Scientific Reports
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2017
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IF 4.259
Kunpeng Li,Rongzhen Zhang,Yan Xu,Zhimeng Wu,Jing Li,Xiaotian Zhou,Jiawei Jiang,Haiyan Liu,Rong Xiao
(S)-carbonyl reductase II (SCRII) from Candida parapsilosis is a short-chain alcohol dehydrogenase/reductase. It catalyses the conversion of 2-hydroxyacetophenone to (S)-1-phenyl-1,2-ethanediol with low efficiency. Sortase was reported as a molecular "stapler" for site-specific protein conjugation to strengthen or add protein functionality. Here, we describe Staphylococcus aureus sortase A-mediated crosslinking of SCRII to produce stable catalysts for efficient biotransformation. Via a native N-terminal glycine and an added GGGGSLPETGG peptide at C-terminus of SCRII, SCRII subunits were conjugated by sortase A to form crosslinked SCRII, mainly dimers and trimers. The crosslinked SCRII showed over 6-fold and 4-fold increases, respectively, in activity and k cat /K m values toward 2-hydroxyacetophenone compared with wild-type SCRII. Moreover, crosslinked SCRII was much more thermostable with its denaturation temperature (T m ) increased to 60 °C. Biotransformation result showed that crosslinked SCRII gave a product optical purity of 100% and a yield of >99.9% within 3 h, a 16-fold decrease in transformation duration with respect to Escherichia coli/pET-SCRII. Sortase A-catalysed ligation also obviously improved T m s and product yields of eight other short-chain alcohol dehydrogenases/reductases. This work demonstrates a generic technology to improve enzyme function and thermostability through sortase A-mediated crosslinking of oxidoreductases.
研究领域:
生物化学
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酶工程
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蛋白质工程
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DOI
PubMed
高影响因子
Carbon Letters
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2023
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IF 4.5
Li Guochun,Li Jiaxin,Yang Ziyu
This study reports the synthesis of a novel graphene/chitosan/β-cyclodextrin composite material (GO/CS/β-CD) via a one-step chemical reduction method, which combines the advantages of graphene, chitosan, and β-cyclodextrin. The morphology and structure of the composite were characterized using various techniques, such as scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and Raman spectroscopy. Subsequently, sortase A (SA) was immobilized onto the GO/CS/β-CD for the detection of Staphylococcus aureus . The sensor exhibited a good linear relationship within the concentration range of 30–300 CFU/mL, with a detection limit of 12 CFU/mL. The GO/CS/β-CD composite material showed enhanced properties due to the synergistic effect of graphene, chitosan, and β-cyclodextrin. The immobilization of sortase A onto the composite material improved the sensitivity and selectivity of the sensor for the detection of S. aureus . This study presents a novel graphene/chitosan/β-cyclodextrin composite material with immobilized sortase A, demonstrating enhanced sensitivity and selectivity for the detection of Staphylococcus aureus, which has potential for the development of high-performance sensors in various fields.
研究领域:
生物传感
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微生物学
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材料科学
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DOI
高影响因子
RSC Advances
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2025
·Vol. 15
·期 41
·33966-33975
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IF 4.6
Yin-Yu Qi,Lu-Yang Han,Long-Xu Han,Wen-Han Bu,Yang Xu,Jian-Jun Chu
A sortase A inhibitor (SrtA I)-loaded bone cement (SLBC) containing 1-(3,4-dichlorophenyl)-3-(dimethylamino)propyl-1-one hydrochloride (AEEK1) was reported. Compared with gentamicin-loaded bone cement (GS cement), which is a widely used antibiotic-loaded bone cement in cemented total joint replacement (TJR), SLBC demonstrated a significantly prolonged effective antibacterial duration against both Staphylococcus aureus (S. aureus) and methicillin-resistant S. aureus (MRSA). The quasi-static compressive strength, maximum polymerization temperature, and setting time of SLBC were within the limits stated in the ISO 5833 standard and comparable to the corresponding values of GS cement. In vitro and in vivo experimental results confirmed the biocompatibility of SLBC and showed its easy injectability into porcine T12 vertebrae. Taken together, these findings suggest that SLBC may have potential for managing/treating prosthetic joint infections, which is a common complication in cemented TJR.
研究领域:
生物材料
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骨科手术
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感染性疾病
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药物制剂
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生物医学工程
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DOI
PubMed
高影响因子
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
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2024
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IF 3.9
Yujia Chen,Wei Li,Li Wang,Bingmei Wang,Jian Suo
In confronting the significant challenge posed by multidrug-resistant (MDR) pathogens, particularly methicillin-resistant Staphylococcus aureus (MRSA), the development of innovative anti-infective strategies is essential. Our research focuses on sortase A (SrtA), a vital enzyme for anchoring surface proteins in S. aureus. We discovered that plantamajoside (PMS), a phenylpropanoid glycoside extracted from Plantago asiatica L. (Plantaginaceae), acts as an effective and reversible inhibitor of SrtA, with a notable IC50 value of 22.93 µg/mL. This breakthrough provides a novel approach to combat both resistance and virulence in MRSA. PMS significantly inhibits S. aureus adhesion to fibrinogen, reducing biofilm formation and hindering the anchoring of staphylococcal protein A to the cell wall. Live-dead cell assays demonstrated increased survival rates in PMS-treated MRSA-infected A549 cells. Fluorescence quenching experiments revealed a robust interaction between PMS and SrtA, with mechanistic analyses pinpointing the critical R197 amino acid residue as the target site. In vivo, PMS was highly effective in a Galleria mellonella infection model, reducing mortality rates in MRSA-infected larvae. Additionally, PMS demonstrated therapeutic efficacy in a mouse pneumonia model, improved survival rates, reduced the bacterial load in pulmonary tissues, and mitigated lung damage. These results validate PMS as a promising compound to mitigate MRSA virulence and thwart resistance by targeting SrtA. This study highlights PMS as a leading candidate for controlling MRSA infections, showing the potential of targeting specific bacterial mechanisms in the fight against MDR infections.
研究领域:
微生物学
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药理学
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传染病学
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分子生物学
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DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2024
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IF 5.7
Zhuohui Zhang,Yan Wang,Qianqian Xu,Yimiao Liu,Kaiyu Wang,Pishun Li,Xiaofeng Zheng
Methicillin-resistant Staphylococcus aureus (MRSA), one of the most important zoonotic bacterial pathogens, necessitates innovative antibacterial strategies. Rosmarinic acid (RA) possesses various biological functions, including antibacterial and anti-inflammatory effects. This study aimed to explore the synergistic effects and mechanism of RA in conjunction with ceftiofur (CF) against MRSA. Results showed that the combination of RA and CF demonstrated synergistic anti-MRSA activity, conferring protective effects in an MRSA bacteremia mouse model. Mechanistic investigations further established that RA interacts with sortase A (SrtA) through hydrogen bonds, electrostatic, and van der Waals forces at critical sites, leading to the reversible inhibition of SrtA activity. Consequently, this inhibition impeded the ability of MRSA to invade cells and form biofilms. Conclusively, RA directly inhibits SrtA activity, thus enhancing the synergistic anti-MRSA effect of CF. These findings provide a theoretical foundation for the use of RA in combination with CF as a therapeutic strategy for MRSA infections.
研究领域:
微生物学
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药理学
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感染病学
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产品关联
DOI
PubMed
高影响因子
BIOCONJUGATE CHEMISTRY
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2026
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IF 4.5
Liqiang Wei,Hongfei Wang,Chunyue Du,Antony Kam,Shining Loo
We present a one-pot approach for orthogonal dual functionalization of mi3 self-assembling protein nanoparticles by combining SpyCatcher003/SpyTag003 isopeptide bond formation with Sortase A-mediated transpeptidation. The nanoparticles were engineered with N-terminal SpyCatcher003 domains and C-terminal LPETGGH motifs, enabling site-specific conjugation of distinct cargo molecules. To overcome expression limitations, an N-terminal vesicle nucleating peptide (VNP6) was incorporated, which enhanced protein yields while maintaining nanocage integrity. Critical to this strategy, the calcium-independent Sortase A variant (SRT7+) was employed to avoid calcium-induced nanoparticle aggregation, enabling orthogonal ligation under physiological conditions. The platform enabled simultaneous conjugation of HER2-targeting affibodies and fluorescent probes in a single reaction, and these dual-functionalized self-assembling protein nanoparticles exhibited specific cellular uptake in HER2-positive cancer cells. This integrated one-pot approach proved scalable and modular for engineering protein-based nanomaterials for targeted therapy and diagnostics.
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DOI
PubMed
高影响因子
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