BIOPROCESS AND BIOSYSTEMS ENGINEERING
·
2016
·Vol. 39
·期 4
·603-611
·
IF 1.901
Zhang Jian-Dong,Cui Zhi-Mei,Fan Xiao-Jun,Wu Hua-Lei,Chang Hong-Hong
Two uncharacterized nicotinamide adenine dinucleotide (NADH) oxidases (named as LpNox1, LpNox2) from Lactobacillus pentosus ATCC 8041 were cloned and overexpressed in Escherichia coli BL21 (DE3). The sequence analysis revealed that the two enzymes are water-forming Noxs with 64 % and 52 % identity to LbNox from Lactobacillus brevis DSM 20054. The optimal pH and temperature of the purified LpNox1 and LpNox2 were 7.0 and 8.0 and 35 and 40 °C, respectively, with K
M of 99.0 μM (LpNox1) and 27.6 μM (LpNox2), and yielding catalytic efficiency k
cat / K
M of 1.0 and 0.2 μM −1 s −1 , respectively. Heat inactivation studies revealed that the two enzymes are relatively instable. The application of LpNox1 for the regeneration of NAD + was demonstrated by coupling with a glycerol dehydrogenase-catalyzed oxidation of glycerol to 1,3-dihydroxyacetone. The characteristics of the LpNox1 could prove to be of interest in industrial application such as NAD + regeneration in dehydrogenase-catalyzed oxidations.
研究领域:
生物化学
·
微生物学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2019
·
IF 4.784
Meng-Qiu Xu,Fei-Long Li,Wen-Qian Yu,Rui-Fang Li,Ye-Wang Zhang
Cofactor regeneration is an important method to avoid the consumption of large quantities of oxidized cofactor NAD + in enzyme-catalyzed reactions. Herein, glycerol dehydrogenase (GDH) and NADH oxidase preparations by aggregating enzymes with ammonium sulphate followed by cross-linking formed aggregates for effective regeneration of NAD + . After optimization, the activity of combi-CLEAs and separate CLEAs mixtures were 950 and 580 U/g, respectively. And the catalytic stability of combi-CLEAs against pH and temperature was superior to the free enzyme mixture. After ten cycles of reuse, the catalytic efficiency could still retain 63.3% of its initial activity, indicating that the constructed combi-CLEAs system had excellent reusability. Also, the conversion of glycerol to 1,3-dihydroxyacetone (DHA) was improved by the constructed NAD + regeneration system, resulting in 4.6%, which was 2.5 times of the free enzyme system. Thus, wide applications of this co-immobilization method in the production of various chiral chemicals could be expected in the industry for its high efficiency at a low cost.
研究领域:
生物化学
·
酶工程
✓ 已验证
DOI
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2019
·
IF 4.784
Fei-Long Li,Qiang Zhou,Wei Wei,Jian Gao,Ye-Wang Zhang
Enzymatic NADP + regeneration is a promising approach to produce valuable chemicals under economic conditions. Among all the enzymatic routes, using water-forming NADH oxidase is an ideal one because there is no by-product. However, most NADH oxidases have a low specific activity to NADPH. In this work, a thermostable NADH oxidase from Lactobacillus rhamnosus (LrNox) was rationally engineered to switch its specificity from NADH to NADPH. The results show that mutants D177A, G178R, D177A/G178R, D177A/G178R/L179S improved the NADPH activity by a factor of 4–6. The highest NADPH catalytic efficiency ( K cat /K m 223.71 S −1 μm −1 , 47.6-fold higher than wild-type LrNox) and 51% of NADH activity retention were achieved by replacing the single amino acid Leu179 for serine (L179S) in LrNox. Modeling of L179S-NADPH complex reveals that the phosphate group of NADPH interacts with the hydroxyl of Ser179 with a strong hydrogen bond and several shorter hydrogen bonds with the amino group of Lys185 could stabilize the binding of NADPH in the L179S mutant. This work provides an efficient method for converting NAD(P)H specificity and shows that L179S mutant is a potential and efficient auxiliary enzyme for NADP + regeneration.
研究领域:
生物化学
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生物技术
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酶学
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产品关联
DOI
PubMed
高影响因子
Energies
·
2022
·Vol. 15
·期 12
·4242
·
IF 3.252
Xue-Yong Li,Jia-Ying Huang,Qiang Zhou,Yuan-Yuan Xu,Ponnandy Prabhu,Ye-Wang Zhang
Acetoin, a four-carbon hydroxyl-keto compound, is used in the food, pharmaceutical, and chemical industries. The cascade enzymatic production is considered a promising and efficient method to produce acetoin. However, the stability and compatibility of the enzymes under the same catalytic conditions are challenges that need to be resolved. In this work, alcohol dehydrogenase, acetaldehyde lyase, and NADH oxidase were selected to work at the same conditions to efficiently convert ethanol into acetoin. These three enzymes were immobilized on epoxy-modified magnetic nanomaterials to obtain highly stable biocatalysts. The stability and the immobilization conditions, including temperature, pH, enzyme–carrier ratio, and immobilization time, were optimized to obtain the immobilized enzymes with a high catalytic activity. The cascade reactions catalyzed by the immobilized enzymes yielded a high conversion of 90%, suggesting that the use of immobilized enzymes is a promising way to produce acetoin.Keywords:immobilization;NADH oxidase;acetaldehyde lyase;alcohol dehydrogenase;acetoin;cascade enzymatic reactions
研究领域:
生物技术
·
酶学
·
化学工程
✓ 已验证
产品关联
DOI
高影响因子
RARE METALS
·
2025
·
IF 11
Xue Hong-Jin,Zhang Meng,Li Yong-Qing,Liu Chao-Ran,Ma Xin-Yu,He Qin-Bin,Wang Yin-Chuan,Chen Jin-Xing,Qiu Jian-Feng
Constructing high-performance nanozymes for specific biomolecules is crucial but challenging for practical applications and fundamental research. Herein, through the examination of the catalytic reaction paths of natural nicotinamide adenine dinucleotide (NADH) oxidase (NOX), a novel and efficient single-atom rhodium catalyst (Rh 1 /NC) was developed to mimic NOX. The Rh 1 /NC demonstrated the ability to catalyze the dehydrogenation of NADH and transfer electrons to O 2 to generate H 2 O 2 through the typical two-electron pathway. Furthermore, our findings revealed that Rh 1 /NC exhibits the ability to catalyze the conversion of produced H 2 O 2 into OH under mildly acidic conditions. This process amplifies the oxidation of NADH, showcasing NADH peroxidase-like activity (NPx-like). As a paradigm, this unique dual enzyme-like property of Rh 1 /NC with a positive feedback effect holds significance in disrupting cancer cellular homeostasis. Rh 1 /NC can effectively consume NADH via cascade biocatalytic reactions within cancer cells, further triggering the elevation of reactive oxygen species (ROS), leading to impaired oxidative phosphorylation and decreased mitochondrial membrane potential, thus damaging the adenosine triphosphate (ATP) synthesis. The resulting 'domino effect' interferes with the energy metabolism homeostasis of cancer cells, ultimately promoting cell apoptosis. This study provides potential guidance for the rational design of materials with greater capabilities. Graphical abstract
研究领域:
生物化学
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纳米医学
·
肿瘤学
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY
·
2018
·Vol. 93
·期 8
·2351-2358
·
IF 2.587
Meng-Yao Zhuang,Xiao-Ping Jiang,Xiao-Min Ling,Meng-Qiu Xu,Yi-Hao Zhu,Ye-Wang Zhang
BACKGROUND Nicotinamide cofactor-dependent oxidoreductases have been widely applied to the bioproduction of varieties of useful compounds. Efficient cofactor regeneration is often required for these oxidoreductase-catalyzed reactions. Herein, enzymatic production of 1, 3-dihydroxyacetone (DHA) via immobilized enzymes with in situ cofactor regeneration was reported. RESULTS Glycerol dehydrogenase (GDH) and NADH oxidase (Nox) were immobilized on two carriers: macroporous resin and epoxy functionalized magnetic nanoparticles (EFMN). Higher enzyme loading capacity and activity recovery were achieved on EFMN. A high immobilization yield of 90% and >70% recovery activity were obtained for both enzymes using EFMN as carriers. Besides, both immobilized enzymes showed enhanced pH stability and temperature tolerance. Co-immobilized and mixed immobilized enzymes were evaluated for synthesis of DHA, and mixed immobilized enzymes showed higher catalytic rate. The mixed reaction system also was investigated in view of various operational factors. Under the optimal conditions, a high DHA concentration with mixed immobilized enzymes system can reach up to 3.5 mmol L –1 , which was about four times higher than that without cofactor regeneration system. CONCLUSION The immobilized GDH and Nox on magnetic nanoparticles showed enhanced stabilities. A cofactor regeneration system with immobilized enzymes could be utilized for efficient production of DHA from inexpensive substrates. © 2018 Society of Chemical Industry
研究领域:
生物化学
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酶工程
✓ 已验证
产品关联
DOI
高影响因子
New Biotechnology
·
2021
·
IF 5.079
Wen-Bin Su,Fei-Long Li,Xue-Yong Li,Xiao-Man Fan,Rui-Jiang Liu,Ye-Wang Zhang
L-Tagatose, a promising building block in the production of many value-added chemicals, is generally produced by chemical routes with a low yield, which may not meet the increasing demands. Synthesis of l -tagatose by enzymatic oxidation of d -galactitol has not been applied on an industrial scale because of the high cofactor costs and the lack of efficient cofactor regeneration methods. In this work, an efficient and environmentally friendly enzymatic method containing a galactitol dehydrogenase for d -galactitol oxidation and a water-forming NADH oxidase for regeneration of NAD + was first designed and used for l -tagatose production. Supplied with only 3 mM NAD + , subsequent reaction optimization facilitated the efficient transformation of 100 mM of d -galactitol into l -tagatose with a yield of 90.2 % after 12 h (obtained productivity: 7.61 mM.h −1 ). Compared with the current chemical and biocatalytic methods, the strategy developed avoids by-product formation and achieves the highest yield of l -tagatose with low costs. It is expected to become a cleaner and more promising route for industrial biosynthesis of l -tagatose.
研究领域:
生物技术
·
生物化学
·
酶学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOPROCESS AND BIOSYSTEMS ENGINEERING
·
2021
·Vol. 45
·期 2
·353-364
·
IF 3.21
Li Xue-Yong,Xu Meng-Qiu,Liu Hui,Zhou Qiang,Gao Jian,Zhang Ye-Wang
The combined cross-linked enzyme aggregates (combi-CLEAs) containing galactitol dehydrogenase (Gdh) and NADH oxidase (Nox) were prepared for l -tagatose synthesis. To prevent the excess consumption of cofactor, Nox in the combi-CLEAs was used to in situ regenerate NAD + . In the immobilization process, ammonia sulfate and glutaraldehyde were used as the precipitant and cross-linking reagent, respectively. The preparation conditions were optimized as follows: 60% ammonium sulfate, 1:1 (molar ratio) of Gdh to Nox, 20:1 (molar ratio) of protein to glutaraldehyde, and 6 h of cross-linking time at 35 °C. Under these conditions, the activity of the combi-CLEAs was 210 U g −1 . The combi-CLEAs exhibited higher thermostability and preserved 51.5% of the original activity after eight cycles of reuses at 45 °C. The combi-CLEAs were utilized for the preparation of l -tagatose without by-products. Therefore, the combi-CLEAs have the industrial potential for the bioconversion of galactitol to l -tagatose. Graphical abstract
研究领域:
生物技术
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酶学
·
化学工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Inorganic Chemistry Frontiers
·
2024
·
IF 6.1
Xuwen Da,Yunli Xu,Lei Wang,Xiulian Liu,Yatong Peng,Yao Wu,Wanpeng Zhou,Wentao Wang,Xuesong Wang,Qian-Xiong Zhou
Traditional photodynamic therapy (PDT) is almost ineffective against deeply seated tumors due to the limited penetration depth of external light. The development of chemiexcited PDT is promising to address this issue, but it is still challenging to construct simple, efficient and targeted chemiexcited PDT systems. In this work, CPPO (bio(3,5,6-trichloro-2-n-pentyloxycarbonylphenyl) oxalate) and [Ru(phen)2(PIP-OCH3)](PF6)2 (Ru3, where phen = 1,10-phenanthroline, PIP-OCH3 = 2-(4-methoxy phenyl)-1Himidazo[4,5-f][1,10]phenanthroline) with efficient photo-catalytic NADH oxidation were co-encapsulated with DSPE-PEG 2000 to form Ru3/CPPO@PEG nanoparticles (NP3). Once internalized by cancer cells, Ru3 can be excited by the chemical energy produced by the reaction between CPPO and endogenous H2O2. The subsequent oxidation of intracellular NADH and generation of H2O2 in situ by excited Ru3 will further promote the sustained occurrence of chemical reaction, realizing a self-enhanced chemiexcited PDT. NP3 could effectively deplete intracellular NADH, inhibit ATP synthesis, increase H2O2 content, and produce reactive oxygen species (ROS) within cancer cells, thus exhibited efficient chemiexcited PDT activity against various cancer cells. Notably, minimal cytotoxicity was observed towards normal cells. Good therapeutic effects against subcutaneously implanted and lung metastatic tumors were also observed for NP3. The current findings may provide new sights for development of novel chemiexcited PDT.
研究领域:
光动力疗法
·
癌症研究
·
纳米技术
·
生物化学
·
药学
✓ 已验证
产品关联
DOI
高影响因子
Inorganic Chemistry Frontiers
·
2020
·Vol. 7
·期 20
·4020-4027
·
IF 5.958
Chen Ge,Jiayi Zhu,Ai Ouyang,Nong Lu,Yi Wang,Qianling Zhang,Pingyu Zhang
Osmium(II) complexes usually exhibit an excellent NIR (near-infrared) emission, which conforms to the optical window in a biological system. Here we designed three mitochondria-targeted NIR terpyridine Os(II) complexes as photosensitizers, which demonstrated great NIR phosphorescence emission and long phosphorescence lifetime. The emission spectra of the osmium(II) complexes were red-shifted with increasing conjugated planes of their terpyridine ligand (Os3 > Os2 > Os1). Furthermore, the Os(II) complexes, especially Os3, could produce singlet oxygen and oxidize NADH under both 465 nm and 633 nm LED light irradiation. It was found that Os3 has certain phototoxicities upon red light irradiation, and have great potential for NIR photodynamic therapy.
研究领域:
光动力疗法
·
光化学
·
无机化学
·
生物医学工程
✓ 已验证
产品关联
DOI
高影响因子
GREEN CHEMISTRY
·
2018
·Vol. 20
·期 10
·2382-2391
·
IF 8.586
Feng Sha,Yucong Zheng,Jiao Chen,Kequan Chen,Fei Cao,Ming Yan,Pingkai Ouyang
In this study, the bio-oxidation of galactitol to the valuable D-tagatose is presented. This proposed strategy could start with the refined by-product of waste xylose mother liquor formed by chemical hydrogenation. Through a computationally guided enzyme screening approach, a robust polyol dehydrogenase (PdPDH) was rapidly identified from a massive number of candidates. When coupled with a water-forming NADH oxidase (StNOX) for environmentally benign cofactor regeneration, subsequent reaction optimization facilitated the complete transformation of 100 g L−1 galactitol into the desired product within 15 h, with a yield of 91% in a 2 L scale preparative reaction. Compared to the current enzymatic isomerization system, our approach avoids low conversion, high operative temperatures and by-product formation, while enabling simplified product isolation.
研究领域:
生物技术
·
酶学
·
可持续化学
✓ 已验证
产品关联
DOI
高影响因子
BMC Veterinary Research
·
2022
·Vol. 18
·期 1
·1-14
·
IF 2.792
Hao Fei,Xie Xing,Feng Zhixin,Chen Rong,Wei Yanna,Liu Jin,Xiong Qiyan,Shao Guoqing,Lin Johnson
Background Mycoplasma hyopneumoniae (M. hyopneumoniae) is the etiological agent of enzootic pneumonia, a highly infectious swine respiratory disease that distributed worldwide. The pathogenesis and virulence factors of M. hyopneumoniae are not fully clarified. As an important virulence factor of bacteria, nicotinamide adenine dinucleotide (NADH) oxidase (NOX) participates in host-pathogen interaction, however, the function of NOX involved in the pathogenesis of M. hyopneumoniae is not clear.Results In this study, significant differences in NOX transcription expression levels among different strains of M. hyopneumoniae differed in virulence were identified, suggesting that NOX may be correlated with M. hyopneumoniae virulence. The nox gene of M. hyopneumoniae was cloned and expressed in Escherichia coli, and polyclonal antibodies against recombinant NOX (rNOX) were prepared. We confirmed the enzymatic activity of rNOX based on its capacity to oxidize NADH to NAD+. Flow cytometry analysis demonstrated the surface localization of NOX, and subcellular localization analysis further demonstrated that NOX exists in both the cytoplasm and cell membrane. rNOX was depicted to mediate adhesion to immortalized porcine bronchial epithelial cells (hTERT-PBECs). Pre-neutralizing M. hyopneumoniae with anti-rNOX antibody resulted in a more than 55% reduction in the adhesion rate of high- and low-virulence M. hyopneumoniae strains to hTERT-PBECs. Moreover, a significant difference appeared in the decline in CCU50 titer between virulent (168) and virulence-attenuated (168L) strains. NOX not only recognized and interacted with host fibronectin but also induced cellular oxidative stress and apoptosis in hTERT-PBECs. The release of lactate dehydrogenase by NOX in hTERT-PBECs was positively correlated with the virulence of M. hyopneumoniae strains.Conclusion sNOX is considered to be a potential virulence factor of M. hyopneumoniae and may play a significant role in mediating its pathogenesis.
研究领域:
微生物学
·
兽医学
·
分子生物学
·
病理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）