BAB V. KESIMPULAN DAN SARAN
5.2. Saran
Saran dari penelitian ini adalah:
1. Perlu adanya analisis molekuler isolat kapang endofit JE-DP4 yang diisolasi dari daun tanaman pepaya untuk diidentifikasi.
2. Perlu dilakukan pengujian variasi pH aktivitas enzim protease isolat kapang endofit JE-DP4 untuk mengetahui faktor lain yang memengaruhi aktivitas enzim.
3. Kapang endofit daun pepaya yang kemampuan aktivitas enzimnya menurun dapat diinduksi dengan penambahan ekstrak daun pepaya pada media pertumbuhan kapang agar kondisinya sesuai lingkungan tempat tumbuh kapang endofit.
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51 LAMPIRAN
Lampiran 1. Komposisi reagen 1. Kasein 1%
a. Kasein 0,5 g b. Akuades 50 mL 2. Asam trikloroasetat 5%
a. Asam trikloroasetat 2,5 g b. Akuades 50 mL
3. Bufer Tris-HCl 0,2 M pH 7,5
a. Tris Base (hydroxymethyl)-aminomethan 1,211 g b. Akuades 50 mL
c. HCl pekat 4. Reagen Bradford
a. Coomassie Brilliant Blue G-250 25 mg b. Metanol 50 mL
c. Asam fosfat (H3PO4) 85% 50 mL d. Akuades 250 mL
e. Disaring, ditambahkan akuades 175 mL 5. Larutan standar tirosin stok 3 mg/mL
a. L-tyrosine 120 mg b. Akuades 20 mL
6. Larutan standar Bovine Serum Albumin (BSA) stok 50 µg/mL a. BSA 0,25 mg
b. Akuabides 5 mL
Lampiran 2. Hasil pengukuran larutan dan kurva standar tirosin a. Pengukuran larutan standar tirosin
No. L-tyrosine (mg/mL)
Stok (mL)
HCl 0,1 M (mL)
Nilai Absorbansi (λ280 nm)
1. 0,01 0,01 0,99 0,119
2. 0,02 0,02 0,98 0,250
3. 0,03 0,03 0,97 0,364
4. 0,04 0,04 0,96 0,457
5. 0,05 0,05 0,95 0,548
6. 0,06 0,06 0,94 0,637
7. 0,07 0,07 0,93 0,774
8. 0,08 0,08 0,92 0,848
9. 0,09 0,09 0,91 0,939
10. 0,10 0,10 0,90 0,984
b. Kurva standar tirosin
Lampiran 3. Hasil pengukuran larutan dan kurva standar bovine serum albumin (BSA)
a. Pengukuran larutan standar BSA
No. BSA
(µg/mL)
Stok (mL)
ddH2O (mL)
Nilai Absrobansi
(λ595 nm)
1. 2 0,08 1,92 0,0217
2. 4 0,16 1,84 0,0387
3. 6 0,24 1,76 0,0531
4. 8 0,32 1,68 0,0786
5. 10 0,40 1,60 0,1000
6. 12 0,48 1,52 0,1177
7. 14 0,56 1,44 0,1321
8. 16 0,64 1,36 0,1428
9. 18 0,72 1,28 0,1680
10. 20 0,80 1,20 0,1825
b. Kurva standar BSA
Lampiran 4. Nilai pH pertumbuhan kapang endofit JE-DP4 menggunakan substrat kasein dan susu skim
No. Hari Ulangan
Substrat kasein Substrat susu skim
pH ƩpH pH ƩpH
Lampiran 5. Konsentrasi tirosin yang dilepaskan hasil hidrolisis protein
No. Hari Suhu (°C) Konsentrasi tirosin yang dilepaskan (µg/mL) Substrat kasein Substrat susu skim
1. 1 30 0 0
2 30 19,66 6,16
3 30 42,05 14,94
4 30 44,46 21,35
5 30 49,08 20,02
6 30 57,09 15,55
7 30 69,21 0
2. 1 37 127,43 0
2 37 24,33 13,50
3 37 43,23 17,25
4 37 42,41 22,08
5 37 54,27 24,85
6 37 66,44 13,81
7 37 78,10 0
3. 1 44 52,73 7,14
2 44 21,61 14,17
3 44 48,11 17,87
4 44 50,52 25,21
5 44 57,24 30,81
6 44 80,35 23
7 44 89,75 0
Lampiran 6. Peremajaan kapang endofit daun pepaya
Gambar 1. Isolat kapang endofit JE-BP1 (tampak depan)
Gambar 2. Isolat kapang endofit JE-BP1 (tampak belakang)
Gambar 3. Isolat kapang endofit JE-BP3 (tampak depan)
Gambar 4. Isolat kapang endofit JE-BP3 (tampak belakang)
Gambar 5. Isolat kapang endofit JE-DP4 (tampak depan)
Gambar 6. Isolat kapang endofit JE-DP4 (tampak belakang)
Lampiran 7. Skrining aktivitas enzim protease kapang endofit daun tanaman pepaya
a. Hari ke-4
Gambar 1. Isolat kapang endofit JE-DP4 (tampak depan)
Gambar 2. Isolat kapang endofit JE-DP4 (tampak belakang)
Gambar 3. Isolat kapang endofit JE-BP1 (tampak depan)
Gambar 4. Isolat kapang endofit JE-BP1 (tampak belakang)
Gambar 5. Isolat kapang endofit JE-BP3 (tampak depan)
Gambar 6. Isolat kapang endofit JE-BP3 (tampak belakang)
b. Hari ke-5
Gambar 1. Isolat kapang endofit JE-DP4 (tampak depan)
Gambar 2. Isolat kapang endofit JE-DP4 (tampak belakang)
Gambar 3. Isolat kapang endofit JE-BP1 (tampak depan)
Gambar 4. Isolat kapang endofit JE-BP1 (tampak belakang)
Gambar 5. Isolat kapang endofit JE-BP3 (tampak depan)
Gambar 6. Isolat kapang endofit JE-BP3 (tampak belakang)
c. Hari ke-6
Gambar 1. Isolat kapang endofit JE-DP4 (tampak depan)
Gambar 2. Isolat kapang endofit JE-DP4 (tampak belakang)
Gambar 3. Isolat kapang endofit JE-BP1 (tampak depan)
Gambar 4. Isolat kapang endofit JE-BP1 (tampak belakang)
Gambar 5. Isolat kapang endofit JE-BP3
(tampak depan) Gambar 6. Isolat kapang endofit JE-BP3 (tampak belakang)
d. Hari ke-7
Gambar 1. Isolat kapang endofit JE-DP4 (tampak depan)
Gambar 2. Isolat kapang endofit JE-DP4 (tampak belakang)
Gambar 3. Isolat kapang endofit JE-BP1 (tampak depan)
Gambar 4. Isolat kapang endofit JE-BP1 (tampak belakang)
Gambar 5. Isolat kapang endofit JE-BP3 (tampak depan)
Gambar 6. Isolat kapang endofit JE-BP3 (tampak belakang)
Lampiran 8. Hasil uji statistik normalitas data aktivitas enzim protease isolat kapang endofit JE-DP4 pada ketiga suhu terhadap waktu inkubasi menggunakan substrat kasein
Hipotesis:
H0 : Data berdistribusi normal H1 : Data berdistribusi tidak normal
Terlihat bahwa nilai probabilitas pada kolom Sig adalah 0,810; 0,821;
0,277; 0,714; 0,699; 0,785; 0,855 atau probabilitas >0,05. Dengan demikian H0 diterima atau distribusi data aktivitas enzim protease menggunakan substrat kasein pada ketiga suhu selama 7 hari inkubasi berdistribusi normal.
Lampiran 9. Hasil uji statistik homogenitas data aktivitas enzim protease isolat kapang endofit JE-DP4 pada ketiga suhu terhadap waktu inkubasi menggunakan substrat kasein
Hipotesis:
H0 : varians ketiga populasi homogen H1 : varians ketiga populasi heterogen
Terlihat bahwa nilai probabilitas pada kolom Sig adalah 0,011 di bawah 0,05 (0,011 < 0,05). Dengan demikian H0 ditolak atau varians ketiga populasi heterogen, sehingga analisis variansi satu jalur tidak bisa dilakukan. Analisis statistik data aktivitas enzim protease isolat kapang endofit JE-DP4 pada ketiga suhu terhadap waktu inkubasi menggunakan substrat kasein dilakukan menggunakan statistik non parametrik uji Kruskal-Wallis.
Lampiran 10. Hasil uji statistik Kruskal-Wallis data aktivitas enzim protease isolat kapang endofit JE-DP4 pada ketiga suhu terhadap waktu inkubasi menggunakan substrat kasein
Hipotesis:
H0 : tidak terdapat perbedaan rata-rata aktivitas enzim protease menggunakan substrat kasein dari ketiga suhu terhadap waktu inkubasi selama 7 hari
H1 : terdapat perbedaan rata-rata aktivitas enzim protease menggunakan substrat kasein dari ketiga suhu terhadap waktu inkubasi selama 7 hari
Terlihat bahwa pada kolom Asymp. Sig adalah 0,037 atau probabilitas di bawah 0,05 (0,037 < 0,05). Dengan demikian H0 ditolak atau terdapat perbedaan rata-rata aktivitas enzim protease menggunakan substrat kasein dari ketiga suhu terhadap waktu inkubasi selama 7 hari. Hasil uji Kruskal-Wallis untuk melihat perbedaan signifikannya dilakukan uji stepwise step-down.
Uji stepwise step-down menunjukkan perbedaan signifikan rata-rata aktivitas enzim pada hari ke-2 dan ke-7 karena terletak pada kolom subset yang berbeda. Pada hari ke-1, hari ke-3, hari ke-4, hari ke-5, dan hari ke-6 tidak menunjukkan perbedaan signifikan karena terletak pada kolom subset yang sama.
Rata-rata aktivitas enzim protease tertinggi dari ketiga suhu pada hari ke-7 sebesar
Rata-rata aktivitas enzim protease tertinggi dari ketiga suhu pada hari ke-7 sebesar
