GREEN CHEMISTRY
·
2026
·
IF 10.6
Hong-Ru Zhao,Wen-Tao Shen,Bingmei Su,Lian Xu,Juan Lin
(1R,2R)-2-Amino-1-(4-(methylsulfonyl)phenyl)propane-1,3-diol [(1R,2R)-3] is a key chiral intermediate for the antibiotic florfenicol. Its asymmetric synthesis remains challenging due to the difficulty in constructing two contiguous stereocenters with high selectivity. Herein, we report a green and sustainable one-pot enzymatic cascade for the efficient synthesis of (1R,2R)-3 from inexpensive and readily available L/D-serine and 4-(methylsulfonyl)benzaldehyde (4-Mb). The cascade starts with the L-amino acid oxidase (AncLAAO)-mediated oxidation of L-serine to produce β-hydroxypyruvate (1). The subsequent pyruvate decarboxylase (SwPDC)-catalyzed decarboxylative condensation led to the asymmetric assembly of 1 and 4-Mb to yield (R)-1,3-dihydroxy-1-(4-(methylsulfonyl)phenyl)propan-2-one [(R)-2], and an engineered transaminase (ATA117) aminates (R)-2 using D-serine as the amine donor to produce the target product (1R,2R)-3. This design inherently enables a one-pot cascade for assess (1R,2R)-3 and prevents the accumulation of inhibitory pyruvate, embodying the principles of atom economy and waste prevention. The rate-limiting transaminase ATA117 was engineered through a multi-strategy approach. The resulting optimal variant, ATA117-Mu9 demonstrated a 39-fold increase in catalytic efficiency (kcat/Km) and a 4.65-fold improvement in (1R,2R)-3 yield (27.9 mM) over the wild-type enzyme while maintaining a high diastereomeric excess (de) of 96-98% throughout the engineering process. For scalable application, a whole-cell biocatalyst BL21(01) co-expressing ancLAAO, SwPDC and ATA117-M9 was constructed. In a 100 mL scale-up reaction, 50 mM of 4-Mb was completely converted to 47.9 mM of (1R,2R)-3 within 3 h, achieving a space-time yield of 3.91 g/L/h, the highest reported for this compound. In summary, this work establishes a simple, efficient, and environmentally benign route that eliminates heavy metals, minimizes waste, and uses benign reaction media, providing a promising green alternative for the industrial synthesis of florfenicol intermediates.
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Engineering
·
2026
·
IF 12.2
Yiwen Shen,Ziqi Liang,Lanxue Li,Wei Zhang,Yaru Wang,Jian Tian,Bin Yao,Huoqing Huang,Huiying Luo,Tao Tu
Starch is a vital global commodity for food and industry, yet its agricultural production is unsustainable, relying on inefficient photosynthesis, arable land, and stable climates. Meanwhile, the annual production of ∼180 million tons of cellulose primarily from crop residues is largely wasted through incineration or landfilling. While enzymatic saccharification efficiently depolymerizes cellulose into glucose, its conversion to starch is fundamentally limited by the insufficient catalytic efficiency, poor substrate selectivity of key enzymes, and the reversibility of the core reactions they catalyze. Here, we optimized a cell-free three-enzyme cascade, designated the carbon-economical starch synthesis (CESS) pathway, to address this limitation. This in vitro cascade integrates three core enzymes: polyphosphate glucokinase (PPGK) for adenosine triphosphate (ATP)-free glucose activation, phosphoglucomutase (PGM), and α-glucan phosphorylase (αGP). We engineered the rate-limiting αGP from Thermotoga petrophila to generate the M7 variant ( Tp αGPM7), achieving a 4.54-fold higher catalytic efficiency, and optimized cofactors, identifying 20 mmol∙L −1 Mn 2+ to critically shift PGM’s preference toward starch synthesis. The optimized pathway converts 5.4 g∙L −1 glucose (equivalent to 30 mmol∙L −1 ) to 3.50 g∙L −1 starch at 60 ℃ with a record-breaking 71.93% conversion yield and a space-time yield of 2.33 g∙L −1 ∙h −1 , while minimizing carbon loss < 30%. The product was characterized as amylose. This photosynthesis-independent framework advances a sustainable route to valorize cellulosic waste into food-grade starch, offering a resilient strategy for food security and a circular bioeconomy.
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MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Mengjie Lin,Wenxin Chen,Cunshan Zhang,Jiewei He,Lina Wu,Ailin Liu,Xinhua Lin,Liqing Lin
Specific-Species enzyme detection serves as a crucial method in foodborne pathogen surveillance. Particularly, endogenous β-galactosidase (β-Gal) enzyme has been widely applied to determine the concentration of Escherichia coli ( E. coli ). The main challenges include reducing analysis time and improving detection sensitivity. A novel strategy for biosensing β-Gal activity and E. coli are developed based on BSA‑manganese dioxide nanosheets (BSA-MnO 2 NS) with the oxidase-mimicking activity to regulate the fluorescence of thiamine. BSA-MnO 2 NS can catalyze the oxidation of non-fluorescent thiamine to generate strong fluorescent dehydrothiamine. When β-Gal enzymes is introduced, p -aminophenyl-β-D-galactopyranoside (PAPG) could be hydrolyzed to form para-aminophenol (PAG), which efficiently reduced BSA-MnO 2 NS into Mn 2+ , resulting in the loss of their oxidase-like activity and the decrease of dehydrothiamine fluorescence. Since endogenous β-Gal in E. coli cells can directly hydrolyze PAPG without destroying bacteria, which greatly reduced the analysis time. The reduction product PAG leads to MnO 2 NS decomposition and thus reduced dehydrothiamine fluorescence. According to this mechanism, fluorescence biosensor for β-Gal activity and E. coli are constructed utilizing thiamine luminescence modulated by BSA-MnO 2 NS, without requiring an additional oxidizer. β-Gal and E. coli showed good linearity in the range of 0.01–2.0 μg/mL and 1.0 × 10 2 –1.0 × 10 4 CFU/mL, and the detection limits reached 8.55 ng/mL and 89.45 CFU/mL, respectively. The proposed enzyme-nanozyme cascade reaction significantly improved the sensitivity. It is highly specific and can be used to distinguish E. coli from other pathogenic bacteria. The strategy has also been successfully applied to the detection of E. coli in real samples.
研究领域:
生物传感
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微生物学
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纳米生物学
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分子诊断
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食品安全检测
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酶学
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Science Immunology
·
2025
·Vol. 10
·期 114
·
IF 16.3
Zhi Li,Jing Wang,Juan Du,Jun Li,Yanan Song,Junji Zhu,Ziyi Li,Wei Zhou,Shiyun Chen,Lijie Yang,Maohui Feng,Xiaolian Cai,Katryn J. Stacey,Wuhan Xiao
Processes that control tissue inflammation are essential to restore homeostasis after infection. The transcription factor NF-κB plays a central role in coordinating inflammation, but the mechanisms that regulate NF-κB signaling are not fully understood. Here, we identify OCEL1 ( occludin/ELL domain containing 1 ) as a negative regulator of NF-κB signaling. In the absence of infection, human OCEL1 bound to the LZ domain of NEMO (NF-κB essential modulator) and inhibited TRAF6-mediated K63-linked polyubiquitination, suppressing NF-κB signaling. During infection, OCEL1-mediated negative regulation of NF-κB signaling was impaired by FK506-binding protein bacterial peptidyl-prolyl cis/trans isomerases, which bound to OCEL1 in an amino-terminal palindromic proline–rich element (PPE) and promoted its degradation. Mice expressing a mutant version of the human OCEL1 PPE had dampened inflammation and increased susceptibility to Pseudomonas aeruginosa infection. Thus, the PPE of human OCEL1 senses bacterial infection, and its degradation releases the suppression of NF-κB signaling and promotes inflammation.
研究领域:
免疫学
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分子生物学
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传染病学
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信号转导
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2025
·
IF 6.2
Xiao-Ting Zhou,Ke-Yan Wei,Shan Li,Qi Shen,Ya-Ping Xue,Yu-Guo Zheng
Nitrilases provide a sustainable route for directly converting nitriles to carboxylic acids. However, efficient biocatalytic synthesis of the agrochemical intermediate 3,6-dichloropicolinic acid(3,6-DCPA) from 3,6-dichloropicolinonitrile (3,6-DCPN) remains unreported. We identified a novel nitrilase from Rhodococcus pyridinivorans (RpNIT) and developed the integrated Feature Cluster-Multi-Force field-Evolutionary Co-Localization (FMEC) strategy to simultaneously enhance activity, thermostability, and specificity. FMEC employs hierarchical clustering, molecular dynamics simulations, and multiforce field algorithms, identifying 19 candidate residues. Using Parallel Iterative Saturation Mutagenesis, we generated the quintuple mutant M5a-DGPAT. This variant exhibited a 17.2 °C increase in melting temperature and a 3.3 × 104-fold improvement in hydrolytic efficiency (kcat/Km) over wild-type RpNIT, while abolishing hydratase activity. Structural analysis attributed the superior performance to an optimized rigidity-flexibility balance, refined substrate binding pocket geometry, and strengthened interaction networks. M5a-DGPAT achieved complete hydrolysis of 347.9 mM 3,6-DCPN to 3,6-DCPA under mild conditions, demonstrating significant industrial potential. This study establishes a robust computational framework for designing highly active, stable, and specific engineered enzymes.
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MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Jie Cui,Yuxin Xia,Leilei Jin,Jishuang Chen,Zongli Huo,Fei Shen,Li Mi
Metal ions are widely present in biological systems and participate in many critical biochemical processes. Resisting interfering ions and achieving specific detection at the biosensor present a significant challenge. Herein, we designed a highly sensitive and anti-interference Bio-QDs biosensor through the biosynthesis CdSe quantum dots (QDs) and the use of sodium hexametaphosphate (HMP) as a stabilizer (CdSe QD@HMP) for detecting the calcium ion (Ca 2+ ) in plant tissue culture medium, which is known for its complexity. Because of the spatial hexahedron molecular structure and six interconnected tetrahedral P-O units of HMP increased the electronegativity and stability of CdSe, the CdSe QD@HMP easily captured Ca 2+ in solution, resulting in a minimum detection limit of 0.377 µM for Ca 2+ in the range of 1–300 μM. And HMP can also ensure that the detection process is not affected by negative ion interference. Additionally, the recovery rate of environmental samples was 96.40 % and 101.13 %, confirming the reliability and practicality of the prepared biosensors. Therefore, this strategy not only provides a green and safe pathway for synthesizing QDs, but also lays the foundation for the sensitive determination of metal cations in complex environment.
研究领域:
生物传感器技术
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植物学
·
分析化学
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高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2020
·
IF 5.162
Xiaoxiang Hu,Yajun Bai,Tai-Ping Fan,Xiaohui Zheng,Yujie Cai
A novel alanine dehydrogenase (ADH; EC.1.4.1.1) with high pyruvate reduced activity was isolated from Helicobacter aurati and expressed in Escherichia coli BL21 (DE3). The optimum pH of the reduction and oxidation reaction were 8.0 and 9.0, respectively, and the optimum temperature was 55 °C. With pyruvate and alanine as substrates, the specific activity of HAADH1 were 268 U·mg −1 and 26 U·mg −1 , respectively. HAADH1 had a prominent substrate specificity for alanine ( K m = 2.23 mM, k cat / K m = 8.1 s −1 ·mM −1 ). In the reduction reaction, HAADH1 showed the highest substrate affinity for pyruvate ( K m = 0.56 mM, k cat / K m = 364 s −1 ·mM −1 ). Compared to pyruvate, oxaloacetic acid, 2-ketobutyric acid, 3-fluoropyruvate, α-ketoglutaric acids, glyoxylic acid showed a residual activity of 93.30%, 8.93%, 5.62%, 2.57%, 2.51%, respectively. Phylogenetic tree analysis showed that this is a new type of ADH which have a low sequence similarity to available ADH reported in references. 3-Fluoropyruvate was effectively reduced to 3-fluoro-L-alanine by whole-cell catalysis.
研究领域:
生物化学
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微生物学
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酶学
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APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2020
·Vol. 104
·期 23
·10005-10018
·
IF 3.53
Guo Xiaolei,Wu Weibin,Zhang Mingliang,Wu Licheng,Huang Jianzhong
Abstract l -Tyrosine is a versatile compound used in the fine chemical, pharmaceutical, and functional food industries. Here, we report a bi-enzymatic cascade involving alanine racemase (ALR) and d -amino acid oxidase (DAAO) to produce pyruvate, as co-substrate for l -tyrosine production, from the cheap substrate l -alanine. The BpALR (ALR from Bacillus pseudofirmus ) was used as a whole-cell biocatalyst, converting l -alanine to d , l -alanine. The FsDAAO (DAAO from Fusarium solani ) was immobilized to oxidize the d -alanine generated in the first step to pyruvate. Both systems were combined as a continuous-flow reactor for maximized l -alanine-to-pyruvate conversion rates. The optimal parameters and appropriate conditions for FsDAAO immobilization were investigated. The pyruvate concentration of 86.6 g/L was achieved within 17 h. Subsequently, a whole-cell biocatalyst system for l -tyrosine production, catalyzed by the tyrosine phenol-lyase (TPL) from Erwinia herbicola (EhTPL), was developed, and a fed-batch approach was applied with phenol and the pyruvate produced with the ALR/DAAO system mentioned above. The concentration of phenol and pyruvate in the reactor should not exceed 7.5 g/L and 10 g/L, respectively. Significantly, the l -tyrosine concentration of 152.5 g/L was achieved within 10 h, demonstrating the great potential for high-efficiency production of l -tyrosine through the approach we established in this paper. Graphical abstract Key points • A specific bioreactor system for pyruvate produced from l -alanine was developed • The appropriate condition for immobilization of FsDAAO was investigated • A fed-batch process was established to produce l -tyrosine with recombinant E. coli • The bi-enzymatic cascade was successfully used for l -tyrosine production at low cost
研究领域:
生物技术
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生物化学
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化工工程
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药学
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JOURNAL OF HAZARDOUS MATERIALS
·
2024
·
IF 12.2
Lizhu Aer,Qifa Jiang,Linling Zhong,Qiuyue Si,Xianghong Liu,Yan Pan,Juan Feng,Hongjuan Zeng,Lixia Tang
Although many efforts have been devoted to the modification of polyethylene terephthalate (PET) hydrolases for improving the efficiency of PET degradation, the catalytic performance of these enzymes at near-ambient temperatures remains a challenge. Herein, a multi-enzyme cascade system (PT-EC) was developed and validated by assembling three well-developed PETases, PETase EHA , Fast-PETase, and Z1-PETase, respectively, together with carboxylesterase TfCa, and hydrophobic binding module CBM3a using scaffold proteins . The resulting PT-EC EHA , PT-EC FPE , PT-EC ZPE all demonstrated outstanding PET degradation efficacy. Notably, PT-EC EHA exhibited a 16.5-fold increase in product release compared to PETase EHA , and PT-EC ZPE yielded the highest amount of product. Subsequently, PT-ECs were displayed on the surface of Escherichia coli , respectively, and their degradation efficiency toward three PET types was investigated. The displayed PT-EC EHA exhibited a 20-fold increase in degradation efficiency with PET film compared to the surface-displayed PETase EHA . Remarkably, an almost linear increase in product release was observed for the displayed PT-EC ZPE over a one-week degradation period, reaching 11.56 ± 0.64 mM after 7 days. TfCa I69W/L281Y evolved using a docking-based virtual screening strategy showed a further 2.5-fold increase in the product release of PET degradation. Collectively, these advantages of PT-EC demonstrated the potential of a multi-enzyme cascade system for PET bio-cycling.
研究领域:
生物化学
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环境科学
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合成生物学
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JOURNAL OF HAZARDOUS MATERIALS
·
2020
·
IF 9.038
Ning Hou,Qiaoruo Wang,Yang Sun,Xianyue Li,Qiuying Song,Xinxin Jiang,Baoxin Li,Xinyue Zhao,Hailian Zang,Dapeng Li,Chunyan Li
In recent years, special attention has been devoted to biodemulsifiers as a new type of environment-friendly demulsifiers. A novel biodemulsifying oxalate decarboxylase (OxdC) secreted by Bacillus mojavensis XH1 is reported in the present study. A genome-wide comparison showed that strains with high demulsification efficiencies all possess alkane degradation genes. An analysis of the differentially expressed genes and proteins induced by different substrates showed that OxdC secreted by XH1 was an effective demulsifier. Moreover, the demulsification ability was verified by prokaryotic gene expression, knockout and complementation analyses. OxdC from XH1 exhibited a strong demulsification capacity and significantly outperformed the model protein Bacillus subtilis 168 OxdC (Yvrk), which shared a high amino acid similarity but showed limited demulsification ability. Based on a comparison of the structural characteristics, the hydrophobic amino acids on the surface of OxdC were identified as a key factor driving the favorable demulsification activity of XH1. The metabolic pathways of XH1 used liquid paraffin and glucose as substrates, illustrating that hydrocarbons are necessary for biodemulsifier secretion. The present study provides new insight into the application of OxdC as an additional genetic resource in biodemulsification.
研究领域:
微生物学
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环境生物技术
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生物化学
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基因组学
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CHEMICAL ENGINEERING JOURNAL
·
2020
·
IF 10.652
Yunxiu Liu,Yu Tian,Qiuyang Han,Jie Yin,Junchuan Zhang,Yue Yu,Weizhong Yang,Yi Deng
Because of the explosive generation of drug-fast bacteria, antibiotics-free strategy is much-needed for elimination of bacterial-associated contagion. Herein, we develop a near-infrared (NIR) activated heterostructure catalyst (MXene/CoNWs) consisted of two-dimensional (2D) MXene and one-dimensional (1D) cobalt nanowires (CoNWs) in acting against bacteria for drug-free therapy. The intercalated CoNWs can rapidly trap the photogenerated electrons from Ti 3 C 2 MXene nanosheets, thus restrains the recombination of hot electron-hole and facilitates transfer of carriers upon 808 nm NIR illumination, thereby increasing the yields of germicidal reactive oxygen species (ROS). Additionally, the photothermal effect of the 2D/1D heterostructure is significantly strengthened due to the synergy of plasmonic CoNWs and MXene semiconductor. The heterostructure coatings on an orthopedic implant can achieve a high antibacterial efficacy of over 90% against Gram-positive/Gram-negative bacteria within 20 min, resulting from the synergistic actions of NIR-triggered ROS and hyperthermia. Besides, in vitro findings indicate that MXene/CoNWs heterojunctions show acceptable cytocompatibility. Accordingly, the two merits make the novel MXene/CoNWs heterojunctions become a promising drug-free antibacterial therapy for cleaning the pathogenic infection and avoiding antibiotic resistance in practice.
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ENZYME AND MICROBIAL TECHNOLOGY
·
2021
·
IF 3.493
Li-Bin Guo,Chen-Yuan Zhu,Yi-Bei Wu,Xiao-Man Fan,Ye-Wang Zhang
Chondroitin AC lyase can efficiently hydrolyze chondroitin sulfate (CS) to low molecule weight chondroitin sulfate, which has been widely used in clinical therapy, including anti-tumor, anti-oxidation, hypolipidemic, and anti-inflammatory. In this work, a novel chondroitin AC lyase from Pedobacter xixiisoli ( Px chonAC) was cloned and overexpressed in Escherichia coli BL21 (DE3). The characterization of Px chonAC showed that it has specific activities on chondroitin sulfate A, Chondroitin sulfate C and hyaluronic acid with 428.77, 270.57, and 136.06 U mg −1 , respectively. The K m and V max of Px chonAC were 0.61 mg mL -1 and 670.18 U mg -1 using chondroitin sulfate A as the substrate. The enzyme had a half-life of roughly 660 min at 37 °C in the presence of Ca 2+ and remained a residual activity of 54 % after incubated at 4 °C for 25 days. Molecular docking revealed that Asn123, His223, Tyr232, Arg286, Arg290, Asn372, and Glu374 were mainly involved in the substrate binding. The enzymatic hydrolysis product was analyzed by gel permeation chromatography, demonstrating Px chonAC could hydrolyze CS efficiently.
研究领域:
生物化学
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酶学
·
药理学
·
微生物学
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PubMed
高影响因子
危险品化学品经营许可证（带存储）