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DOI:10.19026/ajfst.6.153
ISSN: 2042-4868; e-ISSN: 2042-4876
© 2014 Maxwell Scientific Publication Corp.
Submitted: January 27, 2014 Accepted: February 10, 2014 Published: September 10, 2014
Research Article
Study on Hydrolysis Conditions of Flavourzyme in Soybean Polypeptide Alcalase
Hydrolysate and Soybean Polypeptide Refining Process
1
Yongsheng Ma, 1Lintong Wang, 2Xianhui Sun, 1Jianqiang Zhang, 1Junfeng Wang and 1Yue Li
1
Chemistry and Chemical Engineering and Environment Engineering College, Weifang University,
Weifang, Shandong, 261061,
2
Agricultural Engineering College, Weifang Vocation College, Weifang, Shandong, 261041, P.R. China
Abstract: Soybean protein Alcalase hydrolysate was further hydrolyzed by adopting Flavourzyme as hydrolytic
enzyme. The optimal hydrolysis conditions of Flavourzyme was that pH was 7.0 at temperature 50°C and E/S(ratio
of enzyme and substrate) was 20LAPU/g. Bitterness value was reduced to 2 after Flavourzyme hydrolysis reaction
in optimal hydrolysis conditions. The change of molecular weight distribution range from Alcalase hydrolysate to
Flavourzyme hydrolysate was not obvious. DH (Degree of hydrolysis) of soybean protein hydrolysate was increased
to 24.2% which was improved 3.5% than Alcalase hydrolysate. Protein recovery proportion was increased to 73.2%
which was improved 0.8% than Alcalase hydrolysate. Soybean polypeptide Flavourzyme hydrolysate was
decolorized with activated carbon which optimal dosage was 1.2% solution amount (w/w). Anion/cation exchange
process was used in the desalination processing of soybean polypeptide. Ratio of anion resin and cation resin was
2:3(V/V). The volume of hydrolysate processed was 5 times as the volume of anion resin. Ash content of soybean
peptide solution reduced to 2.11% (dry basis), salinity decreased by 86% after desalination processing.
Keywords: Activated carbon decolourization, alcalase hydrolysate, degree of hydrolysis, desalination process,
flavourzyme, ion exchange resin
Corresponding Author: Yongsheng Ma, Chemistry and Chemical Engineering and Environment Engineering College, Weifang
University, Weifang, Shandong, 261061, P.R. China, Tel.: 0536-8785677, 13468486899
This work is licensed under a Creative Commons Attribution 4.0 International License (URL: http://creativecommons.org/licenses/by/4.0/).
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Adv. J. Food Sci. Technol., 6(9): 1027-1032, 2014
On the other hand soybean polypeptide mixture Optimization of flavourzyme enzymolysis reaction
was decolored using activated carbon adsorption conditions: Soybean separation protein powder was
decolourization method in this study. Activated carbon first hydrolyzed by Alcalase, so Flavourzyme
adsorption method with low cost and high safety that hydrolysis conditions of the substrate concentration had
operation is simple can achieve good decolorization been determined. In order to improve the production
effect. At the same time, part of hydrophobic amino efficiency reaction time was control in 3 h inside,
acids can be adsorpted through the adsorption effect of different conditions including enzyme concentration,
active carbon, so hydrolysate taste is improved reaction temperature and pH and so on were chosen as
obviously (Pu and Liang, 2012). influence factors on soybean protein hydrolysis coarse
In addition, hydrolysate is also needed to carry on
liquid further hydrolysis to determine optimal
desalination processing to eliminate the high salinity in
order to achieve good taste of product. In this study ion conditions of Flavourzyme debitterizing to soybean
exchange resin was adopted to make desalination protein Alcalase hydrolysate.
processing to soybean protein hydrolysate. Through the
experiment desalting effect of ion exchange resin to Bitter evaluation method: Bitter evaluation standard
hydrolysate and the role in the desalination was studied. was according to reference (Pu and Liang, 2012). It was
Finished product of soybean polypeptide is general that 1 mL soybean peptide solution sample was diluted
processed into solid powder because powder is easy to to just can be tasted the bitter taste so far, diluted
save. Concentration of soybean protein solution before multiples was regarded as bitter taste value. The
hydrolysis was 8%. Concentration of the soybean average of diluted multiples was as the final bitter taste
protein hydrolysate after centrifugal separation, value.
decolorizing processing and desalination processing
was 5% or so. However, when concentration was too Activated carbon decolorizing method: Supernatant
low, drying process was hard to make, so hydrolysate fluid from centrifugal separation was preheated to
was first for concentration before drying process. 50°C, activated carbon powder which weight ratio is 1
Because the soybean peptide solution was easy to make to 2% of the supernatant was added with stirring and
Browning reaction when the solution was heated, too constant temperature. Filtration separation was made
high temperature can cause serious Browning in after stirring 60 min. Residue was washed with water
product and Browning reaction limits its application.
which amount was 10 times as activated carbon. After
Low temperature vacuum concentration was adopted to
filtration each filtrate was mixed as soybean
avoid products Browning reaction. Hydrolysate
concentration was concentrated to 35% and then drying polypeptide clear liquid after activated carbon
process was made by using the spray drying equipment. adsorption decolourization.
dosage. When activated carbon dosage achieved 1.5% cation exchange column. The pH of effluent solution
of Flavourzyme hydrolysate, soybean peptide solution was measured which was compared with the value of
had a good color as the color when the dosage of influent solution. When sample quantity was 7.5 times
activated carbon was 2.0%. However, activated carbon as column volume, cation exchange resin was saturated.
dosage was more, the adsorption amount of protein was When sample quantity was 5 times as column volume,
also more resulting in a large number of protein loss 711 anion exchange resin was saturated.
bring the economic benefit loss. So the dosage of It can be decided that the suitable loading column
activated carbon should be reduced in order to reduce quantity ratio of anion resin and cation resin was
the protein loss. 2:3(5:7.5). Loading column quantity of 732 cation
The dosage of activated carbon was adjusted as exchange resin column was 200 mL. The capacity of
1.1, 1.2, 1.3 and 1.4% of solution. Then soybean 711 anion exchange resin column was 300 mL. 1000
peptide solutions were made adsorption decolourization mL soybean protein solution after decolorizing was
processing. Their decolourization effect was compared made desalination processing in resin column. Finally
to the effect when activated carbon dosage was 1.5%. It soybean peptide solution residual in column was eluted
was found from experiment result that when the with no salt water. The result was listed in Table 2.
activated carbon dosage of solution amount was 1.2% It was can be seen from Table 2 that protein lost
good decolorizing effect could be achieved, at the same was 2.46% [counting equation was (5.37*1000-
time the color of solution was light yellow. The protein 4.85*1080) /5.37*1000] which loss rate is not high after
loss rate in solution when the activated carbon dosage ion exchange desalination processing. Conductivity of
was 1.2 was 5.7% which was much less than 8.9% solution reduced largely after desalination processing,
when the dosage was 1.5%.
because ion in solution was removed by ion exchange
resin, ion concentration decreased. It can be concluded
Desalination effect on soybean polypeptides of ion
that ideal desalting effect was achieved. From the ash
exchange: In the desalting process by using anion and
content of entrance solution and exit solution, it can be
cation exchange resin column, desalted fluid must flow
through firstly cation exchange resin column, then it seen that 86% salt was removed by anion and cation
flow through anion exchange resin column. Because if exchange resin which effect was also good.
desalted fluid which may contain Ca2+ or other cation
flow through firstly anion exchange column which may Soybean polypeptide product analysis: Soybean
contain OH-, Ca2+ or other cation and OH- ions could peptide solid powder was gained from concentration
combine to generate precipitation which can jam resin, drying of soybean peptide solution. The color of
the column efficiency was reduced or exchange peptide powder was milk white or light yellow. The
capacity was lost. powder was soluble in water and there was no
Treated fluid after flowing through cationic precipitation after dissolving. The taste of 1% aqueous
exchange column was strongly acidic due to the H+ solution of soybean peptide solid powder was light and
from cationic resin. When the pH value of effluent there was no peculiar smell or bitter. Water content of
solution and the influent solution was roughly the same, final product was 5% and protein content was up to
the ion exchange was saturated. Similarly, because the 97% (dry basis). Soybean peptide solid powder was
OH- from anion exchange resin into the solution, the pH easy soluble in acid which soluble Nitrogen Index
value of effluent solution increases, when the pH value (NSI) was 99.7%.
of effluent solution and the influent solution was
roughly the same, the anion exchange resin was also CONCLUSION
saturated.
Both loading quantity of anion column and cation Soybean protein Alcalase hydrolysate was further
column were 300 mL. The soybean peptide solution hydrolyzed by adopting Flavourzyme as hydrolytic
after 1 h activated carbon decolorizing flowed into 732 enzyme to reduce bitterness of polypeptide. Bitterness
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Adv. J. Food Sci. Technol., 6(9): 1027-1032, 2014
of soybean polypeptide was reduced distinctly after this final product was 5% and protein content was 97% (dry
further hydrolysis reaction. The optimal hydrolysis basis). The product powder was easy soluble in acid
conditions of Flavourzyme in Alcalase hydrolysate was which soluble nitrogen index (NSI) was 99.7%.
determined as that pH was 7.0 at temperature 50°C and
E/S(ratio of enzyme and substrate) was 20LAPU/g. ACKNOWLEDGMENT
Bitterness value was reduced to 2 after Flavourzyme
hydrolysis reaction for 2 h in optimal hydrolysis This study was supported by spark plan project in
conditions. Shandong province (No.2011XH06006) and Science
It can also be concluded From the HPLC diagram and technology development project in Weifang city
of soybean peptides Flavourzyme hydrolysate that the (No.20121333).
change of molecular weight distribution range from
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