Herein, glucose oxidase was immobilized into bimetallic (Cu2+, Zn2+) hybrid magnetic nanoflowers (GOx-mcNFs) to perform as a more stable glucose biosensor and antibacterial material. Morphological characterizations of GOx-mcNFs verified the construction of nanoflowers through metal ions-GOx coordination. The encapsulation yield was obtained as 93.5 %, where the activity recovery was 650.3 %. Magnetic nanoparticles also enhanced the peroxidase-like activity of metal-phosphates and caused GOx-mcNFs to show excellent peroxidase-like properties with the apparent Km values of 0.19 mM and 0.267 mM for H2O2 and TMB, respectively. Thermodynamic studies revealed a 16.5 kJ mol−1 higher activation energy of denaturation for GOx-mcNFs over the free GOx. The reusability of GOx-mcNF was confirmed by maintaining 70.5 % of its initial activity until the eighth cycle. A limit of detection as 0.7 µM with a wide linear range (0–1000 µM) and the recovery rate of human serum glucose as 97.02–105.9 % were attained. Furthermore, GOx-mcNFs displayed an antibacterial effect due to the generation of ROS by peroxidase-like nanozymes. In vitro antibacterial assays displayed inactivation rates of 99.6 % and 98.8 % against S.aureus and E.coli with 75 μg mL−1 and 750 μg mL−1 of GOx-mcNFs, respectively. Consequently, bi-functional GOx-mcNF provides a promising strategy for glucose biosensing and antibacterial applications.
{"title":"Synthesis and characterization of glucose oxidase bimetallic hybrid magnetic nanoflowers for a sensitive glucose biosensor and an effective antibacterial agent","authors":"Samira Iranmanesh, Arastoo Badoei-Dalfard, Zahra Karami","doi":"10.1016/j.bej.2024.109583","DOIUrl":"10.1016/j.bej.2024.109583","url":null,"abstract":"<div><div>Herein, glucose oxidase was immobilized into bimetallic (Cu<sup>2+</sup>, Zn<sup>2+</sup>) hybrid magnetic nanoflowers (GOx-mcNFs) to perform as a more stable glucose biosensor and antibacterial material. Morphological characterizations of GOx-mcNFs verified the construction of nanoflowers through metal ions-GOx coordination. The encapsulation yield was obtained as 93.5 %, where the activity recovery was 650.3 %. Magnetic nanoparticles also enhanced the peroxidase-like activity of metal-phosphates and caused GOx-mcNFs to show excellent peroxidase-like properties with the apparent <em>K</em><sub><em>m</em></sub> values of 0.19 mM and 0.267 mM for H2O2 and TMB, respectively. Thermodynamic studies revealed a 16.5 kJ mol<sup>−1</sup> higher activation energy of denaturation for GOx-mcNFs over the free GOx. The reusability of GOx-mcNF was confirmed by maintaining 70.5 % of its initial activity until the eighth cycle. A limit of detection as 0.7 µM with a wide linear range (0–1000 µM) and the recovery rate of human serum glucose as 97.02–105.9 % were attained. Furthermore, GOx-mcNFs displayed an antibacterial effect due to the generation of ROS by peroxidase-like nanozymes. <em>In vitro</em> antibacterial assays displayed inactivation rates of 99.6 % and 98.8 % against <em>S.aureus</em> and <em>E.coli</em> with 75 μg mL<sup>−1</sup> and 750 μg mL<sup>−1</sup> of GOx-mcNFs, respectively. Consequently, bi-functional GOx-mcNF provides a promising strategy for glucose biosensing and antibacterial applications.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"214 ","pages":"Article 109583"},"PeriodicalIF":3.7,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142704601","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-19DOI: 10.1016/j.bej.2024.109584
Wenbo Li , Xin Yan , Wenli Xia , Linguo Zhao , Jianjun Pei
Prenylated flavonoids are the primary modification of flavonoids and exhibit a diverse range of physiological activities. In this study, a co-immobilization of two-enzyme cascade was developed to regenerate dimethylallyl diphosphate (DMAPP). Shigella flexneri promiscuous kinase (SfPK) and Methanolobus tindarius isopentenyl phosphate kinase (MtIPK) were immobilized onto carboxymethyl cellulose magnetic nanoparticles (CMN) with a maximum load of 0.35 mg/mg and 0.28 mg/mg, respectively. The optimal activity of CMN-SfPK and CMN-MtIPK were at pH 9.5 and 55°C, and pH 7.0 and 35 °C, respectively. CMN-SfPK and CMN-MtIPK exhibited superior catalytic efficiency compared to free enzymes. CMN-SfPK was coupled with CMN-MtIPK to develop an efficient DMAPP regeneration system from prenol. Subsequently, SfPK, MtIPK and Aspergillus fumigatus prenyltransferase (AfPT) were co-immobilized on CMN to form CMN-SfPK-MtIPK-AfPT (CSMA) according to the optimal ratio. The 3’-C-prenylnaringenin production rate in CSMA reached 0.37 mmol/L/h, which was 1.85 times that of single-immobilized enzymes. Finally, the total production and production rate of 3’-C-prenylnaringenin in CSMA reached 2.55 mM and 0.255 mmol/L/h with 10 cycles. Therefore, the method described herein for efficient production of DMAPP and 3’-C-prenylnaringenin by using co-immobilized enzymes can be widely used for the prenylation of flavonoids.
{"title":"Development of an efficient dimethylallyl diphosphate regeneration system by a co-immobilization of multi-enzyme cascade for the one-pot synthesis of prenylated flavonoids","authors":"Wenbo Li , Xin Yan , Wenli Xia , Linguo Zhao , Jianjun Pei","doi":"10.1016/j.bej.2024.109584","DOIUrl":"10.1016/j.bej.2024.109584","url":null,"abstract":"<div><div>Prenylated flavonoids are the primary modification of flavonoids and exhibit a diverse range of physiological activities. In this study, a co-immobilization of two-enzyme cascade was developed to regenerate dimethylallyl diphosphate (DMAPP). <em>Shigella flexneri</em> promiscuous kinase (SfPK) and <em>Methanolobus tindarius</em> isopentenyl phosphate kinase (MtIPK) were immobilized onto carboxymethyl cellulose magnetic nanoparticles (CMN) with a maximum load of 0.35 mg/mg and 0.28 mg/mg, respectively. The optimal activity of CMN-SfPK and CMN-MtIPK were at pH 9.5 and 55°C, and pH 7.0 and 35 °C, respectively. CMN-SfPK and CMN-MtIPK exhibited superior catalytic efficiency compared to free enzymes. CMN-SfPK was coupled with CMN-MtIPK to develop an efficient DMAPP regeneration system from prenol. Subsequently, SfPK, MtIPK and <em>Aspergillus fumigatus</em> prenyltransferase (AfPT) were co-immobilized on CMN to form CMN-SfPK-MtIPK-AfPT (CSMA) according to the optimal ratio. The 3’-C-prenylnaringenin production rate in CSMA reached 0.37 mmol/L/h, which was 1.85 times that of single-immobilized enzymes. Finally, the total production and production rate of 3’-C-prenylnaringenin in CSMA reached 2.55 mM and 0.255 mmol/L/h with 10 cycles. Therefore, the method described herein for efficient production of DMAPP and 3’-C-prenylnaringenin by using co-immobilized enzymes can be widely used for the prenylation of flavonoids.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"214 ","pages":"Article 109584"},"PeriodicalIF":3.7,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142704600","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
This study investigated the impact of fixed combinations of three flocculants and four buoy-beads under two cultivation systems on the harvesting efficiency, as well as the environmental performance in different scenarios. Results showed that the harvesting efficiency exhibited a tendency of initially increasing and then decreasing with rising concentrations of buoy-beads and flocculants, with an optimal harvesting efficiency of 98.03 %. Life cycle assessment (LCA) compared the environmental performance of five scenarios. Ecotoxicity Soil Chronic (ESC) and Aquatic Eutrophication EP(P) (AEP(P)) were major environmental impacts. The scenario employing re-frying oil emulsion (RFOE) and aluminum sulfate flocculation (R+A) contributed significantly to Human Toxicity Water (HTW) and Aquatic Eutrophication EP(N) (AEP(N)) with normalized values of 0.0137 and 0.0147, respectively. In the assessment of Global Warming Potential (GWP), R+A was responsible for a high amount of Greenhouse Gas (GHG) emissions (2.826 kg CO2 eq/100 g of dry algal biomass in photobioreactor (PBR) and 2.917 kg CO2 eq/ 100 g of dry algal biomass in open raceway ponds (ORP)). Notably, sodium alginate microspheres (SAMs) and aluminum sulfate flocculation (S+A) was considered a more environmentally favorable option, 0.773 kg CO2 eq and 0.864 kg CO2 eq GHG emissions of PBR and ORP, respectively. Furthermore, the less GHG emissions of PBR than ORP, making it a more effective solution for reducing emissions and mitigating global warming trends.
{"title":"Assessing the environmental footprint of microalgae biofuel production: A comparative analysis of cultivation and harvesting scenarios","authors":"Yue Wang, Hao Wen, Meili Wu, Xu Liu, Hongwei Yin, Wei Qin, Xichen Zheng, Jia He, Kemin Wei, Xiaomin Kong, Shuhui Liang","doi":"10.1016/j.bej.2024.109571","DOIUrl":"10.1016/j.bej.2024.109571","url":null,"abstract":"<div><div>This study investigated the impact of fixed combinations of three flocculants and four buoy-beads under two cultivation systems on the harvesting efficiency, as well as the environmental performance in different scenarios. Results showed that the harvesting efficiency exhibited a tendency of initially increasing and then decreasing with rising concentrations of buoy-beads and flocculants, with an optimal harvesting efficiency of 98.03 %. Life cycle assessment (LCA) compared the environmental performance of five scenarios. Ecotoxicity Soil Chronic (ESC) and Aquatic Eutrophication EP(P) (AEP(P)) were major environmental impacts. The scenario employing re-frying oil emulsion (RFOE) and aluminum sulfate flocculation (R+A) contributed significantly to Human Toxicity Water (HTW) and Aquatic Eutrophication EP(N) (AEP(N)) with normalized values of 0.0137 and 0.0147, respectively. In the assessment of Global Warming Potential (GWP), R+A was responsible for a high amount of Greenhouse Gas (GHG) emissions (2.826 kg CO<sub>2</sub> eq/100 g of dry algal biomass in photobioreactor (PBR) and 2.917 kg CO<sub>2</sub> eq/ 100 g of dry algal biomass in open raceway ponds (ORP)). Notably, sodium alginate microspheres (SAMs) and aluminum sulfate flocculation (S+A) was considered a more environmentally favorable option, 0.773 kg CO<sub>2</sub> eq and 0.864 kg CO<sub>2</sub> eq GHG emissions of PBR and ORP, respectively. Furthermore, the less GHG emissions of PBR than ORP, making it a more effective solution for reducing emissions and mitigating global warming trends.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"213 ","pages":"Article 109571"},"PeriodicalIF":3.7,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142655024","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-14DOI: 10.1016/j.bej.2024.109559
Min Gao , Yuan Shen , Yao Peng , Feiyang Tan , Yingshun Lv , Changjie Zhu , Yaxin Guo , Xuan Liu
Coal chemical wastewater (CCW) represents a type of recalcitrant organic wastewater characterized by its intricate composition and high concentration of pollutants, posing a severe threat to global aquatic environments and public health. This study focuses on the degradation of pyridine, a notoriously persistent organic pollutant in CCW, through the identification and application of two highly efficient pyridine-degrading bacterial strains: Alishewanella fetalis (Al-f3) and Exiguobacterium profundum (Ex-p). These strains were immobilized using a polyvinyl alcohol-sodium alginate (PVA-SA) matrix to investigate their bioaugmentation mechanisms in the pyridine degradation process. The findings indicate that strains Al-f3 and Ex-p achieved degradation rates of 95.94 % and 97.83 %, respectively, for an initial pyridine concentration of 200 mg/L at 96 hours. When strains Al-f3 and Ex-p were mixed in equal proportions and immobilized within PVA/SA beads, a degradation rate of 81.06 % was reached within 48 hours, with the efficiency increasing significantly by 96 hours. This enhancement is attributed primarily to the marked increase in enzymatic activity post-immobilization, achieving 17.13 μmol/mg·min, and the elevated secretion of extracellular proteins and polysaccharides, measured at 3.47 mg/L and 1.03 mg/L respectively within 48 hours. Notably, in the immobilized mixed culture system, the total organic carbon (TOC) was reduced to a mere 0.03 mg/L within 72 hours, with a removal rate of 92.31 %. These outcomes not only demonstrate the bioaugmentation role of the immobilized mixed strains in degrading pyridine but also offer novel solutions for the biodegradation of other organic contaminants in CCW, thereby enhancing the treatment efficiency of such wastewater. Given their large specific surface area and cost-effectiveness, PVA/SA immobilization matrices serve as efficient biocarriers in the biodegradation processes for treating CCW. This research provides innovative strategies and methods for the biotreatment of recalcitrant industrial wastewater.
{"title":"Bioenhancement mechanism of PVA-SA immobilized composite strains Alishewanella fetalis and Exiguobacterium profundum in pyridine degradation","authors":"Min Gao , Yuan Shen , Yao Peng , Feiyang Tan , Yingshun Lv , Changjie Zhu , Yaxin Guo , Xuan Liu","doi":"10.1016/j.bej.2024.109559","DOIUrl":"10.1016/j.bej.2024.109559","url":null,"abstract":"<div><div>Coal chemical wastewater (CCW) represents a type of recalcitrant organic wastewater characterized by its intricate composition and high concentration of pollutants, posing a severe threat to global aquatic environments and public health. This study focuses on the degradation of pyridine, a notoriously persistent organic pollutant in CCW, through the identification and application of two highly efficient pyridine-degrading bacterial strains: <em>Alishewanella fetalis</em> (Al-f3) and <em>Exiguobacterium profundum</em> (Ex-p). These strains were immobilized using a polyvinyl alcohol-sodium alginate (PVA-SA) matrix to investigate their bioaugmentation mechanisms in the pyridine degradation process. The findings indicate that strains Al-f3 and Ex-p achieved degradation rates of 95.94 % and 97.83 %, respectively, for an initial pyridine concentration of 200 mg/L at 96 hours. When strains Al-f3 and Ex-p were mixed in equal proportions and immobilized within PVA/SA beads, a degradation rate of 81.06 % was reached within 48 hours, with the efficiency increasing significantly by 96 hours. This enhancement is attributed primarily to the marked increase in enzymatic activity post-immobilization, achieving 17.13 μmol/mg·min, and the elevated secretion of extracellular proteins and polysaccharides, measured at 3.47 mg/L and 1.03 mg/L respectively within 48 hours. Notably, in the immobilized mixed culture system, the total organic carbon (TOC) was reduced to a mere 0.03 mg/L within 72 hours, with a removal rate of 92.31 %. These outcomes not only demonstrate the bioaugmentation role of the immobilized mixed strains in degrading pyridine but also offer novel solutions for the biodegradation of other organic contaminants in CCW, thereby enhancing the treatment efficiency of such wastewater. Given their large specific surface area and cost-effectiveness, PVA/SA immobilization matrices serve as efficient biocarriers in the biodegradation processes for treating CCW. This research provides innovative strategies and methods for the biotreatment of recalcitrant industrial wastewater.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"213 ","pages":"Article 109559"},"PeriodicalIF":3.7,"publicationDate":"2024-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142654798","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-14DOI: 10.1016/j.bej.2024.109573
Huaimin Wang , Amelia R. Bergeson , Hongyuan Lu , Daniel J. Acosta , Ashli J. Silvera , Ryan E. Dittoe , Jessica L.M. Lam , Larissa G.S. Aspiras , August K. Longo , Nathaniel A. Lynd , Hal S. Alper
This study investigates the impact of a specific mutation, designated as the lysine mutation (N233K), on the hydrolytic activity and thermostability of Poly(ethylene terephthalate)-hydrolyzing enzymes (PHEs). This mutation was originally predicted in our prior study using the wild-type I. sakaiensis PETase as a scaffold as a mutation that potentially increases hydrolytic activity of the enzyme. To demonstrate the lysine mutation portability across different PHEs, six PHEs, each containing a mutation equivalent to N233K in PETase, were tested and exhibited improved terephthalic acid (TPA) and mono-(2-hydroxyethyl) terephthalate acid (MHET) monomers release, ranging from 1.05-fold to 5.88-fold relative to the unmutated PHEs. PHL7R205K showed a total of 16.3 mM TPA and MHET monomers release from a plastic disc (at 40 ºC, 72 h) which was 7.1-fold to 18.7-fold higher than all other tested lysine mutants. Finally, kinetic modeling was performed under ambient temperature conditions for FAST-PETase and Z1-PETaseC233K, enabling a benchmark for selecting enzymes for environment remediation and in vivo PET biorecycling.
{"title":"Evaluating the effectiveness of a lysine mutation and its portability across different Poly(ethylene terephthalate)-hydrolyzing enzymes","authors":"Huaimin Wang , Amelia R. Bergeson , Hongyuan Lu , Daniel J. Acosta , Ashli J. Silvera , Ryan E. Dittoe , Jessica L.M. Lam , Larissa G.S. Aspiras , August K. Longo , Nathaniel A. Lynd , Hal S. Alper","doi":"10.1016/j.bej.2024.109573","DOIUrl":"10.1016/j.bej.2024.109573","url":null,"abstract":"<div><div>This study investigates the impact of a specific mutation, designated as the lysine mutation (N233K), on the hydrolytic activity and thermostability of Poly(ethylene terephthalate)-hydrolyzing enzymes (PHEs). This mutation was originally predicted in our prior study using the wild-type <em>I. sakaiensis</em> PETase as a scaffold as a mutation that potentially increases hydrolytic activity of the enzyme. To demonstrate the lysine mutation portability across different PHEs, six PHEs, each containing a mutation equivalent to N233K in PETase, were tested and exhibited improved terephthalic acid (TPA) and mono-(2-hydroxyethyl) terephthalate acid (MHET) monomers release, ranging from 1.05-fold to 5.88-fold relative to the unmutated PHEs. PHL7<sup>R205K</sup> showed a total of 16.3 mM TPA and MHET monomers release from a plastic disc (at 40 ºC, 72 h) which was 7.1-fold to 18.7-fold higher than all other tested lysine mutants. Finally, kinetic modeling was performed under ambient temperature conditions for FAST-PETase and Z1-PETase<sup>C233K</sup>, enabling a benchmark for selecting enzymes for environment remediation and <em>in vivo</em> PET biorecycling.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"214 ","pages":"Article 109573"},"PeriodicalIF":3.7,"publicationDate":"2024-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142704599","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-14DOI: 10.1016/j.bej.2024.109562
Linjiao Ren , Jingtong Sun , Shilin Ma , Diankang Wang , Rubin Qi , Pei Zhang , Qingfang Zhang , Zirui Qin , Liying Jiang
Lead ion residues pose potential health risks to humans. To develop a cost-effective and user-friendly portable lead ion detection method, a novel lateral flow test strip was designed and fabricated based on the G-quadruplex structure. Initially, G4 and its complementary strand antiG4 are in a double-stranded form in the assay solution. After adding lead ions, these ions compete with antiG4 to bind to G4 and form G-quadruplex structures, thus releasing a large number of antiG4 single strands. Through base pairing, one end of the antiG4 was linked to a recognition element containing gold nanoparticles, while the other end was captured by a test line probe, resulting in a red band on the test line. The color change of the test line was positively correlated with the concentration of lead ions. Results showed that by observing the color change of the test line under optimized experimental conditions, lead ion concentration could be detected with a visual detection limit of 20 nM. Quantitative analysis using ImageJ software indicated that the test strip had a linear detection range of 10–2000 nM, with a detection limit of 7.32 nM and significant specificity. The recovery rate in bottled drinking water ranged from 91.19 % to 126.04 %, providing a portable and simple new method for on-site detection of residual lead ions in water environments.
{"title":"A novel lateral flow assay for lead ion detection based on G-quadruplex","authors":"Linjiao Ren , Jingtong Sun , Shilin Ma , Diankang Wang , Rubin Qi , Pei Zhang , Qingfang Zhang , Zirui Qin , Liying Jiang","doi":"10.1016/j.bej.2024.109562","DOIUrl":"10.1016/j.bej.2024.109562","url":null,"abstract":"<div><div>Lead ion residues pose potential health risks to humans. To develop a cost-effective and user-friendly portable lead ion detection method, a novel lateral flow test strip was designed and fabricated based on the G-quadruplex structure. Initially, G4 and its complementary strand antiG4 are in a double-stranded form in the assay solution. After adding lead ions, these ions compete with antiG4 to bind to G4 and form G-quadruplex structures, thus releasing a large number of antiG4 single strands. Through base pairing, one end of the antiG4 was linked to a recognition element containing gold nanoparticles, while the other end was captured by a test line probe, resulting in a red band on the test line. The color change of the test line was positively correlated with the concentration of lead ions. Results showed that by observing the color change of the test line under optimized experimental conditions, lead ion concentration could be detected with a visual detection limit of 20 nM. Quantitative analysis using ImageJ software indicated that the test strip had a linear detection range of 10–2000 nM, with a detection limit of 7.32 nM and significant specificity. The recovery rate in bottled drinking water ranged from 91.19 % to 126.04 %, providing a portable and simple new method for on-site detection of residual lead ions in water environments.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"213 ","pages":"Article 109562"},"PeriodicalIF":3.7,"publicationDate":"2024-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142654797","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-14DOI: 10.1016/j.bej.2024.109566
Xiaoni Zhu, Yanbin Feng, Mingdong Wang, Shang Li, Hongfei Li, Lin Liu, Song Xue
Ferulic acid decarboxylase (Fdc1) catalyzes the decarboxylation of ferulic acid derivatives from lignin with broad substrate spectra. However, the catalytic efficiency of Fdc1 across non-natural substrates with bulky substituents attached to the benzene ring limits its application. Here, an integrated strategy was developed to engineer ScFdc1 from Saccharomyces cerevisiae, specifically targeting 4-acetoxycinnamic acid for the production of 4-acetoxystyrene, which was widely used in photoresists monomers. The strategy entailed the integrating two structural aspects of the enzyme, the conformation of the access tunnel and the substrate binding pocket. Six mutants from a 1248-variant library with significant impacts on enzyme performance were identified. Integration of the two aspects of the mutants achieved the ScFdc1_F397V/I398L/T438P/P441V variant, exhibiting over an 11.8-fold activity improvement towards 4-acetoxycinnamic acid. The variant adopts an open tunnel conformation against that the closed state in the wild type, as revealed by structural analysis with the minimal distance at the bottleneck of the tunnel increased from 0.7 to 1.4 nm. Additionally, a reshaped binding pocket is identified which facilitates the substrate accessibility and binding affinity confirmed by enzymatic and molecular dynamics analysis. The results not only provide strategies for enzyme engineering through the accumulation of beneficial mutants, but also offer a promising route for sustainable styrene derivative production.
{"title":"Integrated strategies for engineering ferulic acid decarboxylase with tunnel conformation and substrate pocket for adapting non-natural substrates","authors":"Xiaoni Zhu, Yanbin Feng, Mingdong Wang, Shang Li, Hongfei Li, Lin Liu, Song Xue","doi":"10.1016/j.bej.2024.109566","DOIUrl":"10.1016/j.bej.2024.109566","url":null,"abstract":"<div><div>Ferulic acid decarboxylase (Fdc1) catalyzes the decarboxylation of ferulic acid derivatives from lignin with broad substrate spectra. However, the catalytic efficiency of Fdc1 across non-natural substrates with bulky substituents attached to the benzene ring limits its application. Here, an integrated strategy was developed to engineer ScFdc1 from <em>Saccharomyces cerevisiae</em>, specifically targeting 4-acetoxycinnamic acid for the production of 4-acetoxystyrene, which was widely used in photoresists monomers. The strategy entailed the integrating two structural aspects of the enzyme, the conformation of the access tunnel and the substrate binding pocket. Six mutants from a 1248-variant library with significant impacts on enzyme performance were identified. Integration of the two aspects of the mutants achieved the ScFdc1_F397V/I398L/T438P/P441V variant, exhibiting over an 11.8-fold activity improvement towards 4-acetoxycinnamic acid. The variant adopts an open tunnel conformation against that the closed state in the wild type, as revealed by structural analysis with the minimal distance at the bottleneck of the tunnel increased from 0.7 to 1.4 nm. Additionally, a reshaped binding pocket is identified which facilitates the substrate accessibility and binding affinity confirmed by enzymatic and molecular dynamics analysis. The results not only provide strategies for enzyme engineering through the accumulation of beneficial mutants, but also offer a promising route for sustainable styrene derivative production.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"213 ","pages":"Article 109566"},"PeriodicalIF":3.7,"publicationDate":"2024-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142654705","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-13DOI: 10.1016/j.bej.2024.109570
Shen-Yuan Yang , Jia-Yih Lin , Pei-Rong Li , Nguyen The Duc Hanh , Penjit Srinophakun , Bing-Lan Liu , Chen-Yaw Chiu , I-Son Ng , Kuei-Hsiang Chen , Yu-Kaung Chang
As industrial activity rises, atmospheric carbon dioxide levels have significantly contributed to global warming and climate change. Developing effective carbon dioxide capture technologies is imperative to mitigate these effects. Carbonic anhydrase (CA) enzymes represent one of the most promising solutions due to their rapid reaction rates, environmental safety, and efficiency in facilitating CO2 conversion. This study takes a novel approach by investigating the immobilization of CA on functionalized polyacrylonitrile (PAN) nanofiber membranes to enhance CO2 conversion and mineralization. PAN nanofibers were fabricated via electrospinning and chemically modified to introduce carboxylic groups, resulting in NM-COOH nanofiber membranes. In addition, amine groups from chitosan (CS) were incorporated to form NM-COOH-CS nanofiber membranes. The immobilization of CA on these membranes revealed that covalent attachment through NM-COOH significantly enhances catalytic activity compared to physical attachment methods. The NM-COOH-CA membrane exhibited superior performance, achieving efficient conversion of CO2 into HCO3− and promoting CaCO3 mineralization. It reached a precipitation efficiency of 11.77 mg CaCO3 per gram of membrane-active unit (WAU) and maintained 63.12 % of its initial CaCO3 production over five cycles over five weeks. This study presents a novel, eco-friendly approach to greenhouse gas reduction, emphasizing the effectiveness of CA-immobilized nanofiber membranes. The findings mainly highlight the advantages of carboxylic functional groups in enhancing CA performance, paving the way for future research in carbon capture technologies.
随着工业活动的增加，大气中的二氧化碳含量大大加剧了全球变暖和气候变化。为减轻这些影响，开发有效的二氧化碳捕获技术势在必行。碳酸酐酶（CA）因其快速的反应速度、环境安全性和促进二氧化碳转化的效率而成为最有前途的解决方案之一。本研究采用一种新方法，研究如何将碳酸酐酶固定在功能化聚丙烯腈（PAN）纳米纤维膜上，以提高二氧化碳的转化和矿化。通过电纺丝制造 PAN 纳米纤维，并对其进行化学改性以引入羧基，从而制成 NM-COOH 纳米纤维膜。此外，还加入了壳聚糖（CS）中的胺基团，形成了 NM-COOH-CS 纳米纤维膜。将 CA 固定在这些膜上的结果表明，与物理附着方法相比，通过 NM-COOH 进行共价附着能显著提高催化活性。NM-COOH-CA 膜性能优越，能将 CO2 高效转化为 HCO3-，并促进 CaCO3 矿化。它的沉淀效率达到每克膜活性单元（WAU）11.77 毫克 CaCO3，并在五周的五个循环中保持了其初始 CaCO3 产量的 63.12%。这项研究提出了一种减少温室气体的新型环保方法，强调了 CA 固定化纳米纤维膜的有效性。研究结果主要强调了羧基官能团在提高 CA 性能方面的优势，为未来碳捕集技术的研究铺平了道路。
{"title":"Functionalized polyacrylonitrile nanofiber membranes with carbonic anhydrase for enhanced carbon dioxide capture and conversion: A performance study","authors":"Shen-Yuan Yang , Jia-Yih Lin , Pei-Rong Li , Nguyen The Duc Hanh , Penjit Srinophakun , Bing-Lan Liu , Chen-Yaw Chiu , I-Son Ng , Kuei-Hsiang Chen , Yu-Kaung Chang","doi":"10.1016/j.bej.2024.109570","DOIUrl":"10.1016/j.bej.2024.109570","url":null,"abstract":"<div><div>As industrial activity rises, atmospheric carbon dioxide levels have significantly contributed to global warming and climate change. Developing effective carbon dioxide capture technologies is imperative to mitigate these effects. Carbonic anhydrase (CA) enzymes represent one of the most promising solutions due to their rapid reaction rates, environmental safety, and efficiency in facilitating CO<sub>2</sub> conversion. This study takes a novel approach by investigating the immobilization of CA on functionalized polyacrylonitrile (PAN) nanofiber membranes to enhance CO<sub>2</sub> conversion and mineralization. PAN nanofibers were fabricated via electrospinning and chemically modified to introduce carboxylic groups, resulting in NM-COOH nanofiber membranes. In addition, amine groups from chitosan (CS) were incorporated to form NM-COOH-CS nanofiber membranes. The immobilization of CA on these membranes revealed that covalent attachment through NM-COOH significantly enhances catalytic activity compared to physical attachment methods. The NM-COOH-CA membrane exhibited superior performance, achieving efficient conversion of CO<sub>2</sub> into HCO<sub>3</sub><sup>−</sup> and promoting CaCO<sub>3</sub> mineralization. It reached a precipitation efficiency of 11.77 mg CaCO<sub>3</sub> per gram of membrane-active unit (WAU) and maintained 63.12 % of its initial CaCO<sub>3</sub> production over five cycles over five weeks. This study presents a novel, eco-friendly approach to greenhouse gas reduction, emphasizing the effectiveness of CA-immobilized nanofiber membranes. The findings mainly highlight the advantages of carboxylic functional groups in enhancing CA performance, paving the way for future research in carbon capture technologies.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"213 ","pages":"Article 109570"},"PeriodicalIF":3.7,"publicationDate":"2024-11-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142654706","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-12DOI: 10.1016/j.bej.2024.109572
Yiying Huo , Pan Feng , Haoran Bi , Kai Wang , Yang Zhang , Yunming Fang , Meng Wang , Tianwei Tan
Terpenoids are widely applied in pharmaceuticals, fragrances and biofuels. Acetyl-CoA, as a crucial precursor in the mevalonate pathway, is stringently regulated intracellularly, thereby limiting the biosynthesis of terpenoids. This study develops a synergistic acetyl-CoA augmentation strategy (SATS) to establish a robust platform for terpenoid biosynthesis in Saccharomyces cerevisiae. Optimizing coenzyme A biosynthesis via global regulation of the pantothenic acid module improved intracellular acetyl-CoA levels by 3.26-fold. Enhancing acetyl phosphate supply was achieved by constructing a PPP-PK-PTA pathway, which improved intracellular acetyl-CoA by 1.92-fold. By combining these techniques, acetyl-CoA levels in the SATS-engineered strain increased by 6.03-fold. The titer of amorpha-4,11-diene, a representative terpenoid, rose by 37.77-fold to 188.45 mg/L in shake-flask fermentation and reached 13 g/L in a 5 L bioreactor. Furthermore, the modified strain exhibited enhanced production of other terpenoids. Our research indicates SATS is an effective approach for synthesizing terpenoids and other acetyl-CoA-derived compounds, demonstrating broad applicability.
{"title":"Synergistic acetyl-CoA augmentation strategy (SATS) for improved terpenoid biosynthesis in Saccharomyces cerevisiae","authors":"Yiying Huo , Pan Feng , Haoran Bi , Kai Wang , Yang Zhang , Yunming Fang , Meng Wang , Tianwei Tan","doi":"10.1016/j.bej.2024.109572","DOIUrl":"10.1016/j.bej.2024.109572","url":null,"abstract":"<div><div>Terpenoids are widely applied in pharmaceuticals, fragrances and biofuels. Acetyl-CoA, as a crucial precursor in the mevalonate pathway, is stringently regulated intracellularly, thereby limiting the biosynthesis of terpenoids. This study develops a synergistic acetyl-CoA augmentation strategy (SATS) to establish a robust platform for terpenoid biosynthesis in <em>Saccharomyces cerevisiae</em>. Optimizing coenzyme A biosynthesis via global regulation of the pantothenic acid module improved intracellular acetyl-CoA levels by 3.26-fold. Enhancing acetyl phosphate supply was achieved by constructing a PPP-PK-PTA pathway, which improved intracellular acetyl-CoA by 1.92-fold. By combining these techniques, acetyl-CoA levels in the SATS-engineered strain increased by 6.03-fold. The titer of amorpha-4,11-diene, a representative terpenoid, rose by 37.77-fold to 188.45 mg/L in shake-flask fermentation and reached 13 g/L in a 5 L bioreactor. Furthermore, the modified strain exhibited enhanced production of other terpenoids. Our research indicates SATS is an effective approach for synthesizing terpenoids and other acetyl-CoA-derived compounds, demonstrating broad applicability.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"213 ","pages":"Article 109572"},"PeriodicalIF":3.7,"publicationDate":"2024-11-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142655248","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-12DOI: 10.1016/j.bej.2024.109564
Huaiyu Duan , Xingqing Zhao , Chao Xu , Du Zhang , Wei Gu , Rucheng Wang , Xiancai Lu
The aim of this study was to explore the improvement of lithium leaching from lepidolite by microbial co-culture, focusing on the synergistic effect of different structural fungi in improving the leaching performance of biological systems. The results showed that in the single leaching experiment, the multicellular fungi and the unicellular yeast showed weak effects. Multicellular fungus is limited by insufficient EPS secretion, while unicellular yeast is non-mycelial organisms with weak acid production capacity and limited effect on minerals. However, in the combined leaching experiment, the interspecific collaboration promoted the synthetic and metabolic activity of the two strains, resulting in changes in the type and content of organic acids, polysaccharides, proteins and humus. The content of citric acid reached the highest value of 16.98 g·L−1 at about 22 d, and the EPS secreted by unicellular yeast promoted the mycelium adhesion and mineral wrapping of multicellular fungi. The combined action of the two enhanced the effects of acidification, complexation and mycelium destruction, and improved the leaching effect. This study revealed the synergistic metabolic mechanism of lepidolite leaching by fungi with different structures, verified the effectiveness of microbial co-culture to improve the leaching rate, and provided a basis for industrial application. In addition, the use of co-culture technology has a positive impact on commercial production and environmental protection.
本研究旨在探索微生物共培养对锂从鳞片石中浸出的改善作用，重点关注不同结构真菌在改善生物系统浸出性能方面的协同效应。结果表明，在单一浸出实验中，多细胞真菌和单细胞酵母菌的作用较弱。多细胞真菌受限于 EPS 分泌不足，而单细胞酵母菌是非菌丝生物，产酸能力弱，对矿物质的作用有限。但在联合浸出实验中，种间协作促进了两种菌株的合成代谢活动，使有机酸、多糖、蛋白质和腐殖质的种类和含量发生了变化。柠檬酸的含量在 22 d 左右达到最高值 16.98 g-L-1，单细胞酵母菌分泌的 EPS 促进了多细胞真菌的菌丝粘附和矿物包裹。两者的联合作用增强了酸化、络合和菌丝破坏的效果，提高了浸出效果。该研究揭示了不同结构真菌对鳞片石浸出的协同代谢机制，验证了微生物共培养对提高浸出率的有效性，为工业应用提供了依据。此外，共培养技术的使用还对商业生产和环境保护产生了积极影响。
{"title":"The synergistic interaction of fungi with different structural characteristics improves the leaching of lithium from lepidolite","authors":"Huaiyu Duan , Xingqing Zhao , Chao Xu , Du Zhang , Wei Gu , Rucheng Wang , Xiancai Lu","doi":"10.1016/j.bej.2024.109564","DOIUrl":"10.1016/j.bej.2024.109564","url":null,"abstract":"<div><div>The aim of this study was to explore the improvement of lithium leaching from lepidolite by microbial co-culture, focusing on the synergistic effect of different structural fungi in improving the leaching performance of biological systems. The results showed that in the single leaching experiment, the multicellular fungi and the unicellular yeast showed weak effects. Multicellular fungus is limited by insufficient EPS secretion, while unicellular yeast is non-mycelial organisms with weak acid production capacity and limited effect on minerals. However, in the combined leaching experiment, the interspecific collaboration promoted the synthetic and metabolic activity of the two strains, resulting in changes in the type and content of organic acids, polysaccharides, proteins and humus. The content of citric acid reached the highest value of 16.98 g·L<sup>−1</sup> at about 22 d, and the EPS secreted by unicellular yeast promoted the mycelium adhesion and mineral wrapping of multicellular fungi. The combined action of the two enhanced the effects of acidification, complexation and mycelium destruction, and improved the leaching effect. This study revealed the synergistic metabolic mechanism of lepidolite leaching by fungi with different structures, verified the effectiveness of microbial co-culture to improve the leaching rate, and provided a basis for industrial application. In addition, the use of co-culture technology has a positive impact on commercial production and environmental protection.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"213 ","pages":"Article 109564"},"PeriodicalIF":3.7,"publicationDate":"2024-11-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142655023","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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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