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Sains Malaysiana 50(3)(2021): 735-742
http://doi.org/10.17576/jsm-2021-5003-15
Evaluation of Chitin as a Biomarker of Pathogenic Fungal
Isolates
(Penilaian
Kitin sebagai Biopenanda Pencilan Kulat Patogen)
AHMAD SHEHAB AHMAD LAFI1,2,
JACINTA SANTHANAM3*, TZAR MOHD NIZAM KHAITHIR4, NUR
FASHYA MUSA5 & FAHRUL HUYOP1
1Department of Biotechnology and
Medical Engineering, Faculty of Biosciences and Medical Engineering, Universiti
Teknologi Malaysia, 81310 Johor Bahru, Johor Darul Takzim, Malaysia
2Center of Desert Studies, University
of Anbar, Ramadi, Iraq
3Biomedical Science Programme, Universiti
Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Federal
Territory, Malaysia
4Department of Medical Microbiology and Immunology, Faculty
of Medicine, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaacob
Latiff, Bandar Tun Razak Cheras, 56000 Kuala Lumpur, Federal Territory,
Malaysia
5Institute of Bioproduct Development, Universiti Teknologi
Malaysia, 81310 Johor Bahru, Johor Darul Takzim, Malaysia
Received: 28 May 2020/Accepted: 27
August 2020
ABSTRACT
Chitin is a polysaccharide component of the inner cell wall
of fungi that has been used to estimate fungal invasion in plant products.
However, its detection in major pathogenic fungal species has not been
investigated. The present study aimed to determine the chitin contents of
pathogenic fungal species in order to evaluate its diagnostic potential as a
biomarker for fungal infections. High performance liquid chromatography (HPLC)
was used to measure chitin content. Pure chitin was acid hydrolyzed and the
fluorescence of 9-fluorenylmethylchloroformate (FMOC-CI) derivatives of
glucosamine produced were measured. The chitin contents of ten pathogenic
fungal isolates were determined per mycelial dry weight. They varied from 18.61
(± 0.09) to 47.12 (± 0.50) µg/mg dry mycelial weight. Candida albicans and Cryptococcus
neoformans exhibited the highest and
lowest levels of chitin, respectively. Based upon relative amounts of chitin
produced, three groups namely: high (Candida albicans, Cryptococcus gattii,
Aspergillus niger and Penicillium at 47.12, 46.98, 46.05, and 44.15 µg/mg
respectively), medium (Rhizopus, Aspergillus fumigatus, Fusarium solani, and Mucor at 36.61, 36.30, 35.03, and 34.84 µg/mg, respectively), and low (Candida
tropicalis and Cryptococcus
neoformans at 20.78 and 18.61 µg/mg,
respectively), were identified. Chitin was not detectable in bacterial isolates
used as controls. The chitin detection method offers a sensitive and specific
tool for the quantification of chitin in pathogenic fungal isolates. The
detection of chitin may be a useful assay for the diagnosis of fungal
infections in clinical samples.
Keywords: Biomarker; chitin; diagnosis; HPLC; pathogenic
fungi
ABSTRAK
Kitin adalah komponen polisakarida pada dinding sebelah dalam kulat yang telah digunakan untuk menentukan kehadirankulat pada hasil tumbuhan. Walau bagaimanapun, pengesanan kitin dalam kulatpatogen yang utama belum lagi dikaji. Kajian ini bertujuan menentukan kandungan kitin dalam spesieskulat patogen untuk menilai potensi diagnostiknya sebagai biopenanda jangkitan kulat. Kaedah kromatografi cecair prestasi tinggi (HPLC) telah digunakan untuk mengukur kandungan kitin. Hidrolisasi asid dilakukan terhadap kitin tulen untuk menghasilkan terbitan glukosamin 9-fluorenilmetilkloroformat (FMOC-CI) yang berpendafluor, lalu diukur. Kandungan kitin bagi sepuluh pencilan kulat patogen ditentukan berdasarkan berat kering miselia. Hasil yang diperoleh adalah dalam julat 18.61 (± 0.09) ke 47.12 (± 0.50) µg/mg berat kering miselia. Kandungan tertinggi kitin terdapat pada Candida albicans manakala kandungan yang paling rendah pada Cryptococcus neoformans. Berdasarkan kandungan relatif kitin, tiga kumpulan dikenal pasti, iaitu tinggi (Candida albicans, Cryptococcus gattii, Aspergillus niger dan Penicillium dengan 47.12, 46.98, 46.05 dan 44.15 µg/mg masing-masing), sederhana (Rhizopus,
Aspergillus fumigatus, Fusarium solani dan Mucor dengan 36.61, 36.30, 35.03 dan 34.84 µg/mg masing-masing) dan rendah (Candida
tropicalis dan Cryptococcus
neoformans dengan 20.78 dan 18.61 µg/mg masing-masing). Kitin tidak dapat dikesan pada pencilan bakteria yang diguna sebagai kawalan. Kaedah pengesanan kitin boleh digunakan sebagai suatu alat khusus dan sensitif untuk kuantifikasi kitin pada pencilan kulat patogen. Pengesanan kitin merupakan suatu asai yang mungkin berguna untuk diagnosis jangkitan kulat dalam sampel klinikal.
Kata kunci: Biopenanda; diagnosis; HPLC;
kitin; kulat patogen
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*Corresponding author; email: jacinta@ukm.edu.my
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