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Lampiran 1 Komposisi media peremajaan
a. Media Yeast Malt Broth (YMB)
No Jenis Bahan Jumlah (g) per liter media
1 Glukosa 4
2 Ekstrak khamir 4
3 Ekstrak malt 10
b. Media Yeast Agar (YMA)
No Jenis Bahan Jumlah (g) per liter media
1 Glukosa 4
2 Ekstrak khamir 4
3 Ekstrak malt 10
4 Agar 15
c. Media Nutrient Broth
Digunakan media siap pakai merk Difco
d. Media Nutrient Agar
Digunakan media siap pakai merk Difco e. Media PDA (Potatoes Dextrose Agar)
No Jenis Bahan Jumlah (g) per liter media 1 Dextrose 20
2 Potatoes 300
3 Agar 15
f. Media Oat meal (Daigo Actino medium no. 3 – Wako Industr. Ltd)
No Jenis bahan Jumlah (g) per liter
Lampiran 2 Komposisi media produksi
1. Media ISP 4 ( Internasional Streptomyces Project)
No Jenis Bahan Jumlah (g) per liter media 1 Soluble Starch 10 2 CaCO3 2 3 (NH4)2SO4 2 4 K2HPO4 1 5 MgSO47H2O 1 6 NaCl 1 7 FeSO4.7H2O 0,001 8 MnCl2.7H2O 0,001 9 ZnSO4.7H20 0,001
2. Media modifikasi molase-kedelai
No Jenis Bahan Jumlah (g) per liter media
1 Molase 1% (v/v) 2 CaCO3 2 3 Urea 1 4 SP-36 1 5 KCl 1 5 MgSO47H2O 1 6 NaCl 1 7 FeSO4.7H2O 0,001 8 MnCl2.7H2O 0,001 9 ZnSO4.7H20 0,001 10 Bubuk kedelai 0.5% (g/v)
Lampiran 3 Prosedur pengujian kadar gula dalam media
1. Pembuatan larutan DNS
10 gr NaoH padat di tambahkan dalam 182 g K-Na-Tartrat dan 0.5 gr Na2SO3, larutkan dalam akuades sebanyak 500 ml. Larutan diaduk dengan menggunakan stirer tanpa pemanasan sambil ditambahkan 10 g DNS sedikit demi sedikit. Akuades ditambahkan kembali dalam larutan sebanyak 500 ml sambil dilakukan pengadukan sampai semua bahan larut.
2. Metode pengujian kadar glukosa (Miller 1959)
Pereaksi Blanko (ml) Kontrol(ml) Sampel (ml)
Akuades 1
-Media - 1
Ekstrak kasar enzim - - 1
Inkubasi selama 5 hari, 10 hari, dan 15 hari
DNS 2 2 2
Panaskan dalam penangas pada suhu 1000C selama 15 menit
Larutan didinginkan lalu diukur absorbansinya pada panjang gelombang 550 nm
3. Menghitung kadar glukosa dalam media Kadar glukosa dalam media :
(Kontrol-Blanko) - (Kadar glukosa hasil inkubasi-Blanko) Kurva Standart glukosa
Kadar Glukosa (ppm) Absorbansi (550 nm) 0 0.054 100 0.189 200 0.391 300 0.632 400 0.829 500 1.012
Kurva Standart Glukosa
y = 0.002x + 0.0213 R2= 0.9954 0 0.2 0.4 0.6 0.8 1 1.2 0 100 200 300 400 500 600 konsentrasi glukosa (ppm ) A b s o rb a n s i (5 5 0 n m )
Lampiran 4 Komposisi kimia media produksi
1. Komposisi molase (Paturau 1982)
Komponen Kisaran (%) Rata-rata (%)
Air 17-25 20 Sukrosa 30-40 35 Glukosa 4-9 7 Fruktosa 5-12 9 Gula pereduksi 1-5 3 Karbohidrat lain 2-5 4 Abu 7-15 12 Komponen nitrogen 2-6 4.5
Asam bukan nitrogen 2-6 5
Lilin, steroid, dan fosfolipid 0.1-1 0.4
2. Komposisi soybean meal (Sukmadi 1996)
Komponen Kadar komponen (%)
Protein 42.59 Nitrogen 6.81 Air 4.65 Lemak 19.74 Magnesium 0.15 Mangan 0.16 Besi 0.05 Seng 0.15 Fosfor 1.48 Kalsium 0.45