Skip to content
Research Article Open access CC BY 4.0

Characterization of Elastolytic Protease, Asnilase from Aspergillus nidulans and Its Cytotoxicity on Human Endothelial and Epithelial Cells

Yumiko Komori, Yoshiyuki Okumura, Kazuhito Kamiya, Kenji Ogawa, Toshiaki Nikai

International Journal of Biochemistry Research & Review · pp. 1–8 · Published 7 May 2016

10.9734/IJBCRR/2016/26206

Abstract

Aims: To elucidate the pathogenicity of Aspergillus nidulans, elastolytic protease was isolated from the culture supernatant of the isolate, and its biological activity and cytotoxicity were examined. Methodology: A. nidulans (NBRC 4340) spores were cultured on YCB-elastin medium for three days and elastolytic protease was isolated from the culture supernatant by DEAE-Cellulose anion exchange chromatography. Elastolytic activity was measured by using GAAPLNA (Glutaryl-L-alanyl-L-alanyl-L-prolyl-L-leucine p-nitroanilide) as the substrate. Molecular mass and isoelectric point were determined by polyacrylamide gel electrophoresis. Substrate specificities were measured by using fibrinogen, collagen, and oxidized insulin B chain. Cytotoxicity of Asnilase on human pulmonary artery endothelial cells (HPAEC), human small airway epithelial cells (HSAEC), human bronchial/tracheal epithelial cells (HBTEC) and human pulmonary alveolar epithelial cells (HPAEpiC) was determined with colorimetric method. Results: An elastolytic protease isolated from the culture supernatant of A. nidulans was found to be homogeneous as indicated by a single band after polyacrylamide gel electrophoresis (PAGE), and the final preparation named as Asnilase. Molecular mass of this enzyme was determined to be 33,800 Da and the isoelectric point was 4.2. Asnilase cleaved the Aα and Bβ chains of fibrinogen, collagen and hydrolyzed His(5)-Leu(6) and Glu(13)-Ala(14) bonds of oxidized insulin B chain. Cytotoxic effects for various human pulmonary cells were observed when Asnilase was added at concentration of 2.0-8.0 mg/mL. Conclusion: It is shown in our current investigation that Asnilase is a newly isolated elastolytic protease from A. nidulans. It is responsible for pulmonary aspergillus infection.

Aspergillus nidulans elastolytic protease proteolytic specificity cytotoxicity

References (29)

  1. 1 A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding [DOI]
  2. 2 The Reliability of Molecular Weight Determinations by Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis [DOI]
  3. 3 Invasive Aspergillosis Disease Spectrum, Treatment Practices, and Outcomes [DOI]
  4. 4 A water-soluble tetrazolium salt useful for colorimetric cell viability assay [DOI]
  5. 5 A COLORIMETRIC METHOD FOR THE DETERMINATION OF THE PROTEOLYTIC ACTIVITY OF DUODENAL JUICE [DOI]
  6. 6 A highly water-soluble disulfonated tetrazolium salt as a chromogenic indicator for NADH as well as cell viability [DOI]
  7. 7 Purification and characterization of a protease from solid state cultures of Aspergillus parasiticus [DOI]
  8. 8 Correlation of elastase production by some strains of Aspergillus fumigatus with ability to cause pulmonary invasive aspergillosis in mice [DOI]
  9. 9 Evidence for possible involvement of an elastolytic serine protease in aspergillosis [DOI]
  10. 10 Purification and characterisation of an extracellular serine proteinase from Aspergillus fumigatus and its detection in tissue [DOI]
  11. 11 Isoation and characterisation of an extracellular alkaline protease of Aspergillus fumigatus [DOI]
  12. 12 Elastin and the lung. [DOI]
  13. 13 Purification, characterization and crystallization of an extracellular alkaline protease from Aspergillus nidulans HA‐10 [DOI]
  14. 14 Purification and characterization of a serine protease extracellularly produced by Aspergillus fumigatus in a shrimp and crab shell powder medium [DOI]
  15. 15 Elastase production in clinical isolates of Aspergillus [DOI]
  16. 16 Purification and Characterization of an Extracellular Alkaline Serine Protease from Aspergillus terreus (IJIRA 6.2) [DOI]
  17. 17 Purification and biochemical characterization of a broad substrate specificity thermostable alkaline protease from Aspergillus nidulans [DOI]
  18. 18 PepJ is a new extracellular proteinase of Aspergillus nidulans [DOI]
  19. 19 Purification and properties of the elastase from Aspergillus fumigatus [DOI]
  20. 20 Substrate specificity of a hemorrhagic proteinase from timber rattlesnake venom [DOI]
  21. 21 Isolation and characterization of an elastinolytic proteinase from Aspergillus flavus [DOI]
  22. 22 Expression of elastinolytic activity among isolates inAspergillus SectionFlavi [DOI]
  23. 23 Biochemical properties and primary structure of elastase inhibitor AFUEI from Aspergillus fumigatus [DOI]
  24. 24 The Role of Virulence Factor of Elastolytic Proteinase from Aspergillus fumigatus. [DOI]
  25. 25 Evaluation of Strategies to Improve the Production of Alkaline Protease PrtA from Aspergillus nidulans [DOI]
  26. 26 Isolation and Characterization of ElastolyticProteinase from Aspergillus fumigatus. [DOI]
  27. 27 Purification and Properties of Two Neutral Proteinases from Aspergillus nidulans [DOI]
  28. 28 Characterization of Elastolytic Proteinase from Aspergillus flavus : Comparison of Biochemical Properties to the Elastolytic Proteinase from Aspergillus fumigatus. [DOI]
  29. 29 Elastase from Aspergillus fumigatus. Its Pathogenetic Role and Inhibitor. [DOI]

Cited by 4

Studies on Elastase and Elastase Inhibitor from <i>Aspergillus flavus</i>

Toshiaki Nikai · YAKUGAKU ZASSHI · 2021

Biochemical and cellular activity of chemically synthesized elastase inhibitor (S-AFUEI) from Aspergillus fumigatus

Y. Fukui, Y. Okumura, K. Uchiya · Journal de Mycologie Médicale · 2019

Showing 2 of 4 known citations — external sources report more than can currently be individually listed.

Related research

Article metrics

Real usage data collected on this platform.

21

Page views

0

PDF downloads

2

Outbound clicks

4

Citations

Views over time

Views by country

Approximate, from request IP at view time — not citizenship or institution. Countries with fewer than 5 views are grouped as "Other".

Traffic sources

Referring site, by host.

Views and downloads exclude known bots/crawlers. Citations combines this platform's own DOI-resolved index with each external source's own reported total — see Cited by above for individually listed citing works. Last refreshed 0 seconds ago.