Biochemical Engineering Journal最新文献_第10页

Effective expression of taste-modifying protein miraculin fusing FMDV 2A gene in Chlamydomonas reinhardtii 融合FMDV 2A基因的味觉修饰蛋白miraculin在莱茵衣藻中的有效表达

IF 3.7 3区生物学

Biochemical Engineering Journal Pub Date : 2025-03-06 DOI: 10.1016/j.bej.2025.109706

Chuan Wang, Xiaomei Wang, Ge Rang Lamu, Wei Liang, Yaomei Tian

{"title":"Effective expression of taste-modifying protein miraculin fusing FMDV 2A gene in Chlamydomonas reinhardtii","authors":"Chuan Wang, Xiaomei Wang, Ge Rang Lamu, Wei Liang, Yaomei Tian","doi":"10.1016/j.bej.2025.109706","DOIUrl":"10.1016/j.bej.2025.109706","url":null,"abstract":"<div><div>Miraculin is a taste modification protein derived from plants, which can change sour taste into sweet taste, and has great application potential in food industry. Currently, the extraction of miraculin presented some problems such as cumbersome process and low purity. <em>Chlamydomonas reinhardtii</em> is a potential expression system of exogenous proteins. However, the exogenous genes are integrated randomly in genome of <em>C. reinhardtii</em> during transformation, which leads to significant expression variability among transformants. The 2A peptide from FMDV virus has been proven to effectively improve the expression efficiency of exogenous proteins in multi-gene expression systems through self-cleavage mechanism. In presented study, the expression vector pSP108–2A-miraculin containing 2A-miraculin fusion gene was constructed and transformed into <em>C. reinhardtii</em> by glass bead method. Two positive strains were obtained through bleomycin resistance selecting. The detection results showed that miraculin gene was integrated into the genome of <em>C. reinhardtii</em> cells at the molecular level, and was successfully expressed at the protein level. This study was the first time to express the miraculin in <em>C. reinhardtii</em>, which provided an optional way to obtain miraculin.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"219 ","pages":"Article 109706"},"PeriodicalIF":3.7,"publicationDate":"2025-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143593353","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}

引用次数: 0

Optimization modeling and economics assessment on simultaneous struvite and bioelectricity production from waste nutrient solution in the microbial fuel cell 微生物燃料电池废营养液同时产鸟粪石和生物电的优化建模及经济性评价

IF 3.7 3区生物学

Biochemical Engineering Journal Pub Date : 2025-03-04 DOI: 10.1016/j.bej.2025.109705

Sreelakshmi C , Kiruthika S , Jeyalakshmi R

{"title":"Optimization modeling and economics assessment on simultaneous struvite and bioelectricity production from waste nutrient solution in the microbial fuel cell","authors":"Sreelakshmi C , Kiruthika S , Jeyalakshmi R","doi":"10.1016/j.bej.2025.109705","DOIUrl":"10.1016/j.bej.2025.109705","url":null,"abstract":"<div><div>Several Sustainable Development Goals (SDGs) emphasize community well-being and environmental health. Addressing food and energy demands while ensuring a clean environment remains a global challenge. This study investigates nutrient recovery as struvite and bioelectricity generation from nutrient-rich wastewater using a dual-chamber microbial fuel cell (MFC). Response Surface Methodology with Central Composite Design (CCD) optimized Phosphorus (P) recovery at minimal cost. Simulation parameters, including the N:P ratio, chemical oxygen demand (COD), and cathodic aeration rate, were tested. Results showed that P recovery efficiency was influenced by the N:P ratio and COD, while power density was primarily impacted by COD. Optimized conditions of N:P = 1, COD = 3000 mg/L, aeration = 150 mL/min achieved 90 % P recovery as magnesium ammonium phosphate hexahydrate (struvite, MgNH₄PO₄·6H₂O) with a power density of 860 mW/m². The estimated cost based on CCD simulations was 2.5–2.6 USD/m³ within a 5 % deviation under optimal conditions of N:P = 2, COD = 3000 mg/L, aeration = 150 mL/min. These findings highlight the potential of integrating MFCs into wastewater treatment plants (WTPs) for nutrient recovery, COD reduction, and bioelectricity production. This approach offers a viable solution for sustainable wastewater management but requires careful assessment of Technology Readiness Level (TRL) and System Readiness Level (SRL) for large-scale implementation.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"218 ","pages":"Article 109705"},"PeriodicalIF":3.7,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143550449","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}

引用次数: 0

Biochemical production from sustainable carbon sources by Komagataella phaffii 利用可持续碳源进行生化生产的Komagataella phaffii

IF 3.7 3区生物学

Biochemical Engineering Journal Pub Date : 2025-03-03 DOI: 10.1016/j.bej.2025.109702

Wajeeha A. Raja , Pınar Çalık

{"title":"Biochemical production from sustainable carbon sources by Komagataella phaffii","authors":"Wajeeha A. Raja , Pınar Çalık","doi":"10.1016/j.bej.2025.109702","DOIUrl":"10.1016/j.bej.2025.109702","url":null,"abstract":"<div><div>The use of sustainable carbon sources (SCSs) for biochemical production is needed to facilitate the chemical process industries transition towards the net-zero age and greener economy. The use of SCSs such as crude glycerol, ethanol, methanol, acetate, lactate, and formate offers potential cost reduction and sustainability benefits. However, the integration of SCSs into bioindustries is generally challenged by their low uptake rates, toxicity, and the metabolic stress they exert. Therefore, their transport rates from fermentation medium into the cell and then sequential intracellular reaction rates towards target products need to be fine-tuned. This review begins with the importance of use of SCSs in biochemical process industries and continues with the current state of metabolic engineering strategies employed to optimize biochemical and recombinant protein production. In turn, metabolic engineering strategies to increase the uptake rates of two SCSs, ethanol and acetate, and the carbon fluxes fueling the TCA cycle in <em>Komagataella phaffii</em> (<em>Pichia pastoris</em>) are discussed. Moreover, the latest findings on bioreactor operation condition optimization are compiled with a special emphasis on recombinant protein production. We conclude that the future of <em>K. phaffii</em>-based bioprocessing lies in integrating metabolic and bioreactor engineering with systems biology, while advances in synthetic biology, such as modular genome editing and machine learning for promoter and pathway design, will further refine the metabolic versatility of <em>K. phaffii</em>.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"219 ","pages":"Article 109702"},"PeriodicalIF":3.7,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143592752","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}

引用次数: 0

Rational design based on translation pausing theory significantly enhances the soluble expression and activity of multidomain anti-CD20 fab antibody sequences 基于翻译暂停理论的合理设计显著提高了多结构域抗cd20 fab抗体序列的可溶性表达和活性

IF 3.7 3区生物学

Biochemical Engineering Journal Pub Date : 2025-03-03 DOI: 10.1016/j.bej.2025.109704

Aiping Kong , Shiwen Chen , Wenxi Huang , Xinshan Xie , Qiuling Xie , Sheng Xiong

{"title":"Rational design based on translation pausing theory significantly enhances the soluble expression and activity of multidomain anti-CD20 fab antibody sequences","authors":"Aiping Kong , Shiwen Chen , Wenxi Huang , Xinshan Xie , Qiuling Xie , Sheng Xiong","doi":"10.1016/j.bej.2025.109704","DOIUrl":"10.1016/j.bej.2025.109704","url":null,"abstract":"<div><div>The expression of IgG antibodies in Chinese hamster ovary (CHO) cells has been widely used, but their expression in <em>Escherichia coli</em> remains challenging. In antibody engineering and drug research, the expression and functional study of antibody fragments in <em>E. coli</em> is crucial for early development. However, these fragments often aggregate to form inclusion bodies or exhibit low solubility when expressed in the periplasm, making them difficult to obtain. In this study, we propose a strategy based on the theory of translational pausing, where mutations are introduced into the DNA sequence to alter the translation pausing sites without changing the amino acid sequence, thus promoting correct protein folding and improving the solubility of heterologous proteins in <em>E. coli</em>. We previously successfully optimized the antiviral protein cyanovirin-N (CVN) from cyanobacteria and epoxide hydrolase (EH-Ar) from Agrobacterium, and we attempted to express an anti-CD20 Fab antibody with a quaternary structure to verify whether the translational pausing technique can increase the solubility and expression of multidomain proteins. Compared with the wild-type sequence, the designed coding sequence exhibited greater soluble expression in <em>E. coli</em>, and the expressed Fab antibody retained its activity. This study further demonstrates the importance of translational pausing in the expression of heterologous proteins in bacterial hosts and provides a novel approach for soluble expression and rapid preparation of recombinant small-molecule antibodies.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"218 ","pages":"Article 109704"},"PeriodicalIF":3.7,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143550450","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}

引用次数: 0

Corrigendum to “Scaling iPSC production in different 3D platforms in suspension culture for their use in allogeneic regenerative therapies” [Biochem. Eng. J. 217 (2025) 109672] “在悬浮培养的不同3D平台上扩大iPSC生产，用于异体再生疗法”的更正[Biochem]。Eng。J. 217 (2025) 109672]

IF 3.7 3区生物学

Biochemical Engineering Journal Pub Date : 2025-03-01 DOI: 10.1016/j.bej.2025.109701

Danton Freire-Flores , Nyna Kawles , Pablo Caviedes , Barbara A. Andrews , Juan A. Asenjo

引用次数: 0

Maximizing harvesting of Chlorella vulgaris via calcium oxide nanoparticle-modified fungal pellet using response surface methodology 利用响应面法通过氧化钙纳米颗粒修饰真菌颗粒最大化地收获普通小球藻

IF 3.7 3区生物学

Biochemical Engineering Journal Pub Date : 2025-03-01 DOI: 10.1016/j.bej.2025.109700

Muhammad Hizbullahi Usman , Mohd Farizal Kamaroddin , Mohd Helmi Sani , Aliyu Ibrahim Dabai , Abdulrahman Sani Aliero , Ali El-Rayyes

{"title":"Maximizing harvesting of Chlorella vulgaris via calcium oxide nanoparticle-modified fungal pellet using response surface methodology","authors":"Muhammad Hizbullahi Usman , Mohd Farizal Kamaroddin , Mohd Helmi Sani , Aliyu Ibrahim Dabai , Abdulrahman Sani Aliero , Ali El-Rayyes","doi":"10.1016/j.bej.2025.109700","DOIUrl":"10.1016/j.bej.2025.109700","url":null,"abstract":"<div><div>This study aims to optimize the harvesting of <em>Chlorella vulgaris</em> using calcium oxide nanoparticle modified-<em>Aspergillus pseudonomiae</em> pellets (AP@CaONP). To maximize harvesting efficiency, response surface methodology (RSM-CCD) was applied to statistically optimize key parameters, including mycelial dosage (%), agitation speed (rpm), and pH. Lipid yield from harvested <em>C. vulgaris</em> biomass under optimized conditions was analyzed, and resulting fatty acid methyl esters were identified using Gas Chromatography-Mass Spectrometry (GC-MS). Additionally, the recycling potential of spent medium for recultivation of <em>C. vulgaris</em> was evaluated. Scanning Field Emission Scanning Electron Microscopy (FESEM), Fourier Transform Infrared spectroscopy (FTIR), and zeta potential analyses were conducted to confirm the interaction between AP@CaONP and <em>C. vulgaris</em>. The results show that under optimized conditions (6 % mycelial dosage, 170 rpm, pH 5.5), harvesting efficiency reached 99.24 % in 60 minutes with only a 0.2 % deviation between predicted and experimental values. Lipid yield from harvested biomass reached 26.98 %, and GC-MS revealed fatty acids including C16:0, C16:1, C16:2, C18:0, C18:1, and C18:2. <em>C. vulgaris</em> showed similar growth patterns in fresh and recycled BG-11 media, indicating minimal impact on biomass productivity and reduced water consumption. This optimized AP@CaONP-based harvesting method offers a practical, eco-friendly approach for efficient microalgae biomass recovery, supporting sustainable biodiesel production from <em>Chlorella</em> species.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"219 ","pages":"Article 109700"},"PeriodicalIF":3.7,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143600534","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}

引用次数: 0

Simultaneous enzymatic hydrolysis and bioconversion of deacetylated and disk refined sugarcane bagasse to single-cell protein: An experimental investigation and techno-economic analysis 脱乙酰和圆盘精制甘蔗渣同时酶解和生物转化为单细胞蛋白：实验研究和技术经济分析

IF 3.7 3区生物学

Biochemical Engineering Journal Pub Date : 2025-02-24 DOI: 10.1016/j.bej.2025.109691

Veronica Lourens, Catharine E. Bosman , Abdul M. Petersen , Gerhardt Coetzee, Johann F. Görgens , Eugéne van Rensburg

{"title":"Simultaneous enzymatic hydrolysis and bioconversion of deacetylated and disk refined sugarcane bagasse to single-cell protein: An experimental investigation and techno-economic analysis","authors":"Veronica Lourens, Catharine E. Bosman , Abdul M. Petersen , Gerhardt Coetzee, Johann F. Görgens , Eugéne van Rensburg","doi":"10.1016/j.bej.2025.109691","DOIUrl":"10.1016/j.bej.2025.109691","url":null,"abstract":"<div><div>Rising animal feed costs are driving up animal agriculture and food prices. While single-cell protein (SCP) production from lignocellulosic waste offers a solution, pretreatment of these feedstocks is required. In this study, the SCP yield from sugarcane bagasse pretreated via deacetylation in combination with disk refining (DDR), prior to simultaneous enzymatic hydrolysis and fermentation using <em>P. ostreatus</em> and <em>F. venenatum</em> compared to <em>S. cerevisiae</em>, was investigated. A biorefinery was simulated to investigate the economic feasibility of animal feed enhanced with SCP, using a discounted cashflow analysis. Deacetylation and DDR were effective pretreatment approaches, improving enzymatic digestibility from 9.10 g sugar/100 g dry matter (DM) to 75.75 and 80.14 g sugar/100 g DM, respectively, which was comparable to steam-explosion pretreatment. Pretreated bagasse was successfully converted to SCP through simultaneous enzymatic hydrolysis and bioconversion. However, DDR was preferred for bioconversions using <em>P. ostreatus</em> and <em>F. venenatum</em>, achieving SCP yields of 4.24 ± 0.17 and 3.47 ± 0.33 g protein/100 g DM, respectively. In the ranges investigated, solids loading had a more pronounced effect on the SCP yield than enzyme dosage, irrespective of the microorganism used. However, the minimum selling price of animal feed enhanced with SCP (0.69 US$/kg) remains higher than the market price of animal feeds like soybean meal (0.40 US$/kg). The technology shows promise but can be further optimised.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"218 ","pages":"Article 109691"},"PeriodicalIF":3.7,"publicationDate":"2025-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143520508","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Synergy between horseradish peroxidase HRP and edible gelatin: A simple way to remove phenols and dyes simultaneously from wastewater 辣根过氧化物酶HRP和食用明胶的协同作用：同时从废水中去除酚类和染料的简单方法

IF 3.7 3区生物学

Biochemical Engineering Journal Pub Date : 2025-02-23 DOI: 10.1016/j.bej.2025.109692

Jianling Feng , Huadeng Li , Ke Zheng , Yanbin Zeng , Guanqun Xie , Xiaoxia Wang

{"title":"Synergy between horseradish peroxidase HRP and edible gelatin: A simple way to remove phenols and dyes simultaneously from wastewater","authors":"Jianling Feng , Huadeng Li , Ke Zheng , Yanbin Zeng , Guanqun Xie , Xiaoxia Wang","doi":"10.1016/j.bej.2025.109692","DOIUrl":"10.1016/j.bej.2025.109692","url":null,"abstract":"<div><div>Wastewater containing phenols or dyes is quite common. Given the potential harmful effects of such wastewater on both the environment and human health, there is a continuing demand for efficient, green, and cost-effective methods to remove these pollutants. In this study, edible gelatin was employed as an additive in conjunction with horseradish peroxidase (HRP) to purify wastewater containing phenols and dyes. Under optimized conditions, the combination of HRP and gelatin effectively purified various types of wastewater containing different phenols, achieving removal rates of up to 90 %. Furthermore, this strategy was successfully applied to the degradation of mixed wastewater containing dyes (methyl orange, methylene blue or/and rhodamine B) in the presence of phenol. High removal rates for both phenol and the dyes were also observed and the TOC removal rate reached as high as 94 %. Finally, a removal mechanism involving the HRP/gelatin system was proposed based on UV-Vis detection, particle size analysis, LC-MS, and TOC results. This green and efficient strategy for the removal of phenols and dyes from wastewater represents a valuable contribution to sustainable environmental remediation.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"218 ","pages":"Article 109692"},"PeriodicalIF":3.7,"publicationDate":"2025-02-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143508784","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}

引用次数: 0

Complete scheme for biosuccinic acid production from candy-factory waste with simultaneous fermentation-electrochemical product recovery, and purification 糖厂废弃物同步发酵生产生物琥珀酸的完整方案-电化学产物回收和纯化

IF 3.7 3区生物学

Biochemical Engineering Journal Pub Date : 2025-02-21 DOI: 10.1016/j.bej.2025.109687

Ioannis Zacharopoulos, Antonio Grimalt-Alemany, Christoforos Tsoumanis, Irini Angelidaki

{"title":"Complete scheme for biosuccinic acid production from candy-factory waste with simultaneous fermentation-electrochemical product recovery, and purification","authors":"Ioannis Zacharopoulos, Antonio Grimalt-Alemany, Christoforos Tsoumanis, Irini Angelidaki","doi":"10.1016/j.bej.2025.109687","DOIUrl":"10.1016/j.bej.2025.109687","url":null,"abstract":"<div><div>Succinic acid is a high-value platform chemical with a wide range of industrial applications, making its sustainable production an area of increasing interest. However, a key challenge preventing biologically produced succinic acid from becoming economically competitive with petrochemical-based production lies in the inefficiencies associated with both upstream and downstream processing. In this study, we present a comprehensive platform for the biological production of succinic acid, using industrial waste from candy production as a substrate. This approach integrates upstream and downstream processes through the use of an in-situ electrochemical succinic acid recovery system, significantly simplifying the overall production workflow. As a result, the downstream process was reduced to just five steps, compared to the more complex nine-step processes described in existing literature. The proposed method demonstrated high separation yields of 86 % and a purity level of 92 %, showcasing its efficiency. Furthermore, the in-situ recovery strategy led to a 15 % increase in succinic acid yield, a 10.2 % improvement in overall productivity, and a 13.5 % increase in substrate consumption when compared to traditional batch fermentation processes. These results highlight the potential of this integrated platform to enhance the economic feasibility of biologically produced succinic acid on an industrial scale.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"218 ","pages":"Article 109687"},"PeriodicalIF":3.7,"publicationDate":"2025-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143480488","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}

引用次数: 0

Synthesis of (R)-acetoin by targeted immobilization of acetolactate synthase and acetolactate decarboxylase via the spy/snoop system spy/snoop系统固定化乙酰乳酸合酶和乙酰乳酸脱羧酶合成(R)-乙酰托因

IF 3.7 3区生物学

Biochemical Engineering Journal Pub Date : 2025-02-19 DOI: 10.1016/j.bej.2025.109671

Kexin Shao , Yadong Wang , Peng Chen , Fenghuan Wang

{"title":"Synthesis of (R)-acetoin by targeted immobilization of acetolactate synthase and acetolactate decarboxylase via the spy/snoop system","authors":"Kexin Shao , Yadong Wang , Peng Chen , Fenghuan Wang","doi":"10.1016/j.bej.2025.109671","DOIUrl":"10.1016/j.bej.2025.109671","url":null,"abstract":"<div><div>Acetoin (3-hydroxy-2-butanone) is an important four-carbon platform compound and an intermediate for several high-value-added pharmaceutical compounds. In this study, Catcher-Tag tagged proteins were used for targeted immobilization of acetolactate synthase (ALS) and acetolactate decarboxylase (ALDC) for catalyzing the synthesis of Acetoin from sodium pyruvate in vitro. The study discussed the enzymatic properties of the immobilized enzyme LX@ALS-SpyTag 和 LX@ALDC-SnoopTag, as well as the optimal catalytic conditions and reusability of the immobilized dual enzyme in vitro catalytic system. The immobilized enzyme exhibited higher relative activity across a broader range of temperature and pH intervals than the free enzyme. The in vitro catalytic synthesis of (R)-acetoin from sodium pyruvate using an immobilized dual enzyme system resulted in 99.95 % of the theoretical conversion of (R)-acetoin at 45°C, pH 7.0, and 0.2 M substrate concentration. In addition, the immobilized dual enzyme system had better operational stability than the free enzyme, retaining 69.20 % of the initial total catalytic activity after 5 cycles. In conclusion, this study demonstrated a promising and convenient Catcher-Tag tagged protein immobilization strategy that enables rapid targeted immobilization of tagged proteins directly from crude enzyme solution, providing a novel approach for the construction of in vitro immobilized multi-enzyme complexes.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"217 ","pages":"Article 109671"},"PeriodicalIF":3.7,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143445410","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}

引用次数: 0