September 22nd 2022

OPTIMIZATION OF ENZYMATIC HYDROLYSIS OF MACKEREL (Rastrelliger sp.)

LaksitoPROTEIN
MUSCLE Bramantyo 1
, Putut H. Riyadi
HYDROLYSATE 1
, Romadhon
USING 1,
RESPONSEIma Wijayanti
SURFACE 1
, Jasmin Naher2
METHODOLOGY
1
Department of Fisheries Product Technology
Faculty of Fisheries and Marine Science
Diponegoro University
2
Department of Fish Technology
Faculty of Fisheries
Hajee Mohammad Danesh Science and Technology University

The 7th International Conference on Tropical and Coastal Region Eco Development 2022
 Introduction
• Widely available in South East Asia mainly in Indonesia
Mackerel
• Rich in protein (20,83%) and low on fat (1,03%)

• Common method for protein extraction

Hydrolysis
• Enzyme used for this study is Flavourenzyme (4% E:S)

• Factors that influence hydrolysis have been studied (pH, time, temperature)
Optimization • Optimized variable for hydrolysis is Degree of Hydrolysis
• Optimization method used is RSM

The 7th International Conference on Tropical and Coastal Region Eco Development 2022
 RSM Design
Factor Name Units Minimum Maximum Mean Std. Dev.
A pH 6.00 8.00 7.00 0.7071
B Temperature Celsius 45.00 65.00 55.00 7.07
C Time Hour(s) 0.5000 1.50 1.0000 0.3536
Design Type Box-Behnken Runs 17.00

The 7th International Conference on Tropical and Coastal Region Eco Development 2022
 Actual Design & Analysis
Factor 1 Factor 2 Factor 3 Response 1
Run A:pH B:Temperature C:Waktu Average DH Fit Summary
Celsius Hour(s) % Sequential p- Lack of Fit p-
1 7 65 1.5 17.3737 Source value value Adjusted R² Predicted R²
2 6 65 1 14.9129
3 7 55 1 17.7293 Linear 0.8217 < 0.0001 -0.1500 -0.5995
4 8 45 1 15.5132
5 8 65 1 14.7372 2FI 0.8770 < 0.0001 -0.4005 -2.0160
6 7 45 1.5 16.8132
7 6 55 0.5 14.4912 Quadratic < 0.0001 0.0606 0.9758 0.8589 Suggested
8 6 55 1.5 15.5192
9 8 55 0.5 15.3812 Cubic 0.0606 0.9921 Aliased
10 6 45 1 13.3292
11 7 45 0.5 16.2313
12 7 55 1 17.5112
13 7 65 0.5 16.6717
14 7 55 1 17.6681
15 7 55 1 17.4321
16 8 55 1.5 15.2813
17 7 55 1 17.6123

The 7th International Conference on Tropical and Coastal Region Eco Development 2022
 Source Sum of
Squares
df Mean
Square
F-value p-value
Anova
Fit Statistics
Model 28.72 9 3.19 72.63 < 0.0001 significant
Std. Dev. 0.2096 R² 0.9894
A-pH 0.8847 1 0.8847 20.14 0.0028
Mean 16.13 Adjusted R² 0.9758
B-Temperature 0.4089 1 0.4089 9.31 0.0186
C.V. % 1.30 Predicted R² 0.8589
C-Time 0.6116 1 0.6116 13.92 0.0073
Adeq Precision 25.6032
AB 1.39 1 1.39 31.68 0.0008
AC 0.3180 1 0.3180 7.24 0.0311
BC 0.0036 1 0.0036 0.0821 0.7828 Note : The Predicted R² of 0.8589 is in reasonable agreement with the Adjusted
R² of 0.9758; i.e. the difference is less than 0.2.
A² 22.00 1 22.00 500.75 < 0.0001
B² 1.96 1 1.96 44.52 0.0003
C² 0.0785 1 0.0785 1.79 0.2232
Residual 0.3076 7 0.0439
not
Lack of Fit 0.2504 3 0.0835 5.84 0.0606 significant
Pure Error 0.0571 4 0.0143
Cor Total 29.03 16

Note : Characters colored in red show significant model terms (P Values < 0.0500)

The 7th International Conference on Tropical and Coastal Region Eco Development 2022
 Equations
Average DH = Average DH =
+17.59 -146.03606
+36.14329 pH
+0.3326 A
+1.17934 Temperatur
+0.2261 B
+5.26244 Waktu
+0.2765 C
-0.058993 pH * Temperatur
-0.5899 AB -0.563950 pH * Waktu
-0.2820 AC Temperatur *
+0.006005
Waktu
+0.0300 BC
-2.28587 pH²
-2.29 A²
-0.006816 Temperatur²
-0.6816 B²
-0.546030 Waktu²
-0.1365 C²

Coded Equation Actual Equation

The 7th International Conference on Tropical and Coastal Region Eco Development 2022
 Model Graphs

Term AB (PH & Temperature) Term AC (PH & Time)

The 7th International Conference on Tropical and Coastal Region Eco Development 2022
 Changes in Amino Acid
Mackerel Protein
Amino Acid Mackerel Meat (mg/kg) Hydrolysate (mg/kg) Increase (mg/kg) Increase %

L-Asam Glutamat 24437,16 29269,26 4832,104 19,77%

L-Fenialanin 6122,58 6286,626 164,0456 2,68%
L-Valin 9095,69 9142,791 47,10131 0,52%
L-Alanin 9974,98 11391,03 1416,048 14,20%
Glisin 7336,14 8146,372 810,2318 11,04%
L-Asam Aspartat 15584,36 17364,33 1779,967 11,42%
L-Leusin 13647,97 14013,46 365,4892 2,68%
L-Prolin 4830,82 5642,474 811,6543 16,80%
L-Histidin 6019,67 7648,469 1628,799 27,06%

The 7th International Conference on Tropical and Coastal Region Eco Development 2022