KARAKTERISASI ENZIM KASARCGTase- DARI ISOLAT Bacillus sp. BGG-1 BERDASARKAN PARAMETER SUHU DAN pH.
ABSTRAK
Siklodekstrin-β merupakan senyawa yang bernilai tinggi dan memiliki peran
penting dalam industri Farmasi, diantaranya sebagai stabilisator, senyawa
emulsifier, penghilang bau, dan pengubah viskositas serbuk (Mori et al., 1994).
Siklodestrin-β dapat dihasilkan dari pati dengan bantuan enzim β-Cyclodextrin
Glucosyl Transferase (CGTase-β) yang dapat diperoleh dari bakteri. Berdasarkan
penelitian sebelumnya, enzim kasar CGTase-β dapat dihasilkan oleh bakteri yang
berasal dari tanah di Jatinangor yang diberi nama Bacillus sp. BGG-1. Penelitian
ini dilakukan untuk mempelajari beberapa karakter dari enzim kasar CGTase-β,
terutama suhu dan pH optimum , serta stabilitas enzim tersebut pada berbagai
suhu dan pH. Enzim kasar diperoleh dengan cara mengekstraksi enzim CGTase-β
dari isolat Bacillus sp. BGG-1 dalam suatu media produksi, dan keberadaan
siklodekstrin-β diuji secara kualitatif dalam media Horikoshi. Pengujian optimasi
dan stabilitas suhu dilakukan dengan menginkubasi enzim dan pati dalam buffer
fosfat pada variasi suhu, sedangkan untuk optimasi dan stabilitas pH dilakukan
dengan menginkubasi enzim dan pati dalam buffer dengan variasi pH pada suhu
optimum. Pada penelitian ini juga dilakukan pengujian untuk menentukan waktu
optimum aktivitas enzim CGTase-β dari isolat Bacillus sp. BGG-1. Optimasi
waktu dilakukan dengan menginkubasi enzim dan pati dalam buffer pH dan suhu
optimum dengan variasi waktu. Pengujian aktivitas pembentukan siklodekstrin-β
dilakukan dengan pengukuran penurunan absorbansi fenolftalein pada panjang
gelombang 550 nm (Goel and Nene, 1995). Hasil penelitian menunjukkan bahwa
persen aktivitas relatif tertinggi diperoleh pada suhu 60°C, pH 8 dan waktu
inkubasi selama 48 jam. Enzim tersebut stabil pada suhu 60°C serta rentang
pH 6–8.
Kata kunci : Siklodekstrin-β, CGTase- β, Bacillus sp. BGG-1.
iii
ABSTRACT
β-cyclodextrin is a compound that has high value and important role in
pharmaceutical industry, such as stabilizer, emulsifier, deodorizing, and powder
viscosity modifiers (Mori et al., 1994). β-cyclodextrin can be produced from
starch using β-Cyclodextrin Glucosyl Transferase (β–CGTase) enzyme, which can
be obtained from bacteria. Based on previous studies, crude β–CGTase enzyme
can be produced by bacteria taken from Jatinangor soil, named Bacillus sp. BGG1. This research was carried out to study some characteristic of the crude βCGTase enzyme, especially optimum temperature and pH, as well as enzyme
stability in variation of temperature and pH. Crude enzymes was obtained by
extracting the enzyme from Bacillus sp. BGG-1 in a production medium, and the
presence of β-cyclodextrin was tested qualitatively in Horikoshi media.
Temperature optimization and enzyme stability testing against temperature were
performed by incubating the enzyme and starch in phosphate buffer at the
variations of temperature, while pH optimization and enzyme stability testing
against pH were done by incubating the enzyme and starch in a buffer with pH
variation at optimum temperature. In this research, a test also conducted to
determine the optimum time of β-CGTase enzyme activity. Optimization of time
was performed by incubating the enzyme and starch in buffer at optimum pH and
temperature, with variation of time. β-cyclodextrin production activity can be
determined by measuring the decrease of phenolphthalein absorbance at 550 nm
(Goel and Nene, 1995). The results showed that the highest relative activity
percentage was achieved at 60°C, pH 8 and 48 hours of incubation time. The
enzyme stability was found at 60 °C and a pH range of 6-8.
Key Words : β-Cyclodextrin, β-Cyclodekxtrin Glucocyl Transferase, Bacillus sp.
BGG-1.
iv
Siklodekstrin-β merupakan senyawa yang bernilai tinggi dan memiliki peran
penting dalam industri Farmasi, diantaranya sebagai stabilisator, senyawa
emulsifier, penghilang bau, dan pengubah viskositas serbuk (Mori et al., 1994).
Siklodestrin-β dapat dihasilkan dari pati dengan bantuan enzim β-Cyclodextrin
Glucosyl Transferase (CGTase-β) yang dapat diperoleh dari bakteri. Berdasarkan
penelitian sebelumnya, enzim kasar CGTase-β dapat dihasilkan oleh bakteri yang
berasal dari tanah di Jatinangor yang diberi nama Bacillus sp. BGG-1. Penelitian
ini dilakukan untuk mempelajari beberapa karakter dari enzim kasar CGTase-β,
terutama suhu dan pH optimum , serta stabilitas enzim tersebut pada berbagai
suhu dan pH. Enzim kasar diperoleh dengan cara mengekstraksi enzim CGTase-β
dari isolat Bacillus sp. BGG-1 dalam suatu media produksi, dan keberadaan
siklodekstrin-β diuji secara kualitatif dalam media Horikoshi. Pengujian optimasi
dan stabilitas suhu dilakukan dengan menginkubasi enzim dan pati dalam buffer
fosfat pada variasi suhu, sedangkan untuk optimasi dan stabilitas pH dilakukan
dengan menginkubasi enzim dan pati dalam buffer dengan variasi pH pada suhu
optimum. Pada penelitian ini juga dilakukan pengujian untuk menentukan waktu
optimum aktivitas enzim CGTase-β dari isolat Bacillus sp. BGG-1. Optimasi
waktu dilakukan dengan menginkubasi enzim dan pati dalam buffer pH dan suhu
optimum dengan variasi waktu. Pengujian aktivitas pembentukan siklodekstrin-β
dilakukan dengan pengukuran penurunan absorbansi fenolftalein pada panjang
gelombang 550 nm (Goel and Nene, 1995). Hasil penelitian menunjukkan bahwa
persen aktivitas relatif tertinggi diperoleh pada suhu 60°C, pH 8 dan waktu
inkubasi selama 48 jam. Enzim tersebut stabil pada suhu 60°C serta rentang
pH 6–8.
Kata kunci : Siklodekstrin-β, CGTase- β, Bacillus sp. BGG-1.
iii
ABSTRACT
β-cyclodextrin is a compound that has high value and important role in
pharmaceutical industry, such as stabilizer, emulsifier, deodorizing, and powder
viscosity modifiers (Mori et al., 1994). β-cyclodextrin can be produced from
starch using β-Cyclodextrin Glucosyl Transferase (β–CGTase) enzyme, which can
be obtained from bacteria. Based on previous studies, crude β–CGTase enzyme
can be produced by bacteria taken from Jatinangor soil, named Bacillus sp. BGG1. This research was carried out to study some characteristic of the crude βCGTase enzyme, especially optimum temperature and pH, as well as enzyme
stability in variation of temperature and pH. Crude enzymes was obtained by
extracting the enzyme from Bacillus sp. BGG-1 in a production medium, and the
presence of β-cyclodextrin was tested qualitatively in Horikoshi media.
Temperature optimization and enzyme stability testing against temperature were
performed by incubating the enzyme and starch in phosphate buffer at the
variations of temperature, while pH optimization and enzyme stability testing
against pH were done by incubating the enzyme and starch in a buffer with pH
variation at optimum temperature. In this research, a test also conducted to
determine the optimum time of β-CGTase enzyme activity. Optimization of time
was performed by incubating the enzyme and starch in buffer at optimum pH and
temperature, with variation of time. β-cyclodextrin production activity can be
determined by measuring the decrease of phenolphthalein absorbance at 550 nm
(Goel and Nene, 1995). The results showed that the highest relative activity
percentage was achieved at 60°C, pH 8 and 48 hours of incubation time. The
enzyme stability was found at 60 °C and a pH range of 6-8.
Key Words : β-Cyclodextrin, β-Cyclodekxtrin Glucocyl Transferase, Bacillus sp.
BGG-1.
iv