Molecular Catalysis
·
2025
·
IF 4.9
Kangping Huang,Wei Song,Wanqing Wei,Guipeng Hu,Xiaomin Li,Cong Gao,Jing Wu
Salvianic acid A (SAA), a bioactive compound from Salvia miltiorrhiza , has attracted significant attention for its therapeutic properties, including antitumor and antioxidant activities. However, its large-scale application is limited by the conventional production methods, which rely primarily on physical extraction and chemical synthesis. In this study, we constructed a de novo biosynthetic pathway for SAA by integrating two key modules: the HPPA hydroxylation module HpaBC (4-hydroxyphenylacetate 3-monooxygenase from Escherichia coli ) and the DHPPA reductase module Rg MDH (D-mandelic acid dehydrogenase from Rhodotorula graminis ) into a high-yield tyrosine-producing strain, Tyr0 , generating the engineered strain Dps01. Metabolic analysis identified the low catalytic efficiency of Rg MDH as the critical bottleneck. To address this, structure-guided protein engineering of Rg MDH was performed. Molecular dynamics (MD) simulations revealed that the optimal mutant Rg MDH M4 (T312A/F316S/T255I/A59H) exhibited a 10.3-fold increase in catalytic conformation occupancy compared to the wild-type (4.74% vs 0.46%), indicating enhanced substrate channel dynamics. This mutant exhibited a 295% increase in activity toward DHPPA, with enhanced substrate affinity and a 5.76-fold improvement in catalytic efficiency ( k cat / K m ). When integrated into strain Dps02, the engineered system achieved a maximum SAA production of 6.82 g/L within 34 hours, with a productivity of 0.21 g/L·h⁻¹ in a 5 L fed-batch fermentation. These results represent a significant advancement in the biosynthesis of SAA.
研究领域:
生物化学
·
合成生物学
·
药物开发
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DOI
高影响因子
Theranostics
·
2025
·Vol. 15
·期 1
·68
·
IF 13.3
Mingbo Wu,Zhanlin Zhang,Dong Li,Xiaomiao Ruan,Jingwen Yang,Siyi Chen,Xin Li,Wenwu Ling
Rationale: Ferroptosis and sonodynamic therapy (SDT) are both promising therapeutic modalities, but their clinical application remains challenging due to the hypoxic tumor microenvironment and limited supply of polyunsaturated fatty acids. Developing an agent with oxygen-enhanced SDT and increased ferroptosis sensitivity is crucial for advancing tumor therapy. Methods: In this study, catalase (Cat) and Acyl-CoA synthetase long-chain family member 4 (ACSL4) highly expressed 4T1 cells were constructed via lentivirus transfection. Cat and ACSL4 enriched exosomes (EXO@CA) were then extracted and loaded with the sonosensitizer tetrakis (4-carboxyphenyl) porphyrin (TCPP) through electroporation to create engineered exosomes (EXO@CAT). We evaluated the ability of EXO@CAT to generate oxygen in a hydrogen peroxide environment and investigated its effect on motion profiles and permeability of EXO@CAT. The in vitro antitumor activity was assessed via cytotoxicity, ROS levels, live/dead staining, and apoptosis, with ferroptosis biomarkers confirming ferroptosis activation. We also evaluated the in vivo anticancer efficacy of EXO@CAT by tumor growth analysis and histological and immunohistochemical staining in mouse models bearing breast tumor. Results: EXO@CAT harnesses ultrasound stimulation to facilitate oxygen-enriched SDT, demonstrating significant capacity for singlet oxygen ( 1 O 2 ) generating, which promotes the accumulation of lipid peroxidation (LPO), ultimately leading to the induction of ferroptosis. Concurrently, ACSL4 released from EXO@CAT also increases LPO accumulation by modifying cellular lipid composition, thereby enhancing cellular sensitivity to ferroptosis. Moreover, both in vitro and in vivo experiments demonstrate that the homologous targeting ability of EXO@CAT enables its efficient accumulation in tumor tissues, and the oxygen generation catalyzed by Cat not only alleviates tumor hypoxia but also facilitates the penetration of EXO@CAT into deeper layers of tumor tissue. Conclusions: EXO@CAT combines endogenous proteins, which are prone to inactivation, with an exogenous sonosensitizer, allowing synergistic anticancer treatment of both ferroptosis and SDT with improved efficacy.
CAS / MDL:
研究领域:
肿瘤学
·
生物医学工程
·
细胞生物学
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产品关联
DOI
PubMed
高影响因子
PROCESS BIOCHEMISTRY
·
2018
·
IF 2.616
Liqing Zhao,Yingmiao Xie,Liuyan Chen,Xuefeng Xu,Cindy Xinxin Zhao,Feng Cheng
Biotransformation of isoeugenol to vanillin is an attractive approach in industry for vanillin synthesis. However, the biotransformation efficiency is hampered by the low activity of isoeugenol monooxygenase (IEM) and the product inhibition towards IEM. Therefore, it is worth discovering a new IEM and improving its enzymatic properties by protein engineering. Herein, a new IEM720 gene was firstly cloned from microbial metagenome and successfully expressed in E. coli . Secondly, five potentially beneficial amino acid positions were selected by a “three-criteria” in silico system and visual inspection was performed for grouping them into two sets (regions A and B). The variant IEM720-F281Q displayed highest isoeugenol conversion and k cat value after screening the site-saturation libraries of these five positions. Furthermore, the structure-functional relationships of the beneficial substitution on IEM activity were investigated. The final vanillin concentration reached ∼4.5 g/L (∼75% conversion) by combining sol-gel chitosan membrane (for relieve of product inhibition) and E. coli cells harboring IEM720-F281Q.
研究领域:
生物化学
·
分子生物学
·
工业微生物学
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DOI
高影响因子
JOURNAL OF HAZARDOUS MATERIALS
·
2019
·
IF 7.65
Peng Liao,Binrui Li,Lin Xie,Xiaoao Bai,Han Qiao,Qianqian Li,Biwei Yang,Chongxuan Liu
Engineered nanoparticles-mediated contaminant transport has been recognized as a significant process governing the mobility of metals and radionuclides in groundwater. Engineered silicate nanoparticles (ESNPs) are attractive materials for the sequestration or extraction of Cr(VI) and other metals and radionuclides from groundwater. While great efforts have been devoted toward the application of these materials for Cr(VI) sequestration, the underlying interface adsorption mechanism is not thoroughly elucidated. This study investigates the immobilization mechanisms of Cr(VI) on a representative ESNPs, NH 2 -MCM-41, over a range of water chemistry conditions. By combining batch adsorption experiments with an array of complementary characterizations, we provided spectroscopic and microscopic evidence that the electrostatic interactions between the positively charged NH 2 -MCM-41 surface derived from amino functionality and the negatively charged Cr(VI) species was the dominant mechanism responsible for Cr(VI) immobilization. In addition, the weak hydrogen bonding interactions may also contribute to adsorption to a degree. Furthermore, thermodynamic studies suggested a favorable, spontaneous, and exothermic adsorption process. Site energy analysis illustrated that the distribution of energy binding sites on NH 2 -MCM-41 is Cr(VI) loading dependent. The new insights provided here can advance understanding of the transport of Cr(VI) associated NH 2 -MCM-41 that benefits the application of ESNPs-based technologies for metals immobilization in groundwater.
研究领域:
环境科学
·
纳米技术
·
污染治理
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DOI
PubMed
高影响因子
PROCESS BIOCHEMISTRY
·
2025
·
IF 4
Jing-Jing Liu,Bin-Bin Jiang,Ya-Han Zhu,Shu-Qin Gao,Ying-Wu Lin
Azo dyes, widely used in global textiles, persist in wastewater causing significant ecological and health risks, necessitating efficient degradation to protect ecosystems and public health. The development of artificial metalloenzymes has provided advanced biotechnological solutions to address this challenge. This study presents a highly efficient artificial metalloenzyme, F43Y/H64D myoglobin (F43Y/H64D Mb), engineered from sperm whale myoglobin with a modified heme active site by replacement of Phe43 with Tyr and His64 with Asp, respectively, for azo dye degradation. The engineered enzyme achieved degradation efficiencies of approximately 93%, 93%, and 90% for Acid Orange 7, Methyl Orange, and Acid Blue 92, respectively, within 10 seconds. Its overall catalytic efficiency was found to be approximately 3.6 times higher than that of horseradish peroxidase, a widely used natural enzyme. HPLC and ESI-MS analyses proposed degradation pathways, and bacterial toxicity tests confirmed reduced toxicity post-degradation. Furthermore, F43Y/H64D Mb exhibited excellent performance in natural aquatic environments, surpassing many recently reported results in efficiency, reaction speed, and operational simplicity. These findings highlight the feasibility of F43Y/H64D Mb for efficient azo dye decolorization and degradation in wastewater, offering a novel enzymatic tool for sustainable environmental remediation.
研究领域:
环境科学
·
生物化学
·
酶工程
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产品关联
DOI
高影响因子
ADVANCED MATERIALS
·
2024
·Vol. 36
·期 30
·2404629
·
IF 27.4
Jing Liang,Xin Zhang,Haidong Li,Chuanxi Wen,Longlong Tian,Ximeng Chen,Zhan Li
The special properties of rare earth elements (REE) have effectively broadened their application fields. How to accurately recognize and efficiently separate target rare earth ions with similar radii and chemical properties remains a formidable challenge. Here, precise two-dimensional (2D) heterogeneous channels are constructed using engineered E. coli membranes between graphene oxide (GO) layers. The difference in binding ability and corresponding conformational change between Lanmodulin (LanM) and rare earth ions in the heterogeneous channel allows for precisely recognizing and sieving of scandium ions (Sc 3+ ). The engineered E. coli membranes not only can protect the integrity of structure and functionality of LanM, the rich lipids and sugars, but also help the Escherichia coli ( E. coli ) membranes closely tile on the GO nanosheets through interaction, preventing swelling and controlling interlayer spacing accurately down to the sub-nanometer. Apparently, the 2D heterogeneous channels showcase excellent selectivity for trivalent ions (SF Fe /Sc ≈3), especially for Sc 3+ ions in REE with high selectivity (SF Ce/Sc ≈167, SF La/Sc ≈103). The long-term stability and tensile strain tests verify the membrane's outstanding stability. Thus, this simple, efficient, and cost-effective work provides a suggestion for constructing 2D interlayer heterogeneous channels for precise sieving, and this valuable strategy is proposed for the efficient extraction of Sc.
研究领域:
材料科学
·
生物化学
·
分离技术
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产品关联
DOI
PubMed
高影响因子
ACS Central Science
·
2024
·
IF 12.7
Yujie Wang,Zhengao Di,Minglang Qin,Shenming Qu,Wenbo Zhong,Lingfeng Yuan,Jing Zhang,Julian M. Hibberd,Ziyi Yu
In this study, an innovative approach is presented in the field of engineered plant living materials (EPLMs), leveraging a sophisticated interplay between synthetic biology and engineering. We detail a 3D bioprinting technique for the precise spatial patterning and genetic transformation of the tobacco BY-2 cell line within custom-engineered granular hydrogel scaffolds. Our methodology involves the integration of biocompatible hydrogel microparticles (HMPs) primed for 3D bioprinting with Agrobacterium tumefaciens capable of plant cell transfection, serving as the backbone for the simultaneous growth and transformation of tobacco BY-2 cells. This system facilitates the concurrent growth and genetic modification of tobacco BY-2 cells within our specially designed scaffolds. These scaffolds enable the cells to develop into predefined patterns while remaining conducive to the uptake of exogenous DNA. We showcase the versatility of this technology by fabricating EPLMs with unique structural and functional properties, exemplified by EPLMs exhibiting distinct pigmentation patterns. These patterns are achieved through the integration of the betalain biosynthetic pathway into tobacco BY-2 cells. Overall, our study represents a groundbreaking shift in the convergence of materials science and plant synthetic biology, offering promising avenues for the evolution of sustainable, adaptive, and responsive living material systems.
研究领域:
植物学
·
合成生物学
·
材料科学
·
生物工程
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产品关联
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2024
·
IF 13.3
Jing Liu,Yize Li,Dongxu Wang,Xinyu Liu,Dawei Sun,Yixin Wang,Yingying Zhang,Hanjie Wang
Cyanobacteria, due to its excellent oxygen production capability and desirable biocompatibility, have promising application prospects in oxygen-associated therapy, such as photodynamic therapy (PDT). Cyanobacteria-based PDT has emerged as an attractive microbial therapy. However, as the limited loading capacity of photosensitizer on cyanobacteria, it’s difficult to maintain a long and stable efficiency for cyanobacteria-based PDT. To break through this limitation, we designed and developed an engineered cyanobacteria (ECyano) using synthetic biology techniques. ECyano not only relieved the hypoxic tumour microenvironment, but also synthesized and accumulated 5-aminolevulinic acid (5-ALA) in tumour, generating reactive oxygen species (ROS) for killing tumours under laser irradiation. Then we attached manganese dioxide (MnO 2 ) nanoparticles to ECyano, thus creating a novel synergistic functional nano-biosystem, named as ECyano@MnO 2 . The combination of ECyano and MnO 2 was tuned to achieve PDT-immunotherapy. Upon the irradiation of 640 nm laser, ECyano@MnO 2 could firstly achieve self-supplying photosensitizer and oxygen, achieving the therapeutic effect. And then MnO 2 could act as an excellent adjuvant which could be degraded into Mn 2+ in tumour microenvironment and dendritic cells (DCs) were stimulated, eliciting potent anti-tumour immunity. The in vivo results showed partial tumour suppression of 79.34% treated with ECyano@MnO 2 . These results showed that engineered cyanobacteria-based nano-biosystem provided a promising strategy against tumour and may open the door to a promising microbial PDT.
研究领域:
生物医学工程
·
合成生物学
·
肿瘤学
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DOI
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2025
·
IF 8.5
Qianqian Jiang,Qi Chu,Qi Zhang,Chaoqiang Wu,Tongqing Tang,Xin Wang,Feifei Chen,Alei Zhang,Kequan Chen
Adol condensation mediated by L-threonine aldolase (LTA) is a promising route to synthesize β-hydroxy-α-amino acids, such as the anti-Parkinson's disease drug L- threo -3,4-dihydroxyphenylserine (L- threo -DOPS; droxidopa). However, the biocatalytic approach generally suffered from poor stereoselectivity at the C β chiral center. Herein, several LTAs from diverse microorganisms were examined for the stereoselective synthesis of L- threo -DOPS, and an LTA from Firmicutes bacterium ( Fb LTA) was identified with relatively high stereoselectivity. A semi-rational design was then employed to further enhance the stereoselectivity, resulting in the identification of beneficial mutations and the generation of beneficial mutants with markedly elevated stereoselectivity (up to 80 % de) over the wild-type (23 % de). Molecular dynamics (MD) simulation revealed that the significant improvement of stereoselectivity could be attributed to the structural reconstitution of substrate tunnels. Finally, the best mutant was applied to a scaled-up reaction, affording 1.3 g/L yield of L- threo -DOPS with 80 % de. This study provides new insights into the stereoselectivity control of LTAs and potential engineered LTAs for the manufacture of L- threo -DOPS.
研究领域:
生物化学
·
合成生物学
·
药物开发
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产品关联
DOI
PubMed
高影响因子
Scientific Reports
·
2016
·Vol. 6
·期 1
·1-11
·
IF 5.228
Liu Hongmei,Chang Hong,Lv Jia,Jiang Cong,Li Zhenxi,Wang Fei,Wang Hui,Wang Mingming,Liu Chongyi,Wang Xinyu,Shao Naimin,He Bingwei,Shen Wanwan,Zhang Qiang,Cheng Yiyun
Polymers are widely used as non-viral carriers for siRNA delivery, but concern has also arisen in their limited efficacy and inherent toxicity. Whilst many of previous efforts have been documented towards improving the performance of polymers via chemical modifications, the structure-activity relationships (SAR) of these ligand-modified polymers are not well understood. To address this issue, we systemically prepared a library of surface-engineered dendrimers (>300) as the screening pool to discover efficient siRNA carriers. The modified ligands include alkyls and fluoroalkyls, amino acids, benzene derivatives and heterocyclic compounds. Gene silencing results showed that the lead material shows excellent efficacy even in hard-to-transfect cells such as mesenchymal stem cells. The SAR studies revealed that ligands containing appropriate hydrophobicity, or ligands with both hydrophobic and functional atoms/groups are essential for polymers to achive efficient knockdown efficacy. A second-generation library designed based on the above principles further confirms the proposed design criteria. The results enable the future rational design of potent siRNA carriers.
研究领域:
药学
·
分子生物学
·
纳米技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF ALLOYS AND COMPOUNDS
·
2025
·
IF 6.3
Huangtong Chen,Yifei Yang,Xiaofei Li,Huimei Duan
Defect engineering in oxide supports has emerged as a promising approach to enhance the surface chemical properties of active metals for methane catalytic oxidation. Herein, the N-doped Pd/Co 3 O 4 (Pd/N-Co 3 O 4 ) catalysts were successfully synthesized using ZIF-67 as a self-sacrificial template via an H 2 O-soaking treatment. Compared to conventional Pd/Co 3 O 4 catalysts (T 50 : 294 °C), the Pd/N-Co 3 O 4 catalysts exhibited superior catalytic activity (T 50 : 278 °C) and exceptional water resistance, with only a 6% activity loss at 350 °C in the presence of 5 vol% H 2 O, as opposed to a 56% loss for Pd/Co 3 O 4 catalysts. Detailed experimental characterizations revealed that N-doping significantly increased oxygen vacancy concentrations and surface lattice oxygen mobility while stabilizing Pd/PdO x active sites through the electronic metal-support interaction (EMSI) between Pd and N-Co 3 O 4 . In the Pd/N-Co 3 O 4 catalysts, the enhanced oxygen vacancies rendered the N-Co 3 O 4 support electron-rich, which in turn diminishes the electron transfer from Pd to N-Co 3 O 4 supports. This modulation effectively inhibited the excessive oxidation of Pd/PdO x active species into bulk PdO, thereby enhancing catalytic performance and water tolerance in methane oxidation. These findings provide valuable insights into the role of N-doped defect engineering in facilitating efficient methane catalytic combustion, highlighting the significance of EMSI and oxygen vacancies in optimizing catalyst performance.
研究领域:
催化
·
材料科学
·
环境工程
·
化学工程
·
表面科学
✓ 已验证
产品关联
DOI
高影响因子
Materials Today Bio
·
2024
·
IF 8.7
Qing Xia,Feila Liu,Yue Zhou,Guanyuan Yang,Fangzhou Li,Tingting Liang,Jun Liu,Wanling Li,Yaqing Huang,Chuhong Zhu
The accumulation of foam cells in the subendothelial space of the vascular wall to form plaques is the real cause of atherosclerotic lesions. Conventional interventions, such as statins and anti-cytokine or anti-inflammatory therapies, suffer problems in terms of their short therapeutic outcomes and potential disruption of the immune system. The development of more efficient therapeutics to restrict the initial progression of plaques appears to be crucial for treating and preventing atherosclerosis. Decreasing foam cell formation by reversing the excessive phagocytosis of modified low-density lipoprotein (LDL) in macrophages is highly desirable. Here, we developed a strategy based on engineered monocytes to dynamically regulate lipid uptake by macrophages inspired by a CD47–SIRPα signaling-induced defect in the phagocytosis of lesional macrophages at the advanced stage of AS. Briefly, a complex called CD47p-GQDs-miR223, which is designed to interact with SIRPα, was synthesized to remodel monocytes by decreasing the uptake of oxidized LDL through the activation of CD47-SIRPα signaling. After injection, these monocytes compete for recruitment to atherosclerotic plaques, release gene drugs and mediate anti-inflammatory phenotypic remodeling of the aboriginal macrophages, effectively inhibiting the development of foam cells. Our strategy provides a new therapeutic for preventing the progression of atherosclerosis.
研究领域:
心血管疾病
·
分子生物学
·
治疗学
·
免疫学
✓ 已验证
DOI
高影响因子
危险品化学品经营许可证（带存储）