Enhanced anticoagulant activity of hirudin-i analogue co-expressed with arylsulfotransferase in periplasm of E. coli BL21(DE3)

Rai, Kamal; Chu, Xiaohui; Bao, Zixian; Liang, Yunlong; Wang, Xingang; Yang, Junqing; Xian, Ming; Sun, Yue; Nian, Rui

doi:10.1016/j.jbiotec.2020.08.003

Cited by 9 publications

(9 citation statements)

References 36 publications

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“…Co-expressed rH-I analogue with sulfate donor substrate (p-nitrophenyl sulfate potassium) showed anticoagulant (rabbit and goat serum) activity twice more than rH-I analogue expressed without ASST. In addition, purified rH-I analogue showed above 4.5 times higher anticoagulant activity compared to therapeutic anti-thrombotic heparin (Rai et al, 2020). These research progress of hirudin in structure modification may greatly promote its clinical application and overcome its original defects.…”

Section: Increasing the Antithrombotic Efficacymentioning

confidence: 95%

Pharmacological Activities and Mechanisms of Hirudin and Its Derivatives - A Review

et al. 2021

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Hirudin, an acidic polypeptide secreted by the salivary glands of Hirudo medicinalis (also known as “Shuizhi” in traditional Chinese medicine), is the strongest natural specific inhibitor of thrombin found so far. Hirudin has been demonstrated to possess potent anti-thrombotic effect in previous studies. Recently, increasing researches have focused on the anti-thrombotic activity of the derivatives of hirudin, mainly because these derivatives have stronger antithrombotic activity and lower bleeding risk. Additionally, various bioactivities of hirudin have been reported as well, including wound repair effect, anti-fibrosis effect, effect on diabetic complications, anti-tumor effect, anti-hyperuricemia effect, effect on cerebral hemorrhage, and others. Therefore, by collecting and summarizing publications from the recent two decades, the pharmacological activities, pharmacokinetics, novel preparations and derivatives, as well as toxicity of hirudin were systematically reviewed in this paper. In addition, the clinical application, the underlying mechanisms of pharmacological effects, the dose-effect relationship, and the development potential in new drug research of hirudin were discussed on the purpose of providing new ideas for application of hirudin in treating related diseases.

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Section: Increasing the Antithrombotic Efficacymentioning

confidence: 95%

Pharmacological Activities and Mechanisms of Hirudin and Its Derivatives - A Review

et al. 2021

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“…Hiru is a direct inhibitor of thrombin, and its role in human diseases has been highlighted by emerging reports [ 13 ]. For instance, Yu et al have proposed that Hiru mitigates Ang II-stimulated myocardial fibroblast fibrosis through inactivating the ERK1/2 pathway.…”

Section: Discussionmentioning

confidence: 99%

“…Hirudin (Hiru), a peptide naturally occurring in leeches and their salivary glands, possesses anticoagulative property [ 13 ]. Also, more attention has been paid on the anti-inflammatory effects of Hiru.…”

Section: Introductionmentioning

confidence: 99%

Hirudin Regulates Vascular Function in Chronic Renal Failure through Modulating Macrophage Polarization

Chen¹,

Ding²,

Yang³

2022

BioMed Research International

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Excessive inflammation is responsible for arteriovenous fistula (AVF) failure, which determines the therapeutic effect of chronic renal failure (CRF). Macrophage polarization is of great significance in the inflammatory response. Hirudin (Hiru) has been reported to possess a definite anti-inflammatory effect. This study is to uncover the impacts of Hiru on classically (M1)/alternatively (M2) macrophage polarization in the CRF rat model and rat vascular smooth muscle cells (VSMCs). After the CRF rat model was administrated with different concentrations of Hiru, blood urea nitrogen (BUN) and serum creatinine (Scr) levels were tested. H&E staining was to detect vascular injury, and IHC assay was to analyze inducible nitric oxide synthase (iNOS) and arginase-1 (Arg-1) expressions in vascular tissues. Levels of inflammatory factors were examined by ELISA. Besides, western blot was to estimate the levels of marker proteins related to macrophage, proliferation, and apoptosis. CCK-8 was to measure cell viability. We discovered that Hiru alleviated renal function injury and vascular injury, exacerbated VSMC hyperplasia, and stimulated the differentiation and activation of M1 macrophage towards M2 macrophage in vivo. Moreover, after treatment with lipopolysaccharide (LPS)/IFN-gamma (IFN-γ), the increased M1/M2 ratio and enhanced levels of inflammatory factors were observed. Furthermore, Hiru boosted the proliferation and ameliorated the inflammatory response and apoptosis of rat VSMCs during the process of coincubation of M1-conditioned medium (CM). Collectively, Hiru played a protective role against vascular injury in CRF directly or through its influence on M1 macrophage polarization and inflammation.

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“…Cell fractionation was performed according to the method described by Quan [ 32 , 33 ]. Cells were harvested from 10 mL culture broth (OD600 ≈ 1.6) by centrifugation (5000 g, 10 min, 4 °C) and treated with one-third volume of Tris-EDTA-NaCl solution (TEN) containing 50 mM Tris-HCl, 5 mM EDTA, and 50 mM NaCl (pH 8.0).…”

Section: Methodsmentioning

confidence: 99%

Whole-Cell Display of Phosphotransferase in Escherichia coli for High-Efficiency Extracellular ATP Production

Zhao¹,

Yang²,

Xie³

et al. 2022

Biomolecules

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Adenosine triphosphate (ATP), as a universal energy currency, takes a central role in many biochemical reactions with potential for the synthesis of numerous high-value products. However, the high cost of ATP limits industrial ATP-dependent enzyme-catalyzed reactions. Here, we investigated the effect of cell-surface display of phosphotransferase on ATP regeneration in recombinant Escherichia coli. By N-terminal fusion of the super-folder green fluorescent protein (sfGFP), we successfully displayed the phosphotransferase of Pseudomonas brassicacearum (PAP-Pb) on the surface of E. coli cells. The catalytic activity of sfGFP-PAP-Pb intact cells was 2.12 and 1.47 times higher than that of PAP-Pb intact cells, when the substrate was AMP and ADP, respectively. The conversion of ATP from AMP or ADP were up to 97.5% and 80.1% respectively when catalyzed by the surface-displayed enzyme at 37 °C for only 20 min. The whole-cell catalyst was very stable, and the enzyme activity of the whole cell was maintained above 40% after 40 rounds of recovery. Under this condition, 49.01 mg/mL (96.66 mM) ATP was accumulated for multi-rounds reaction. This ATP regeneration system has the characteristics of low cost, long lifetime, flexible compatibility, and great robustness.

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Enhanced anticoagulant activity of hirudin-i analogue co-expressed with arylsulfotransferase in periplasm of E. coli BL21(DE3)

Cited by 9 publications

References 36 publications

Pharmacological Activities and Mechanisms of Hirudin and Its Derivatives - A Review

Pharmacological Activities and Mechanisms of Hirudin and Its Derivatives - A Review

Hirudin Regulates Vascular Function in Chronic Renal Failure through Modulating Macrophage Polarization

Whole-Cell Display of Phosphotransferase in Escherichia coli for High-Efficiency Extracellular ATP Production

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