ResearchArticle
Effects of Tormentic Acid and the Extracts from Callistemon
citrinus on the Production of Extracellular Proteases by
Staphylococcus aureus
Rumbidzai Mashezha, Molly Mombeshora, and Stanley Mukanganyama
Department of Biochemistry, University of Zimbabwe, Harare, Zimbabwe
Correspondence should be addressed to Stanley Mukanganyama; smukanganyama@medic.uz.ac.zw
Received 16 September 2019; Revised 6 March 2020; Accepted 16 March 2020; Published 21 April 2020
Academic Editor: Saad Tayyab
Copyright © 2020 Rumbidzai Mashezha et al. is is an open access article distributed under the Creative Commons Attribution
License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is
properly cited.
Staphylococcus aureus is among the common nosocomial pathogens. Antibiotics have been used to treat S. aureus infections.
However, there has been increased mortality associated with drug-resistant strains of S.aureus. Extracellular proteases have been
implicated to be responsible for the transition of S.aureus from an adhesive pathogen to an invasive pathogen. e development of
resistant strains has necessitated the search for new sources of drugs. Plants have been traditionally used as sources of therapeutic
molecules. e objective of this study was to determine the effect of tormentic acid and the extracts from Callistemoncitrinus on
the production of extracellular proteases by S. aureus. e broth microdilution antibacterial susceptibility assay was used to
determine the antibacterial effects of tormentic acid and the extracts on S.aureus. Both extracts showed a minimum inhibitory
concentration (MIC) value of 50 μg/ml. e water : ethanol (50 : 50) and the dichloromethane : methanol (50 : 50) extracts were
found to be bactericidal against S. aureus at a concentration of 100 μg/ml and 50 μg/ml, respectively. e effect of tormentic acid
and extracts on extracellular protease production was investigated using the protease assay. A zone of proteolytic activity (Pr) was
measured as the ratio of the diameter of the colony to the total diameter of colony plus zone of hydrolysis. e extracts reduced the
production of extracellular proteases, while tormentic acid completely inhibited the production of extracellular proteases by S.
aureus. e Pr value for tormentic acid was found to be 1. e Pr values of the dichloromethane : methanol extract and the water :
ethanol extract were 0.92 and 0.84, respectively. In conclusion, tormentic acid was shown to inhibit extracellular protease
production; therefore, there is need to explore its use in antivirulence therapy to combat S. aureus infections.
1. Introduction
ere has been an increase in severe Staphylococcus aureus
infections caused by drug-resistant strains [1]. is increase
in infections has been partly attributed to the hypersecretion
of proteases by the bacteria [2]. Extracellular proteases are
used by the bacteria to resist antibiotics, for survival during
interaction with the innate immune system of host and
formation of skin abscess and as effectors of virulence-de-
terminant stability. Studies have shown that S.aureus strains
which lack protease genes have decreased the abscess for-
mation and impaired organ invasion [3]. Extracellular
proteases are required for growth of bacteria in peptide-rich
environments, serum, and blood and in the presence of
antimicrobial peptides. Extracellular proteases increase the
invasiveness of the bacteria into the body as they cleave
proteins such as elastin. e cleaving of elastin allows the
bacteria to enter into organs and the bloodstream allowing
the bacteria to spread to different parts of the body. Ex-
tracellular proteases also assist in resisting phagocytosis by
human leukocytes [4]. Proteases such as aureolysin have the
ability to cleave the human cathelicin, therefore, conferring
the bacteria’s resistance to this protein [1].
e mechanism by which S. aureus cells control the
production of virulence factors is through the action of
secreted proteases [4]. Extracellular proteases are produced
alongside toxins and other exoenzymes to control their
stability [5]. e resulting stability facilitates the coordinated
Hindawi
Biochemistry Research International
Volume 2020, Article ID 6926320, 6 pages
https://doi.org/10.1155/2020/6926320