Introduction: Photodynamic inactivation has been developed to kill pathogenic microbes. In addition, some techniques have been introduced to minimize the biofilm resistance to antifungal properties in inhibiting cell growth. The principle of photodynamic inactivation different to antifungal drugs therapy which is resistant to biofilms. The presence of reactive oxygen species (ROS) that generating in photodynamic inactivation mechanisms can be damaging of biofilm cells and the principle of light transmission that could be penetrating in matrix layers of extracellular polymeric substance (EPS) until reaching the target cells at the base layers of biofilm. The present work aims to explore the potential of chlorophyll extract of papaya leaf as an exogenous photosensitizer to kill the Candida albicans biofilms after being activated by the laser. The potential of chlorophyll photosensitizer was evaluated based on the efficacy of inactivation C. albicans biofilm cell through a cell viability test and an organic compound test. Methods: The treatment of photoinactivation was administered to 12 groups of C. albicans biofilm for four days using the 445 nm laser and the 650 nm laser. The 445 nm and 650 nm lasers activated the chlorophyll extract of the papaya leaf (0.5 mg/L) at the same energy density. The energy density variation was determined as 5, 10, 20, 30 and 40 J/cm2 with the duration of exposure of each laser adjusted to the absorbance percentage of chlorophyll extract of the papaya leaf. Results: The absorbance percentage of chlorophyll extracts of the papaya leaf on wavelengths of 650 nm and 445 nm respectively were 22.26% and 60.29%, respectively. The most effective treated group was a group of the laser with the addition of chlorophyll, done by the 650 nm lasers with inactivation about 32% (P=0.001), while the 445 nm lasers only 25% (P=0.061). The maximum malondialdehyde levels by treatment of the laser 650 nm were (0.046±0.004) nmol/mg. Conclusion: The use of chlorophyll extract of the papaya leaf as a photosensitizer, resulted in the maximum spectrum of absorption at 414 nm and 668 nm, which produced a maximum reduction effect after photoinactivation up to 32% (with chlorophyll) and 25% (without chlorophyll). The utilization of chlorophyll extract of the papaya leaf would increase the antifungal effects with the activation by the diode laser in the biofilm of C. albicans
AbstrakKitinase merupakan enzim hidrolitik yang dapat menghidrolisis kitin pada ikatan β-1,4-glikosidiknya dengan menghasilkan derivat-derivat kitin seperti oligomer kitin yang mempunyai banyak manfaat. Penelitian ini bertujuan untuk melakukan pengembangan produksi enzim kitinase dari sumber lokal yang melimpah di alamserta murah dengan melakukan optimasi substrat dalam hal ini digunakan substrat tetes tebu (molase) dan limbah cangkang rajungan untuk produksi enzim kitinase dari Aspergillus niger. Sebelumnya, dilakukan kultivasi isolat kapang Aspergillus niger dengan membuat kurva pertumbuhan menggunakan metode masa sel kering dimana dari hasil penelitian inokulasi optimal adalah 22 jam. Pada proses produksi, diperoleh waktu fermentasi optimal adalah 52 jam dengan menentukan uji aktivitasnya menggunakan metode turbidimetri. Hasil optimasi substrat menunjukkan bahwa enzim kitinase yang maksimal diperoleh pada penambahan molase 0,5% (b/v) dengan unit aktivitas enzim 0,14726 (U/mL) dan cangkang rajungan 2% (b/v) dengan unit aktivitas enzim yang dihasilkan 0,12826 (U/mL). Kitinase dari Aspergillus niger ini mempunyai pH optimal 6 dan suhu optimal 40 o C. Kata kunci: Aspergillus niger, kitinase, cangkang rajungan, molase AbstractChitinase is a hydrolytic enzyme that hydrolyzes chitin on β-1,4-glycosidic bond and thereby producing chitin derivatives such as chitin oligomers that have multiple benefits. The purpose of this research was to develop the production of chitinase enzyme from cheap and are abundant local nature sources, by optimizations substrate in this case the substrate used molasses and crab shell waste for the production of chitinase enzyme from Aspergillus niger. Previously, isolates of Aspergillus niger cultivated by creating a growth curve using dry cell mass method which from the results of research inoculation optimal are 22 hours. In the production process, obtained the optimum fermentation time is 52 hours to determine the activity test using turbidimetry method. Result of substrate optimizations indicate that chitinase enzyme maximum by addition of molasses obtained in 0.5% (w/v) with enzyme activity units 0.14726 (U/mL) and crab shells 2% (w/v) with enzyme activity units 0.12826 (U/mL). Chitinase from Aspergillus niger has a pH optimum 6 and temperature optimum 40 o C.
Enzim kitinase banyak digunakan dalam bidang medis, makanan, bioteknologi dan lingkungan. Banyaknya kebutuhan enzim kitinase menuntut penyediannya yang murah dan melimpah dengan teknologi produksi yang sederhana. Penelitian ini bertujuan untuk isolasi mikroba kitinolitik dari cairan fermentasi sampah organik, produksi dan uji aktivitas enzim kitinase serta mengetahui karakteristik dari enzim kitinase. Isolasi mikroba telah dilakukan dengan metode spread plate. Aktivitas kitinase ditentukan secara kualitatif dengan pengukuran indeks kitinolitik dan secara kuantitatif dengan pengukuran absorbansi menggunakan spektrofotometer Uv-Vis pada panjang gelombang 660 nm berdasarkan banyaknya substrat kitin yang dihidrolisis oleh enzim kitinase. Satu dari beberapa isolat yang didapatkan, yaitu isolat A1 menunjukkan aktivitas kitinolitik tertinggi, yaitu sebesar 1,21. Hasil identifikasi mikrobiologi menunjukkan bahwa isolat A1dinyatakan sebagai Pseudomonas pseudomallei. Bakteri ini mampu menghasilkan kitinase secara optimum pada jam ke 18 waktu fermentasi, dengan penambahan molase 0,5% (b/v) dan 1% kitin (b/v) pada media produksinya. Kitinase yang dihasilkan P. pseudomallei menunjukkan aktivitas optimum pada suhu 50 °C danpH sebesar 6.
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