In the present investigation, a lipid hydrolyzing gene RPK01 was cloned from metagenome source of hot spring. Expression and purification of recombinant protein revealed single purified protein band of ~24 KDa on 12% SDS-PAGE, and is well corroborated with the deduced molecular weight of protein as calculated from its amino acid sequence. The purified protein displayed high activity towards short chain fatty acids and was found to be completely stable at 30°C till 3h, it further retained ~40% activity at 50°C and 60°C temperature till 3h. Additionally, the pH stability assay showed its functionality in broad range of pH, with maximum stability observed at pH 2.0, it decreases from pH 4.0 to pH 12.0, and nearly showed 40% activity in these pH values. Both circular dichroism and intrinsic Trp fluorescence studies revealed conformational stability of protein structure at wide range of temperature and pH. Enzyme activity enhances in presence of non-ionic surfactants like Tween 20 and TritonX-100. Further, inhibitors of the active site residues including PMSF and DEPC alone were unable to inhibit enzyme activity, while cumulative presence of calcium and inhibitors reduces enzyme activity to 90%, indicating conformational changes in the protein. Molecular simulation dynamics analysis revealed a calcium binding site near the lid helix of this protein(Asn75-Ile80).
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/j.ijbiomac.2020.02.210 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
PMA1-containing extracellular vesicles of triggers immune responses and colitis progression.
Gut Microbes
December 2025
Department of Oncology, Nanjing Drum Tower Hospital, State Key Laboratory of Pharmaceutical Biotechnology, Affiliated Hospital of Medical School, Nanjing University, Nanjing, China.
() exhibits aberrant changes in patients with colitis, and it has been reported to dominate the colonic mucosal immune response. Here, we found that PMA1 expression was significantly increased in from patients with IBD compared to that in healthy controls. A Crispr-Cas9-based fungal strain editing system was then used to knock out PMA1 expression in .
View Article and Find Full Text PDFActivity-Based Bioluminescent Logic-Gate Probe Reveals Crosstalk Between the Inflammatory Tumor Microenvironment and ALDH1A1 in Cancer Cells.
JACS Au
January 2025
Department of Chemistry, Beckman Institute for Advanced Science and Technology, and Cancer Center at Illinois, University of Illinois Urbana-Champaign, Urbana, Illinois 61801, United States.
Cancer cells with high expression of aldehyde dehydrogenase 1A1 (ALDH1A1) are more resistant to chemotherapy, contribute to tumor progression, and are associated with poor clinical outcomes. ALDH1A1 plays a critical role in protecting cells from reactive aldehydes and, in the case of stem cells, regulates their differentiation through the retinoic acid signaling pathway. Despite the importance of this enzyme, methods to study ALDH1A1 high-expressing cancer cells in vivo remain limited.
View Article and Find Full Text PDFApplying the Sabatier Principle to Decipher the Surface-Structure-Dependent Catalysis of Different Starch Granules by Pullulanase.
JACS Au
January 2025
Applied Molecular Enzyme Chemistry, Department of Biotechnology and Biomedicine, Technical University of Denmark, DK-2800 Kongens Lyngby, Denmark.
Interfacial enzyme catalysis is widespread in both nature and industry. Granular starch is a sustainable and abundant raw material for which a rigorous correlation of the surface structure with enzymatic degradation is lacking. Here pullulanase-catalyzed debranching of 12 granular starches varying in amylopectin contents and branch chain contents and lengths is shown to present a biphasic relationship characteristic of the Sabatier principle.
View Article and Find Full Text PDFConstruction of Compartmentalized Meso/Micro Spaces in Hierarchically Porous MOFs with Long-Chain Functional Ligands Inspired by Biological Signal Amplification.
JACS Au
January 2025
Key Lab for Ultrafine Materials of Ministry of Education, School of Materials Science and Engineering, East China University of Science and Technology, Shanghai 200237, China.
The creation of spatially coupled meso-/microenvironments with biomimetic compartmentalized functionalities is of great significance to achieve efficient signal transduction and amplification. Herein, using a soft-template strategy, UiO-67-type hierarchically mesoporous metal-organic frameworks (HMMOFs) were constructed to satisfy the requirements of such an artificial system. The key to the successful synthesis of HMUiO-67 is rooted in the utilization of the preformed cerium-oxo clusters as metal precursors, aligning the growth of MOF crystals with the mild conditions required for the self-assembly of the soft template.
View Article and Find Full Text PDF. Extract Attenuates Inflammation and Oxidative Damage in Cisplatin-Induced Cardiotoxicity in Rats by Promoting M2 Macrophage Polarization.
Mediators Inflamm
January 2025
Faculty of Medical Sciences, Department of Genetics, University of Kragujevac, Kragujevac, Serbia.
L. fruits and leaf extracts have a broad range of immunomodulatory, anti-inflammatory, and antioxidant effects; however, their effects on cardiac protection have not been investigated. The study aims to test the biological activity of L.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!