SAINS MALAYSIANA

Extraction and Antibacterial Activity of Alpinia conchigera Rhizome Extract and 1’S -1’-Acetoxychavicol Acetate (ACA) against Streptococcus pneumoniae

(Pengekstrakan dan Aktiviti Antibakteria Ekstrak Rizom Alpinia conchigera dan 1'S -1'- Asetoksikavikol Asetat (ACA) terhadap Streptococcus pneumoniae)

NOR SYAZWANI MUHAMMAD ZAHIDAN¹, NUR UMAIRAH ATIQAH SABRI¹, SITI NOOR ADNALIZAWATI ADNAN², KHALIJAH AWANG³& NORMALIZA AB MALIK^1,*

¹Department of Conservative Dentistry and Prosthodontics, Faculty of Dentistry, Universiti Sains Islam Malaysia, 55100 Kuala Lumpur, Malaysia

²Department of Basic Science, Faculty of Dentistry, Universiti Sains Islam Malaysia, 55100 Kuala Lumpur, Malaysia

³Department of Chemistry, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia

The study aimed to investigate the antibacterial effects of Alpinia conchigera rhizome extract and 1’S-1’-acetoxychavicol acetate (ACA) against Streptococcus pneumoniae strain successfully isolated from oral rinse of older adults from an old folks’ home. The grounded rhizomes of A. conchigerawere macerated in hexane solvent to yield crude hexane extract. Then, A. conchigera rhizome extract was subjected to column chromatography to isolate ACA. Both A. conchigera rhizome extract and ACA were screened for the antibacterial activity against S. pneumoniae ATCC 49619 and S. pneumoniae isolate by disc diffusion test, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and time kill assay. The highest inhibition zone recorded by A. conchigera rhizome extract (100 mg/mL) and ACA (100 mg/mL) were 36.83 ± 0.85 mm and 30.67 ± 0.94 mm, respectively against S. pneumoniae isolate. The MIC and MBC values recorded for A. conchigera rhizome extract and ACA against both S. pneumoniae ATCC 49619 and S. pneumoniae isolate ranging from 12.50 mg/mL to 50.00 mg/mL. Based on the time kill curve, both extracts with the concentration of 2 MIC showed killing properties against S. pneumoniae isolate. Besides that, the morphology of S. pneumoniae isolate treated with ACA (12.50 mg/mL) was observed under scanning electron microscope and showed structural changes such as cell wall disruption and morphological disorder. Moreover, several virulence genes of S. pneumoniae were studied using RT-qPCR. It is observed that two genes, pavA and psaA function in adherence and attachment of S. pneumoniae to the host cells were downregulated while nanA which that responsible for biofilm production was upregulated. Therefore, both A. conchigera rhizome extract and ACA showed great potential as alternative antibacterial agent against S. pneumoniae.

Keywords: Alpinia conchigera; pneumonia; rhizome extract; Streptococcus pneumoniae; 1’S-1’-acetoxychavicol acetate

Penyelidikan ini bertujuan untuk mengkaji kesan antibakteria ekstrak rizom Alpinia conchigera dan 1'S -1'-asetoksikavikol asetat (ACA) terhadap strain Streptococcus pneumoniae yang berjaya dipencilkan daripada bilasan mulut orang dewasa yang diperoleh daripada rumah orang tua. Serbuk rizom A. conchigera direndam di dalam pelarut heksana untuk menghasilkan ekstrak heksana. Kemudian, ekstrak rizom A. conchigera tersebut dikenakan kolumn kromatografi bagi memencilkan ACA. Ujian resapan cakera (DDA), kepekatan perencatan minimum (MIC), kepekatan bakterisidal minimum (MBC) dan asai kadar masa membunuh telah dijalankan untuk menentukan sifat antibakteria kedua-dua ekstrak rizom A. conchigera dan ACA terhadap S. pneumoniae ATCC 49619 dan pencilan S. pneumoniae. Berdasarkan prosedur DDA, ekstrak rizom A. conchigera (100 mg/mL) dan ACA (100 mg/mL) masing-masing merekodkan zon perencatan tertinggi iaitu 36.83 ± 0.85 mm dan 30.67 ± 0.94 mm apabila diuji terhadap pencilan S. pneumoniae. Nilai MIC dan MBC yang direkodkan oleh ekstrak rizom A. conchigera dan ACA terhadap ATCC 49619 S. pneumoniae dan pencilan S. pneumoniae adalah daripada julat 12.50 mg/mL sehingga 50.00 mg/mL. Berdasarkan lengkung kadar masa membunuh, kedua-dua ekstrak dengan kepekatan 2 MIC menunjukkan sifat bakterisidal terhadap strain S. pneumoniae. Selain itu, morfologi strain S. pneumoniae yang telah dirawat dengan ACA (12.50 mg/mL) telah diperhatikan melalui mikroskop elektron imbasan dan menunjukkan perubahan struktur seperti pemecahan dinding sel dan kerosakan morfologi. Tambahan lagi, beberapa gen virulens S. pneumoniae telah dikaji menggunakan RT- qPCR. Hasil mendapati bahawa dua gen, pavA dan psaA, yang berfungsi untuk pelekatan S. pneumoniae terhadap sel perumah dikawal atur turun, manakala gen nanA yang berfungsi untuk penghasilan biofilem telah dikawal atur naik. Oleh itu, kedua-dua ekstrak rizom A. conchigera dan ACA menunjukkan potensi yang baik sebagai agen antibakteria alternatif melawan S. pneumoniae.

Kata kunci: Alpinia conchigera; ekstrak rizom; pneumonia; Streptococcus pneumoniae; 1’S -1’-asetoksikavikol asetat

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