Produksi, isolasi dan karakterisasi superoksida dismutase dari Spirulina platensis yang dibiakkan dalam serum lateks Production, isolation, and characterization of superoxyde dismutase from Spirulina platensis cultured on latex serum
DOI:
https://doi.org/10.22302/iribb.jur.mp.v77i1.113Keywords:
atex serum, metalo enzyme, superoxide dismutase, blue-green microalga, antioxidant]Abstract
Abstract
Spirulina platensis is a blue-green microalga
which is frequently used for food and feed
supplements and cosmetic active agent. This
microalga also produces a strong antioxidant
namely superoxide dismutase (SOD) used as
cosmetic active agent for anti aging and anti free
radicals. SOD was isolated from S. platensis cell
biomass from local isolate grown on latex serum
on semipilot (3.5 m 3 ) and pilot scale (40 m 3 )
then dried with spray drying or sun drying and
characterized. SOD was purified with sequential
two-stage sedimentation using ammonium
sulphate and fractionated in chromatographic
column containing Sephadex G 200. The
fractions were analysed to determine the activity,
cofactor metal and amino acid composition of the
antioxidant. The results showed that
sedimentation of SOD extract with 80%
ammonium sulphate produced SOD with higher
activity compared to that of SOD from
commercial S. platensis biomass. This SOD was
successfully isolated and purified. Metaloenzyme
SOD was composed of subunits with molecular
weight of 77.78; 71.74; and 19.2 kDa, which
contained nine types of amino acids with tyrosine
and lysine as the major amino acid components.
Zn was the most predominant metal on SOD, then
followed by Fe and Mn. The main subunit
cofactors consisted of Zn 72%, Fe 25%, Mn 2%,
and Cu 1%, which were different from the
small subunit that contained of Zn 55%, Mn 31%,
Fe 14%, and Cu 4%. The stability of SOD was
achieved on pH 7.5 and temperature below 25 o C.
Abstrak
Spirulina platensis adalah mikroalga hijau
biru yang banyak digunakan sebagai suplemen
pangan, pakan, dan bahan aktif kosmetika.
Mikroalga ini juga menghasilkan antioksidan
kuat yaitu superoksida dismutase (SOD), yang
merupakan bahan aktif kosmetika anti penuaan
dini dan pencegah efek radikal bebas. SOD
diisolasi dari biomassa sel S. platensis isolat lokal
yang dibiakkan dalam serum lateks skala
semipilot (3,5 m 3 ) dan pilot (40 m 3 ) serta
dikeringkan dengan cara pengeringan kabut
(spray drying) atau penjemuran untuk kemudian
dikarakterisasi. SOD dimurnikan dengan peng-
endapan bertingkat menggunakan ammonium
sulfat dan dipisahkan dengan kolom kromatografi
berisi Sephadex G 200. Hasil pemisahan
kemudian dianalisis untuk menentukan aktivitas,
logam kofaktor serta komposisi asam amino anti
oksidan tersebut. Hasil penelitian menunjukkan
bahwa pengendapan ekstrak SOD dengan
SOD lebih tinggi dari SOD asal biomassa
S. platensis komersial. SOD tersebut telah
berhasil diisolasi dan dimurnikan. Metaloenzim
SOD tersusun atas subunit dengan BM 77,78;
71,74; dan 19,2 kDa, yang mengandung
sembilan jenis asam amino dengan tirosin dan
lisin sebagai komponen asam amino utama.
Logam yang dominan pada SOD adalah Zn,
disusul kemudian Fe dan Mn. Kofaktor sub unit
besar terdiri dari Zn 72%, Fe 25%, Mn 2%, dan
Cu 1%, berbeda dengan sub unit kecil yang
mengandung Zn 55%, Mn 31%, Fe 14%, dan Cu
4%. Stabilitas SOD S. platensis dicapai pada pH
7,5 dan suhu di bawah 25 o C
ammonium sulfat 80% menghasilkan aktivitas
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