ACS Applied Materials & Interfaces
·
2018
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IF 8.097
Zhenning Liu,Bingdi Wang,Shenghui Jin,Zhida Wang,Lei Wang,Song Liang
In this paper, a novel method for the construction of colloidosomes as a microreactor for dual-enzyme cascade biphasic reaction has been reported. A lipase–glucose oxidase (GOx) enzyme pair is employed in this system. A water-soluble enzyme GOx is compartmentalized inside the colloidosomes. A hydrophobic environment-favored enzyme Candida Antarctica lipase B (CalB) is adsorbed on the outer surfaces of the colloidosomes. The catalysis system is set up by introducing these dual-enzyme-immobilized microcapsules into acetic ether. H2O2 is produced in the aqueous phase by the doped GOx, and then H2O2 diffused out of the microcapsules is utilized by CalB to catalyze the oxidation of ethyl acetate. Finally, the formed peracids oxidized N-heteroaromatic in situ. Furthermore, no obvious yield decline is observed in four reaction cycles. Thus, our work provides a new strategy for the design of high-performance biomimicking reactors for multiple enzyme cascade reactions and further expands the potential application area of colloidosomes.
研究领域:
生物化学
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材料科学
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DOI
PubMed
高影响因子
Molecular Catalysis
·
2023
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IF 4.6
Bin Wu,Xiangyun Wang,Bo Yang,Yunjian Ma,Frank Hollmann,Yonghua Wang
In the present work we propose a bienzymatic cascade for the oxyfunctionalisation of cycloalkanes to cyclic alcohols/cyclic ketones. By combining a H 2 O 2 -dependent peroxygenase with a O 2 -consuming and H 2 O 2 -producing alcohol oxidase an overall aerobic oxidation system was established. A convincing proof-of-concept is presented and some current limitations are outlined.
研究领域:
生物化学
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酶工程
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DOI
高影响因子
Biomolecules
·
2024
·Vol. 14
·期 12
·1553
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IF 4.8
Zifu Ni,Linshang Zhang,Azhen Nie,Huan Wang,Xiaoling Wu
As the only naturally occurring β-amino acid, β-alanine has important application prospects in many fields. Driven by the huge demand, biosynthesis is becoming more and more popular as a potential alternative to the chemical synthesis of β-alanine. Although the direct pathway from L-aspartic acid to β-alanine, catalyzed by L-aspartic acid-α-decarboxylase (PanD), is ideal for β-alanine synthesis, it is hindered by the high cost of the substrate and limited economic viability. In this work, a cell-free dual enzyme cascade system based on methylaspartate lyase (EcMAL) and panD was constructed to safely and efficiently synthesize β-alanine using fumarate as a substrate. Taking the previously engineered EcMAL as the target, CgPanD was finally screened as the best candidate through gene mining, sequence alignment, and enzyme property analysis. Finally, under the optimal conditions of 35 °C, pH 8.0, and EcMAL: CgPanD concentration ratio of 1:5, the yield of β-alanine reached 80% theoretical yield within 120 min. This study provides a potential strategy for the biosynthesis of β-alanine, paving the way for future industrial-scale production.
研究领域:
生物化学
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合成生物学
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PubMed
高影响因子
ACS Omega
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2025
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IF 4.3
Tianfu Wu,Wei Song,Jing Wu,Jian Wen,Wanqing Wei,Liming Liu
Chiral amino alcohols are vital in many biologically active molecules and natural products and are crucial building blocks for the pharmaceutical industry. However, the chemosynthesis of chiral amino alcohols, particularly bichiral amino alcohols, faces challenges related to chemoselectivity and enantioselectivity. Here, a dual-enzyme cascade pathway for the synthesis of bichiral amino alcohols through difunctionalization, exhibiting both high chemo- and stereoselectivity, was proposed for the first time. Among them, NADPH-dependent meso-diaminopyrimidate dehydrogenase from Proteus vulgaris (PvDAPDH) exhibited (R)-selective reductive amination activity, converting ketones into chiral amines; natural glucose dehydrogenase from Bacillus megaterium (BmGDH) not only facilitated NADPH regeneration but also catalyzed the reductive hydroxylation of ketones to chiral alcohols. Second, the rational design of the rate-limiting enzyme PvDAPDH generated the mutant M4 with increased specific activity and catalytic efficiency (kcat/Km) for 16a up to 3.2- and 10.3-fold, respectively, compared to the wild type. Additionally, the yield of mutant M4 for 1a–16a increased by 1.1- to 37-fold compared to the wild type. Finally, two high-value bichiral amino alcohols (14b and 16b) were asymmetrically synthesized from the corresponding bietone via a one-pot dual-enzyme cascade, exhibiting excellent stereoselectivity (dr ≥ 98:2). These findings provide a potential biosynthetic pathway for the green synthesis of bichiral amino alcohols and complement existing synthetic methods.
研究领域:
生物化学
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合成生物学
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制药工程
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
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2025
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IF 10.5
Xiao Liang,Zhuobing Zhang,Yilei Du,Ziliang Li,Nana Xu,Jian Gao,Huimin Qin,Liping Du,Song You,Jinshan Li,Weidong Liu,Yanhe Ma
Ochratoxins, toxic mycotoxins present in food and feed, pose a threat to human health due to their various toxic effects. Developing a safe, convenient, and rapid on-site detection tool for monitoring ochratoxins is essential for food and environmental safety, especially in developing countries. Although numerous effective ochratoxin sensing and detection systems are available, most do not combine detoxification with detection capabilities. In this work, an ochratoxin-detoxifying enzyme was cascaded with phenylalanine dehydrogenase to construct a colorimetric biosensor capable of simultaneously detecting and removing Ochratoxin A (OTA) and Ochratoxin B (OTB) contamination. Once the ochratoxins are detoxified into non-toxic OTα/OTβ and phenylalanine, 1-methoxy-5-methylphenazinium methyl sulfate (MPMS) functions as a signal reporter, converting the ochratoxin concentration into a colorimetric signal that gradually shifts from red to green. This allows for real-time visual observation with the naked eye. Additionally, OTA/OTB within a concentration range of 0–50 μg/mL can be quantitatively analyzed using a microplate reader or a smartphone. Under the optimized conditions, the colorimetric biosensor effectively eliminated and detected OTA/OTB at a spiked concentration of 5 μg/L in Chinese liquor samples within 1 h. Hence, the dual-enzyme cascade colorimetric biosensor has been demonstrated to be a safe, portable, rapid, and accurate tool for the detection and elimination of OTA/OTB in food.
研究领域:
生物传感技术
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食品安全
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环境安全
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酶催化
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分析化学
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2024
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IF 7.7
Jingjie Gao,Shaowei Bai,Fan Wang,Liangzhen Yang,Yufeng Hu,Yuhuan Yang,Bing Bai,Zuoming Zhang
Enzymatic glycosylation is an efficient and biocompatible approach to enhance natural product bioavailability. Cellobiose phosphorylase, a novel glycosyltransferase, utilizes 1-phospho-glucose (1-p-Glc) as a glycosyl donor for regioselective glycosylation of various natural substrates. However, the high cost of 1-p-Glc limits the economic feasibility of the process. Thus, a dual-enzyme cascade system involving cellulase Ac Cel9A and cellobiose phosphorylase Cb CBP using a co-immobilization strategy was developed to overcome this challenge. The system utilizes low-cost carboxymethyl cellulose (CMC) for continuous 1-p-Glc production, which was then used in the fluorodeoxy glucose (FDG) glycosylation to generate fluorodeoxy cellobiose (FDC), which potentially traces fungal infections. The compartmentalized co-immobilization of the two enzymes within the internal and external regions of a porous zeolitic imidazolate framework-8 (ZIF-8) carrier enhanced the overall stability of the dual-enzyme system. The immobilized enzymes retained approximately 63.3 % activity after seven reuse cycles and 74 % catalytic efficiency after 12 days of storage at room temperature. Therefore, the developed co-immobilized multi-enzyme system holds significant potential for industrial biocatalysis applications.
研究领域:
生物化学
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工业酶学
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合成生物学
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DOI
PubMed
高影响因子
Bioactive Materials
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2025
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IF 20.3
Qi Zhang,Zhe Zheng,Yiting Zhao,Qing Wu,Chu Wu,Xiuli Wang,Xia Wang,Qigang Wang,Peiru Wang
The transformation of O 2 is primarily facilitated by the catalytic action of redox enzymes, which play a pivotal role in sustaining cellular energy metabolism and redox balance. Bioinspired by O 2 distribution and ROS regulation related to cascade biocatalytic process, a D-peptide NapG D F D F D Y engineered hydrogel has been constructed with encapsulated dual-enzyme superoxide (SOD) and chloroperoxidase (CPO) cascade catalytic circuit and co-assembled photosensitizer of 5-aminolevulinic acid molecules (ALA). Multimodal oxygen modulation has been conducted by the concurrent oxygen generation via SOD-catalyzed ⋅O 2 − dismutation and oxygen consumption for 1 O 2 production by CPO and ALA, achieving self-sustaining enzymatic dynamic therapy (EDT)-photodynamic therapy (PDT) (EDT-PDT) synergy. The endogenous cascade-amplified EDT not only enhances the 1 O 2 efficacy in exogenous PDT therapy, but the intermediate O 2 can also alleviate local neuropathic pain caused by hypoxia for safe PDT treatment. This work pioneers enzyme-mediated dynamic control of tumor redox homeostasis, establishing a new therapeutic axis between biocatalytic amplification and photodynamic processes.
研究领域:
肿瘤学
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生物材料
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纳米医学
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分子生物学
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转化医学
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氧化还原生物学
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药物递送系统
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生物催化
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产品关联
DOI
PubMed
高影响因子
Advanced Fiber Materials
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2025
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IF 21.3
Zhao Qing,Li Yuping,Zhang Jinzheng,Lei Xiaoyu,Tang Jiajing,Chen Jieqiong,Li Jidong,Guo Weihua,Zuo Yi,Li Yubao
The rejuvenation of bone tissue remains a formidable challenge for osteoporosis (OP) patients who suffer severe bone degeneration or structural deterioration. To reverse the bone loss associated with OP, a dual-enzyme cascade system (PCF@EA) was engineered as a biomimetic engine to regulate bone metabolism imbalance. The bienzyme-driven system was constructed by integrating functionalized polymeric composite fibers with two mineralization-promoting hydrolases: recombinant human ectonucleotide pyrophosphatase/phosphodiesterase 1 (rhENPP1) and recombinant human alkaline phosphatase (rhALP). The bienzyme-driven engine efficaciously navigates mitochondria-activated mineralization through an autophagic process, thereby promoting osteogenic differentiation, increasing intracellular inorganic phosphate (Pi), and facilitating calcium influx. Concurrently, metabolic regulation mediated by the bienzyme-driven engine involves the PI3K–Akt pathway, tricarboxylic acid (TCA) cycle and glycerophosphate metabolism, significantly down-regulating the mineralization inhibitor pyrophosphate (PPi) while accelerating the formation of phosphate-related metabolites. Moreover, the enzyme-loaded substrates inhibited bone resorption by reducing expression of osteoclast activity markers, including Trap and Cath-K. This bienzymatic cascade strategy has demonstrated efficacy in restoring bone homeostasis in osteoporotic rats, significantly improving defect maturation and catalyzing the recovery of bone mineral density from severe loss back to baseline levels. The unique features of the bienzyme-driven engine provide a promising approach for the therapeutic treatment of degenerative skeletal diseases.
研究领域:
生物医学工程
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分子生物学
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生物化学
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再生医学
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药理学
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DOI
高影响因子
ANALYTICAL CHEMISTRY
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2025
·
IF 6.7
Jiaqi Xue,Kang Mao,Zihui Tang,Jiming Hu,Hua Zhang
Owing to the severe environmental and health issues posed by organophosphorus pesticides (OPs), a dual-enzyme cascade biosensing platform based on manganese dioxide (MnO2) and CRISPR/Cas12a was developed in this study. Smartphones were innovatively integrated with a stacked ensemble learning (SEL) model for ultrasensitive detection and dynamic monitoring of OPs in environmental water samples. A dual-enzyme cascade signal amplification strategy was used to construct this sensing platform. Acetylcholinesterase (AChE) catalyzes the generation of thiocholine (TCh), which, in turn, regulates the degradation of MnO2 nanosheets, releasing Cas12a activators and generating a fluorescence signal. Owing to the irreversible inhibition of AChE activity by OPs, dichlorvos (DDVP) was successfully detected, with a detection limit as low as 4.62 pg/mL. Additionally, the SEL model, integrated into the smartphone biosensing platform and incorporating random forest (RF), XGBoost, and ridge regression algorithms, exhibited strong performance in detecting OPs after optimization (R2 = 0.9985). In real water samples, the SEL model achieved a recovery rate of 93.1–103.1%, and the degradation kinetics of DDVP were successfully monitored over 24 h, revealing significant differences in DDVP degradation rates across various water matrices. This study is the first to report the integration of CRISPR/Cas12a biosensing technology with an SEL model-driven smartphone detection platform, providing a novel approach for sensitive, portable, and intelligent monitoring of OPs and offering new insights for water quality monitoring and early detection of environmental risks.
研究领域:
环境监测
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分析化学
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生物传感
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分子生物学
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环境毒理学
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水污染控制
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产品关联
DOI
PubMed
高影响因子
BIOTECHNOLOGY AND BIOENGINEERING
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2025
·
IF 3.6
Liang Li,Hui Jiang,Tingting Xu,Xi Wang
Citronellal and citronellol are monoterpenoids that serve as key ingredients in the flavor and fragrance industry and hold significant potential for pharmaceutical applications. Currently, these compounds are primarily obtained through chemical synthesis or extraction from plant essential oils. However, these methods raise concerns regarding environmental sustainability, carbon emissions, and the growing attentions for natural production. A promising alternative is microbial biosynthesis, which enables the natural and carbon-neutral production of these compounds in an environmentally friendly manner. In this study, we developed bienzymatic cascades for the efficient production of citronellal and citronellol from geraniol, utilizing E. coli 's endogenous glycerol dehydrogenase (GLDA), NADPH-dependent aldehyde reductase (AHR), and Catharanthus roseus iridoid synthase (CrIS). The highest in vitro production reached 92 mg/L of citronellal and 170 mg/L of citronellol in a 5-h enzymatic reaction. We also engineered these enzymatic cascades into E. coli strains for in vivo production, achieving up to 714 mg/L of citronellol. This study establishes new bienzymatic cascades for the efficient biosynthesis of citronellal and citronellol, demonstrating a possible bio-based production for natural flavor and fragrance compounds.
研究领域:
合成生物学
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生物催化
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代谢工程
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工业微生物学
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酶学
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产品关联
DOI
PubMed
高影响因子
Biotechnology for Biofuels and Bioproducts
·
2023
·Vol. 16
·期 1
·1-13
·
IF 0
Yang Fan,Liu Jiashu,Li Bianxia,Li Huanan,Jiang Zhengbing
Background As a cost-effective and eco-friendly approach, biocatalysis has great potential for the transformation of 5-hydroxymethylfurfural (HMF) into 2,5-furandicarboxylic acid (FDCA). However, the compatibility of each enzyme in the cascade reaction limits the transformation efficiency of HMF to FDCA. Results Coupled with an alcohol oxidase from Colletotrichum gloeosporioides ( Cgl AlcOx), this study aims to study the potential of bacterial laccase from Bacillus pumilus ( Bp Lac) in an enzymatic cascade for 2,5-furandicarboxylic acid (FDCA) biosynthesis from 5-hydroxymethylfurfural (HMF). Bp Lac showed 100% selectivity for HMF oxidation and generated 5-hydroxymethyl-2-furancarboxylic acid (HMFCA). Cgl AlcOx was capable of oxidizing HMFCA to 2-formyl-5-furancarboxylic acid (FFCA). Both Bp Lac and Cgl AlcOx could oxidize FFCA to FDCA. At the 5 mM scale, a complete transformation of HMF with a 97.5% yield of FDCA was achieved by coupling Bp Lac with Cgl AlcOx in the cascade reaction. The FDCA productivity in the reaction was 5.3 mg/L/h. Notably, Bp Lac could alleviate the inhibitory effect of FFCA on Cgl AlcOx activity and boost the transformation efficiency of HMF to FDCA. Moreover, the reaction was scaled up to 40 times the volume, and FDCA titer reached 2.6 mM with a yield of 58.77% at 168 h. Conclusions This work provides a candidate and novel insight for better design of an enzymatic cascade in FDCA production.
研究领域:
生物催化
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生物化学
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化学工程
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酶学
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DOI
PubMed
CATALYSIS LETTERS
·
2024
·
IF 2.3
Shen Nan,Hua Shaofeng
A dual-enzyme cascade catalytic system (Cu-rGO-Fe 3 O 4 -GA-CEL-PEI-GOD) was prepared by co-immobilizing cellulase (CEL) and glucose oxidase (GOD) on a nanocomposite (Cu-rGO-Fe 3 O 4 ) to efficiently catalyze the conversion of carboxymethyl cellulose (CMC) to gluconic acid. A layer-by-layer strategy was used by adding polyethyleneimine (PEI) to allow the upper enzyme layer to attach to the lower neighboring layer, increasing the loading capacity of the support. The loading capability of CEL and GOD on Cu-rGO-Fe 3 O 4 -GA-CEL-PEI-GOD were 55.034 mg g −1 and 12.4 mg g −1 , respectively. The specific activity of CEL on Cu-rGO-Fe 3 O 4 -GA-CEL was 74.3 U·g −1 , and that of immobilized CEL after cross-linking PEI was 25.45 U·g −1 , which could retain 34.253% of the enzyme activity. The specific activity of GOD on Cu-rGO-Fe 3 O 4 -GA-CEL-PEI-GOD was 30.9 U·g −1 . The carrier Cu-rGO-Fe 3 O 4 has peroxidase-like activity, which can timely remove harmful H 2 O 2 to the enzyme, thereby improving the yield of gluconic acid and the stability of biocatalysts. The yield of gluconic acid with Cu-rGO-Fe 3 O 4 -GA-CEL-PEI-GOD reached 96.04% within 2 h, higher than the control systems for comparison. In addition, the Cu-rGO-Fe 3 O 4 -GA-CEL-PEI-GOD maintained 82.61% of catalytic activity even after undergoing seven cycles of reaction. The dual-enzyme catalytic systems had shallow temperature and pH optima of 40 °C and 4.5. Such a chemoenzymatic cascade system provides a new strategy for the conversion from CMC to gluconic acid in one-step. Graphical Abstract Enzymatic conversion of sodium carboxymethyl cellulose (CMC-Na) to gluconic acid.
研究领域:
生物催化
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纳米技术
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酶工程
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材料科学
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产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）