulation through redirecting carbon flux towards lutein biosynthesis, improved by IAA. Also, high nitrogen increased lutein content through modulation of <em>CMS</em> (carotenoid methyltransferase synthase), <em>CRTISO</em> (carotenoid isomerase), <em>LCYB</em> (lycopene β-cyclase), <em>ZEP</em> (zeaxanthin epoxidase), and <em>VDE</em> (violaxanthin de-epoxidase). Finally, under heterotrophic conditions, combining 200 mM IAA with 3.6 g/L urea improved pyruvate and 3-phosphoglyceric acid and then boosted lutein content and productivity to 3.27 mg/g and 20.38 mg/L/d, respectively, by increases of 58 % and 56.7 %. Addressing issues including density dilution, container transfer, and <em>in-situ</em> lutein accumulation under heterotrophic high-density conditions offers a cost-effective solution.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 125-136"},"PeriodicalIF":3.9,"publicationDate":"2025-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145681983","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 : 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":"2025-12-01","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}
Enzymes are widely used in various industries due to their eco-friendly nature, high selectivity, and ability to operate under mild conditions. However, large-scale applications are often hindered by challenges such as limited stability, high costs, and difficulties in recyclability. Enzyme immobilization presents a promising strategy to overcome these limitations. Microbial collagenases, particularly those from Clostridium histolyticum, play a crucial role in industrial and biomedical applications. In silico analyses effectively predict enzyme immobilization outcomes, minimizing experimental time and cost. This study employed enzyme surface analysis, a bioinformatics approach, to determine the surface charge type of the support for the physical immobilization of C. histolyticum collagenases. The analysis identified the most suitable regions on the enzyme surfaces for attachment—positively charged and distant from critical sites—ensuring minimal disruption to activity. Copper oxide (CuO) nanoparticles (NPs) were synthesized using walnut green husk extract and subsequently characterized. The green-synthesized CuO NPs were monodisperse and spherical (∼40 nm) with a negatively charged surface, confirming their suitability as supports for enzyme immobilization. Employing the CuO NPs as supports resulted in an immobilization yield of 42.15 % and an activity yield of 146.2 %. Further optimization of immobilization conditions could improve IY. The high AY suggests that immobilization did not hinder enzyme function but may have enhanced enzyme-substrate affinity and increased local substrate concentration, thereby boosting enzyme efficiency.
{"title":"Experimental immobilization of Clostridium histolyticum collagenases on green-synthesized CuO nanoparticles supported by bioinformatics analysis","authors":"Farzaneh Barati , Fakhrisadat Hosseini , Parinaz Ghadam , Seyed Shahriar Arab","doi":"10.1016/j.jbiotec.2025.11.019","DOIUrl":"10.1016/j.jbiotec.2025.11.019","url":null,"abstract":"<div><div>Enzymes are widely used in various industries due to their eco-friendly nature, high selectivity, and ability to operate under mild conditions. However, large-scale applications are often hindered by challenges such as limited stability, high costs, and difficulties in recyclability. Enzyme immobilization presents a promising strategy to overcome these limitations. Microbial collagenases, particularly those from <em>Clostridium histolyticum</em>, play a crucial role in industrial and biomedical applications. <em>In silico</em> analyses effectively predict enzyme immobilization outcomes, minimizing experimental time and cost. This study employed enzyme surface analysis, a bioinformatics approach, to determine the surface charge type of the support for the physical immobilization of <em>C. histolyticum</em> collagenases. The analysis identified the most suitable regions on the enzyme surfaces for attachment—positively charged and distant from critical sites—ensuring minimal disruption to activity. Copper oxide (CuO) nanoparticles (NPs) were synthesized using walnut green husk extract and subsequently characterized. The green-synthesized CuO NPs were monodisperse and spherical (∼40 nm) with a negatively charged surface, confirming their suitability as supports for enzyme immobilization. Employing the CuO NPs as supports resulted in an immobilization yield of 42.15 % and an activity yield of 146.2 %. Further optimization of immobilization conditions could improve IY. The high AY suggests that immobilization did not hinder enzyme function but may have enhanced enzyme-substrate affinity and increased local substrate concentration, thereby boosting enzyme efficiency.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 105-114"},"PeriodicalIF":3.9,"publicationDate":"2025-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145654360","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 : 2025-11-27DOI: 10.1016/j.jbiotec.2025.11.014
Fubin Yin , Tianjing Lian , Haoying Qu , Dongan Cui , Xiaomeng Li , Yangyang Cai , Hongmin Dong
The technology of directional lactic acid production (DLAP) for animal manure is an innovative way of value-added utilization, but the influence of residual antibiotics in animal manure on the DLAP is still unclear, especially the degradation rate and degradation pathways of antibiotic with residues lactic acid bacteria (LAB). In this study, the most common residual antibiotic (Sulfachloropyridazine, SCP) and LAB (Lactobacillus bulgaricus, Lactobacillus acidophilus, Lactobacillus plantarum and Lactobacillus casei) were employed as the research objects, the SCP degradation, degradation pathways and antibiotic resistance genes (ARGs) with different LAB were investigated in pure culture. Results showed that LAB has a positive effect on the reduction in ARGs (sul1, sul2, sul3, sulA, intI1 and tnp A). The SCP degradation rates were 90.3 %, 85.1 %, 86.1 % and 84.0 % with the growth of Lactobacillus bulgaricus, Lactobacillus acidophilus, Lactobacillus plantarum and Lactobacillus casei at 50 mg/L, respectively. And the SCP degradation pathway is different for LAB, but the main decomposition functions, including S-N bond cleavage, CH3 release, SO2 release, nitration, hydroxylation, NH2 release and C-N bond cleavage, are similar. And most of intermediate products of SCP were lower toxicity compared to the parent compound. The findings proved the DLAP process as a guidance for a potential solution for reducing antibiotic and ARGs contamination.
{"title":"Directional lactic acid production could mitigate antibiotic and ARGs pollution","authors":"Fubin Yin , Tianjing Lian , Haoying Qu , Dongan Cui , Xiaomeng Li , Yangyang Cai , Hongmin Dong","doi":"10.1016/j.jbiotec.2025.11.014","DOIUrl":"10.1016/j.jbiotec.2025.11.014","url":null,"abstract":"<div><div>The technology of directional lactic acid production (DLAP) for animal manure is an innovative way of value-added utilization, but the influence of residual antibiotics in animal manure on the DLAP is still unclear, especially the degradation rate and degradation pathways of antibiotic with residues lactic acid bacteria (LAB). In this study, the most common residual antibiotic (Sulfachloropyridazine, SCP) and LAB (<em>Lactobacillus bulgaricus, Lactobacillus acidophilus, Lactobacillus plantarum and Lactobacillus casei</em>) were employed as the research objects, the SCP degradation, degradation pathways and antibiotic resistance genes (ARGs) with different LAB were investigated in pure culture. Results showed that LAB has a positive effect on the reduction in ARGs (sul1, sul2, sul3, sulA, intI1 and tnp A). The SCP degradation rates were 90.3 %, 85.1 %, 86.1 % and 84.0 % with the growth of <em>Lactobacillus bulgaricus</em>, <em>Lactobacillus acidophilus</em>, <em>Lactobacillus plantarum</em> and <em>Lactobacillus casei</em> at 50 mg/L, respectively. And the SCP degradation pathway is different for LAB, but the main decomposition functions, including S-N bond cleavage, CH<sub>3</sub> release, SO<sub>2</sub> release, nitration, hydroxylation, NH<sub>2</sub> release and C-N bond cleavage, are similar. And most of intermediate products of SCP were lower toxicity compared to the parent compound. The findings proved the DLAP process as a guidance for a potential solution for reducing antibiotic and ARGs contamination.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 96-104"},"PeriodicalIF":3.9,"publicationDate":"2025-11-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145622130","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}
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":"2025-11-27","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}
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":"2025-11-26","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 : 2025-11-25DOI: 10.1016/j.jbiotec.2025.11.016
Yajie Wang , Ruixue Ma , Jia Zhou , Zhongbiao Tan , Hongliang Li , Hao Shi , Yegui Zhang
Glutathione (GSH), a ubiquitous bioactive tripeptide, plays a central role in antioxidant defense, detoxification, and cellular metabolism. This review highlights advanced strategies to enhance GSH production, emphasizing strain improvement via traditional mutagenesis and genetic engineering. Optimization of microbial fermentation through carbon source selection, amino acid precursors, and ATP regeneration combined with metabolic engineering approaches, has significantly improved GSH yields in Saccharomyces cerevisiae and Escherichia coli. Clinically, GSH is used in managing liver diseases, oxidative stress, and immune dysfunction. Emerging applications, particularly in oncology, show promise. This review also compares production methods such as chemical synthesis, enzymatic catalysis, and microbial fermentation, highlighting microbial fermentation for its sustainability, cost-efficiency, and scalability. Future research should focus on refining detection methods, elucidating GSH’s therapeutic mechanisms, and broadening its applications across pharmaceuticals, nutrition, and cosmetics.
{"title":"Emerging strategies for enhanced glutathione biosynthesis and its biomedical applications","authors":"Yajie Wang , Ruixue Ma , Jia Zhou , Zhongbiao Tan , Hongliang Li , Hao Shi , Yegui Zhang","doi":"10.1016/j.jbiotec.2025.11.016","DOIUrl":"10.1016/j.jbiotec.2025.11.016","url":null,"abstract":"<div><div>Glutathione (GSH), a ubiquitous bioactive tripeptide, plays a central role in antioxidant defense, detoxification, and cellular metabolism. This review highlights advanced strategies to enhance GSH production, emphasizing strain improvement via traditional mutagenesis and genetic engineering. Optimization of microbial fermentation through carbon source selection, amino acid precursors, and ATP regeneration combined with metabolic engineering approaches, has significantly improved GSH yields in <em>Saccharomyces cerevisiae</em> and <em>Escherichia coli</em>. Clinically, GSH is used in managing liver diseases, oxidative stress, and immune dysfunction. Emerging applications, particularly in oncology, show promise. This review also compares production methods such as chemical synthesis, enzymatic catalysis, and microbial fermentation, highlighting microbial fermentation for its sustainability, cost-efficiency, and scalability. Future research should focus on refining detection methods, elucidating GSH’s therapeutic mechanisms, and broadening its applications across pharmaceuticals, nutrition, and cosmetics.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 45-56"},"PeriodicalIF":3.9,"publicationDate":"2025-11-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145622161","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 : 2025-11-25DOI: 10.1016/j.jbiotec.2025.11.015
Lifen He , Danling Tang , Fan Feng , Tao Hong , Mingjing Zheng , Xiping Du , Yanbing Zhu , Yuanfan Yang , Zedong Jiang , Fan He , Lijun Li , Zhipeng Li , Hui Ni , Qingbiao Li
Pichia pastoris is a prominent host for recombinant protein production, utilizing the methanol-inducible Alcohol Oxidase 1 (AOX1) promoter to express thousands of heterologous proteins. However, methanol induction, while driving high-level protein expression, simultaneously imposes a pronounced metabolic burden. This review highlights that a balanced and ample supply of amino acids is indispensable for efficient protein synthesis. The induction process leads to the accumulation of by-products, notably higher alcohols, which can negatively impact protein quality. The dynamic interplay between these two pathways is not fully understood. In addition, the lack of quantitative models for their competition over shared precursors hinders rational metabolic engineering and impedes the industrial exploitation of P. pastoris. Therefore, this review systematically dissects the link between methanol induction, amino acid metabolism, and higher alcohol synthesis. It specifically examines how methanol-triggered perturbations in amino acid metabolism propagate to modulate higher alcohol formation. Special attention is given to key metabolic nodes, such as pyruvate and α-ketoisovalerate, where carbon flux diverges between biosynthesis and by-product formation. This review provides significant implications for a deeper understanding of the metabolic processes in P. pastoris, offering comprehensive mechanistic insights and guidance for optimizing yeast fermentation and enhancing recombinant protein production.
{"title":"Methanol-induced metabolic disturbances in protein expression in Pichia pastoris: A review of the interaction between amino acid synthesis and higher alcohol pathways","authors":"Lifen He , Danling Tang , Fan Feng , Tao Hong , Mingjing Zheng , Xiping Du , Yanbing Zhu , Yuanfan Yang , Zedong Jiang , Fan He , Lijun Li , Zhipeng Li , Hui Ni , Qingbiao Li","doi":"10.1016/j.jbiotec.2025.11.015","DOIUrl":"10.1016/j.jbiotec.2025.11.015","url":null,"abstract":"<div><div><em>Pichia pastoris</em> is a prominent host for recombinant protein production, utilizing the methanol-inducible Alcohol Oxidase 1 (AOX1) promoter to express thousands of heterologous proteins. However, methanol induction, while driving high-level protein expression, simultaneously imposes a pronounced metabolic burden. This review highlights that a balanced and ample supply of amino acids is indispensable for efficient protein synthesis. The induction process leads to the accumulation of by-products, notably higher alcohols, which can negatively impact protein quality. The dynamic interplay between these two pathways is not fully understood. In addition, the lack of quantitative models for their competition over shared precursors hinders rational metabolic engineering and impedes the industrial exploitation of <em>P. pastoris</em>. Therefore, this review systematically dissects the link between methanol induction, amino acid metabolism, and higher alcohol synthesis. It specifically examines how methanol-triggered perturbations in amino acid metabolism propagate to modulate higher alcohol formation. Special attention is given to key metabolic nodes, such as pyruvate and α-ketoisovalerate, where carbon flux diverges between biosynthesis and by-product formation. This review provides significant implications for a deeper understanding of the metabolic processes in <em>P. pastoris</em>, offering comprehensive mechanistic insights and guidance for optimizing yeast fermentation and enhancing recombinant protein production.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 57-66"},"PeriodicalIF":3.9,"publicationDate":"2025-11-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145622160","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 : 2025-11-21DOI: 10.1016/j.jbiotec.2025.11.012
Lian Zhang , Xiaojuan Shen , Zhi Wang , Yuansong Peng , Jianping Yang , Yafan Cai , Shilei Wang , Jingliang Xu , Hanjie Ying
Chinese distillers grains (CDGs) are used as low-cost feed additives, because they have issues such as high crude fiber content (> 24 %), low crude protein content (< 17 %), and low digestibility (< 16 %). This study aimed to evaluate whether pre-secondary solid-state fermentation using functional microbiota can improve protein digestibility and reduce fiber content in CDGs. Results indicated that compared to CDGs, both true and crude protein content in solid-state fermented DCDGs (SSF-DCDGs) showed an upward trend, reaching increases of over 24 %. Compared to DCDGs, crude fiber content decreased; dry matter and protein digestibility increased by 29.54 % and 54.57 % in SSF-DCDGs. Amino acid content rose by 14.92 %, with glycine and alanine increased by 15.36 % and 21.25 % respectively—both being important aromatic flavor compounds. Therefore, the Pre-Sec-SSF-FM technical route can convert low-cost CDGs into a high-nutritional protein sources, offering a novel approach for protein raw material sourcing.
{"title":"Production of high-nutrient protein sources by pre-secondary solid-state fermentation of functional microbiota (Pre-Sec-SSF-FM) from dehulled chinese distillers grains","authors":"Lian Zhang , Xiaojuan Shen , Zhi Wang , Yuansong Peng , Jianping Yang , Yafan Cai , Shilei Wang , Jingliang Xu , Hanjie Ying","doi":"10.1016/j.jbiotec.2025.11.012","DOIUrl":"10.1016/j.jbiotec.2025.11.012","url":null,"abstract":"<div><div>Chinese distillers grains (CDGs) are used as low-cost feed additives, because they have issues such as high crude fiber content (> 24 %), low crude protein content (< 17 %), and low digestibility (< 16 %). This study aimed to evaluate whether pre-secondary solid-state fermentation using functional microbiota can improve protein digestibility and reduce fiber content in CDGs. Results indicated that compared to CDGs, both true and crude protein content in solid-state fermented DCDGs (SSF-DCDGs) showed an upward trend, reaching increases of over 24 %. Compared to DCDGs, crude fiber content decreased; dry matter and protein digestibility increased by 29.54 % and 54.57 % in SSF-DCDGs. Amino acid content rose by 14.92 %, with glycine and alanine increased by 15.36 % and 21.25 % respectively—both being important aromatic flavor compounds. Therefore, the Pre-Sec-SSF-FM technical route can convert low-cost CDGs into a high-nutritional protein sources, offering a novel approach for protein raw material sourcing.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 23-32"},"PeriodicalIF":3.9,"publicationDate":"2025-11-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145587491","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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