Nanomaterials
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2021
·Vol. 11
·期 9
·2171
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IF 5.076
Le Wang,Pengxue Sun,Yiyu Yang,Hanzhen Qiao,Hailong Tian,Dapeng Wu,Shuoye Yang,Qipeng Yuan,Jinshui Wang
The field of enzyme cascades in limited microscale or nanoscale environments has undergone a quick growth and attracted increasing interests in the field of rapid development of systems chemistry. In this study, alcohol dehydrogenase (ADH), lactate dehydrogenase (LDH), and mesoporous silica nanoparticles (MSN) immobilized nicotinamide adenine dinucleotide (NAD+) were successfully immobilized on the zeolitic imidazolate frameworks (ZIFs). This immobilized product was named ZIF@ADH/NAD-MSN/LDH, and the effect of the multi-enzyme cascade was studied by measuring the catalytic synthesis of lactic acid. The loading efficiency of the enzyme in the in-situ co-immobilization method reached 92.65%. The synthesis rate of lactic acid was increased to 70.10%, which was about 2.82 times that of the free enzyme under the optimal conditions (40 °C, pH = 8). Additionally, ZIF@ADH/NAD-MSN/LDH had experimental stability (71.67% relative activity after four experiments) and storage stability (93.45% relative activity after three weeks of storage at 4 °C; 76.89% relative activity after incubation in acetonitrile-aqueous solution for 1 h; 27.42% relative activity after incubation in 15% N, N-Dimethylformamide (DMF) solution for 1 h). In summary, in this paper, the cyclic regeneration of coenzymes was achieved, and the reaction efficiency of the multi-enzyme biocatalytic cascade was improved due to the reduction of substrate diffusion.Keywords:confinement effect;multi-enzyme catalytic cascade;in-situ synthesis;coenzyme recycling;co-immobilization enzyme
研究领域:
生物化学
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纳米生物学
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系统化学
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DOI
PubMed
高影响因子
Nanomaterials
·
2018
·Vol. 8
·期 2
·127
·
IF 3.504
Sen Lin,Shiyong Sun,Ke Wang,Kexuan Shen,Biaobiao Ma,Yuquan Ren,Xiaoyu Fan
The bioinspired design and construction of enzyme@capsule microreactors with specific cell-like functionality has generated tremendous interest in recent years. Inspired by their fascinating complexity, scientists have endeavored to understand the essential aspects of a natural cell and create biomimicking microreactors so as to immobilize enzymes within the hierarchical structure of a microcapsule. In this study, simultaneous encapsulation of alcohol dehydrogenase (ADH) was achieved during the preparation of microcapsules by the Pickering emulsion method using amphiphilic modified TiO2nanoparticles (NPs) as building blocks for assembling the photocatalytic microcapsule membrane. The ADH@TiO2NP microreactors exhibited dual catalytic functions, i.e., spatially confined enzymatic catalysis and the membrane-associated photocatalytic oxidation under visible light. The sustainable cycling of nicotinamide adenine dinucleotide (NAD) coenzyme between NADH and NAD+was realized by enzymatic regeneration of NADH from NAD+reduction, and was provided in a form that enabled further photocatalytic oxidation to NAD+under visible light. This bioinspired ADH@TiO2NP microreactor allowed the linking of a semiconductor mineral-based inorganic photosystem to enzymatic reactions. This is a first step toward the realization of sustainable biological cycling of NAD+/NADH coenzyme in synthetic functional microsystems operating under visible light irradiation.Keywords:enzyme encapsulation;bioinspired microreactors;nicotinamide coenzymes cycling;alcohol dehydrogenase;TiO2nanoparticles
研究领域:
生物化学
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材料科学
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光催化
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DOI
PubMed
高影响因子
BIOTECHNOLOGY AND BIOENGINEERING
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2024
·
IF 3.5
Han-Yu Liu,Pan Ning,Feng Qian,Yao-Wu Wang,Hai-Min Zhang,Pu Wang
Flavin-dependent halogenase (FDH) is highly prized in pharmaceutical and chemical industries for its exceptional capacity to produce halogenated aromatic compounds with precise regioselectivity. This study has devised a multi-enzyme self-assembly strategy to construct an effective and reliable in vitro coenzyme cycling system tailored for FDHs. Initially, tri-enzyme self-assembling nanoclusters (TESNCs) were developed, comprising glucose dehydrogenase (GDH), flavin reductase (FR) and FDH. The TESNCs exhibited enhanced thermal stability and conversion efficiency compared to free triple enzyme mixtures during the conversion of L -Trp to 6-Cl- L -Trp, resulting in a 2.1-fold increase in yield. Subsequently, an ordered co-immobilization of GDH, FR, and FDH was established, further amplifying the stability and catalytic efficiency of the FDH coenzyme cycle system. Compared to the free TESNCs, the immobilized TESNCs demonstrated a 4.2-fold increase in catalytic efficiency in a 5 mL reaction system. This research provides an effective strategy for developing a robust and efficient coenzyme recycling system for FDHs.
研究领域:
生物化学
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合成生物学
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酶工程
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DOI
PubMed
高影响因子
Catalysts
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2025
·Vol. 15
·期 12
·
IF 4
Yang Yang,Kangkang Shang,Xiaorui Gao,Xingmiao Zhu,Mengying Ling,Pu Zheng,Shichao Xu,Pengcheng Chen
Raspberry ketone (RK) is the primary aromatic compound in raspberry fruit, which is widely utilized in perfume, cosmetics, and food additive industries. Currently, RK is predominantly produced chemically. RK biosynthesis through enzyme or whole cell has garnered significant attention due to the mild reaction conditions and the process being regarded as ‘natural’. This study proposed a ‘dual-microorganism, two-phase’ stepwise cascade strategy to produce RK from an economical precursor, 4-hydroxybenzaldehyde (4-HBD). An acetone-tolerant deoxyribose-phosphate aldolase DERAEc(S238D) mutant was obtained through a site-specific rigidification strategy for converting 4-HBD to 4-hydroxybenzylaceton (4-HBA). Then, an engineeredE. colico-expressing isocitrate dehydrogenase and raspberry ketone synthaseRiRZS1 with a citrate-sodium citrate buffer to recycle nicotinamide adenine dinucleotide phosphate (NADPH) was constructed for the conversion of 4-HBA to RK. The final concentration of RK was 50.00 ± 1.92 mmol·L−1with a yield of 86.96%. This strategy provides a scalable coenzyme self-recycling and two-phase catalysis platform for high-value phenolic compounds.
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高影响因子
Bioresources and Bioprocessing
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2022
·Vol. 9
·期 1
·1-9
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IF 4.983
Ma Yue,Li Tianzhen,Tan Zijian,Ma Long,Liu Haifeng,Zhu Leilei
Catalytic valorization of raw glycerol derived from biodiesel into high-value chemicals has attracted great attention. Here, we report chemoenzymatic cascade reactions that convert glycerol to lactic acid and glycolic acid. In the enzymatic step, a coenzyme recycling system was developed to convert glycerol into 1,3-dihydroxyacetone (DHA) with a yield of 92.3% in potassium phosphate buffer (300 mM, pH 7.1) containing 100 mM glycerol, 2 mM NAD + , 242 U/mL glycerol dehydrogenase-GldA and NADH oxidase-SpNox K184R at 30 °C. Subsequently, NaOH or NaClO 2 catalyzes the formation of lactic acid and glycolic acid from DHA. The high yield of lactic acid (72.3%) and glycolic acid (78.2%) verify the benefit of the chemoenzymatic approaches. Graphical Abstract
研究领域:
化学工程
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生物技术
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催化
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生化
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DOI
高影响因子
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