LAMPIRAN
Lampiran 1. Tabel cara kerja Pengukuran Aktivitas Protease Digesti VOLUME (Ml) PEREAKSI
Standard
Sampel
Kontrol
Balanko
Tirosin
0.50
-
-
-
Kasein 5%
-
0.50
-
-
Buffer
0.50
0.50
0.50
0.50
Ekstrak
-
0.25
-
-
Akuades
0.25
-
0.75
0.75
TCA
-
-
-
-
standard
enzim
Di Inkubasi selama 60 menit pada suhu 370c TCA
0.75
0.75
-
0.75
Kasein 1%
-
-
0.50
-
Di masukkan semua tabung kedalama refrigerator selama minimal 1 jam Sentrifugasi pada kecepatan 6000 rpm selama 15 menit Ukur absorbansinya pada panjang gelombang 280 nm
Lampiran 2. Tabel Cara Kerja Pengukuran Aktivitas Amilase Digesti VOLUME (Ml) PEREAKSI
Standard
Sampel
Kontrol
Balanko
Maltosa
0.75
-
-
-
Amilum 1%
-
0.75
-
-
Ekstrak enzim
-
0.25
-
-
Akuades
0.25
-
0.25
1.0
DNS
-
-
0.50
-
standard
Inkubasi selama 30 menit pada suhu 370c DNS
0.50
0.50
-
0.50
Amilum 1%
-
-
0.75
-
Di masukkan semua tabung kedalama refrigerator selama 5 menit Setelah dingin di tambahkan akuades sebanyak 1.5 ml Ukur absorbansinya pada panjang gelombang 540 nm
Lampiran 3. Tabel cara kerja Penentuan Kadar Protein Supernatan VOLUME (Ml) PEREAKSI
Standard
Sampel
Kontrol
Balanko
Kasein
0.1
-
-
-
Ekstrak enzim
-
0.1
-
-
Pereaksi C
5.0
-
-
-
standard
Inkubasi selama 30 menit pada suhu 370c Pereaksi D
0.5
0.5
-
-
Di kocok segera dan biarkan pada suhu kamar selama paling sedikit 10 menit Ukur absorbansinyapada panjang gelombang 660 nm Kadar protein sampel di tentukan dengan kurva standard
Lampiran 4. Analisis Aktivitas Protease pada Hepatopankreas dan Intestine Univariate Analysis of Variance Descriptive Statistics Dependent Variable: Protease mmol/mg/menit (U/menit) Ukuran B
S
K
Total
Tempat H N Total H N Total H N Total H N Total
Mean .58250 .78600
Std. Deviation .188951 .039758
N
.68425 .77750 .93125
.166765 .078122 .073355
8 4 4
.85438 .60675 .84850
.108054 .090614 .293229
8 4 4
.72763 .65558 .85525 .75542
.238887 .147835 .170907 .186607
8 12 12 24
4 4
Tests of Between-Subjects Effects Dependent Variable: Protease mmol/mg/menit (U/menit) Source Corrected Model Intercept Ukuran Tempat Ukuran * Tempat Error Total Corrected Total
Type III Sum of Squares .372a 13.696 .125
df 5 1 2
.239 .008 .429 14.497
Mean Square .074 13.696 .063
F 3.123 574.802 2.624
Sig. .033 .000 .100
.239 .004 .024
10.039 .163
.005 .850
1 2 18 24
.801 23 a. R Squared = .465 (Adjusted R Squared = .316)
Post Hoc Tests Estimates Dependent Variable: Protease mmol/mg/menit (U/menit) Tempat H N
Mean .656
Std. Error .045
.855
.045
95% Confidence Interval Lower Bound Upper Bound .562 .749 .762
.949
Pairwise Comparisons Dependent Variable: Protease mmol/mg/menit (U/menit)
(I) Tempat H N
(J) Tempat N H
Mean Difference (I-J) Std. Error -.200* .063 .200* .063
a
Sig. .005 .005
95% Confidence Interval for a Difference Lower Bound Upper Bound -.332 -.067 .067 .332
Based on estimated marginal means *. The mean difference is significant at the .05 level. a. Adjustment for multiple comparisons: Least Significant Difference (equivalent to no adjustments).
Univariate Tests Dependent Variable: Protease mmol/mg/menit (U/menit)
Contrast Error
Sum of Squares .239
df
.429
1
Mean Square .239
18
.024
F 10.039
Sig. .005
The F tests the effect of Tempat. This test is based on the linearly independent pairwise comparisons among the estimated marginal means.
3. Ukuran * Tempat Dependent Variable: Protease mmol/mg/menit (U/menit) Ukuran B S K
Tempat H N H N H N
95% Confidence Interval Lower Bound Upper Bound .420 .745
Mean .583
Std. Error .077
.786 .778 .931
.077 .077 .077
.624 .615 .769
.948 .940 1.093
.607 .848
.077 .077
.445 .686
.769 1.011
Lampiran 5. Hasil Analisis Aktivitas Amilase pada Hepatopankreas dan Intestine Univariate Analysis of Variance Between-Subjects Factors Ukuran
Tempat
1 2 3 1 2
Value Label B S K H N
N 8 8 8 12 12
Descriptive Statistics Dependent Variable: Amilase mmol/mg/menit (U/menit) Ukuran B
S
K
Total
Tempat H N Total H N Total H N Total H N Total
Mean .62225
Std. Deviation .231998
N
1.41125 1.01675 .72125
.097288 .452754 .208410
4 8 4
1.14775 .93450 .72125
.174603 .289228 .226366
4 8 4
1.17325 .94725 .68825
.230295 .321034 .207065
4 8 12
1.24408 .96617
.201794 .347242
12 24
4
Tests of Between-Subjects Effects Dependent Variable: Amilase mmol/mg/menit (U/menit) Source Corrected Model Intercept Ukuran Tempat Ukuran * Tempat Error Total Corrected Total
Type III Sum of Squares 2.049a 22.403
df 5 1
Mean Square .410 22.403
F 10.181 556.640
Sig. .000 .000
.031 1.854 .164
2 1 2
.016 1.854 .082
.390 46.057 2.034
.683 .000 .160
.724 25.177 2.773
18 24 23
.040
a. R Squared = .739 (Adjusted R Squared = .666)
Post Hoc Tests Estimates Dependent Variable: Amilase mmol/mg/menit (U/menit) Tempat H N
Mean .688
Std. Error .058
1.244
.058
95% Confidence Interval Lower Bound Upper Bound .567 .810 1.122
1.366
Pairwise Comparisons Dependent Variable: Amilase mmol/mg/menit (U/menit)
(I) Tempat H N
(J) Tempat N H
Mean Difference (I-J) Std. Error -.556* .082 .556* .082
a
Sig. .000 .000
95% Confidence Interval for a Difference Lower Bound Upper Bound -.728 -.384 .384 .728
Based on estimated marginal means *. The mean difference is significant at the .05 level. a. Adjustment for multiple comparisons: Least Significant Difference (equivalent to no adjustments).
Univariate Tests Dependent Variable: Amilase mmol/mg/menit (U/menit)
Contrast Error
Sum of Squares 1.854 .724
df 1 18
Mean Square 1.854 .040
F 46.057
Sig. .000
The F tests the effect of Tempat. This test is based on the linearly independent pairwise comparisons among the estimated marginal means. 3. Ukuran * Tempat Dependent Variable: Amilase mmol/mg/menit (U/menit) Ukuran B S K
Tempat H N H N H N
95% Confidence Interval Lower Bound Upper Bound .412 .833
Mean .622
Std. Error .100
1.411 .721 1.148
.100 .100 .100
1.201 .511 .937
1.622 .932 1.358
.721 1.173
.100 .100
.511 .963
.932 1.384
Lampiran 6. Kurva Standar Larutan Tirosin
Absorbansi
0,6
Y = a + bx
0,5 0,4 0,3 0,2 0,1 0
0
0,05
0,1
0,25
0,5
1,0
Konsentrasi (mg/ml)
540 nm
Lampiran 7. Kurva Standar Larutan Maltosa
0,01
0,05
0,10 mg/ml
0,25
Lampiran Gambar Penelitian
Gambar 1. Akuarium ikan uji
Gambar 3. Saluran digesti ikan nilem
Gambar 5. Water Heater
Gambar 2. Timbangan Mekanik
Gambar 4. Running Enzim
Gambar 6. Sentrifugase
Gambar 7. Oven
Gambar 8. Timbangan Analitik
Gambar 9. Homogenizer
Gambar 10. Spectrophotometer tipe U-3900
