SusChemE 2015
International Conference on Sustainable Chemistry & Engineering
October 8-9, 2015, Hotel Lalit, Mumbai
Enzymatic hydrolysis of natural proteins for peptide production
Vaze Rutuja G1, Odaneth Annamma A1 and Lali Arvind M1
1
DBT-ICT-Centre for Energy Biosciences, Institute of Chemical Technology (University under section 3 of UGCact-1956, Elite
Status and Centre of Excellence-Govt. of Maharashtra, TEQIP Phase II funded), Matunga, Mumbai400019, India
E-mail addresses: rutujagvaze@gmail.com , a.dbtceb@gamail.com , arvindmlali@gmail.com
1. Introduction:
Peptides are formed by de-polymerization of proteins which can be done using acid hydrolysis. Conventionally, mild acid
conditions are used for peptide formation while strong acids under high temperature yield complete hydrolysis of proteins to
amino acids. Demerits of this process are high temperature, longer time and degradation of some amino acids; however enzymes
can be used instead of acids which will make hydrolysis process greener and sustainable [1].
Though proteins are made up of same 20 amino acids, change in amino acid composition reflects in their functional properties
[2]. Thus different proteins would be hydrolyzed using immobilized protease to obtain protein hydrolysates. Here the bioactivity
of hydrolysate is the function of degree of hydrolysis. The higher degree of hydrolysis will produce low molecular weight
peptides, some peptides have medicinal properties like cell signaling or can act as drugs to treat the disease; other ones can be
used in cosmetic preparation due to their anti-oxidant activity. Thus we can use protein hydrolysates or peptides generated
thereof for different purpose depending upon their functionality [3]. Our aim is to produce protein hydrolysates from different
proteins and check their efficacy in various applications.
2. Material and Methods:
Immobilized protease is used for protease hydrolysis. The reaction in carried out in batch mode at 50°C. Different proteins such
as soy protein, bovine serum albumin and whey proteins are used as substrates which are dissolved in suitable buffer at
25mg/ml concentration. The hydrolysis is carried out for 3-5hrs. Degree of hydrolysis was estimated using TNBS method.
Effect of degree of hydrolysis on molecular weight of peptides would be analyzed by GPC.
3. Significant Results and Discussion
Variation in substrate resulted in change in rate of hydrolysis. It was observed that, whey proteins were easily hydrolyzed in
comparison to two other proteins i.e. soy proteins and bovine serum albumin. Initially the E:S ratio was maintained at 1:10.
Substrate concentration had less effect on the degree of hydrolysis, but GPC analysis revealed differential molecular weight
distribution in resulting hydrolysate.
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Figure:
Following figure shows hydrolysis pattern of different proteins with progress of reaction
40
Degree of hydrolysis (%)
35
30
25
20
15
10
5
0
0
50
BSA
100
150
Time (min)
Whey concentrate
200
250
Soy Protein Isolate
Figure 1: Hydrolysis of different proteins using immobilized protease
4. Conclusions:
Attempt was made to hydrolyze different natural proteins viz. soy protein, bovine serum albumin and whey proteins using
immobilized protease. Whey proteins were readily hydrolyzed with degree of hydrolysis 35.1% followed by soy proteins
(DH=19.9%) and bovine serum albumin (DH=18.7%). Higher hydrolysis of whey proteins could be attributed to their small size
and less complex structure as compared to other two proteins. Whey proteins formed low molecular weight peptides after
hydrolysis, thus it can be concluded that, nature of native proteins plays significant role and is one of the limiting factor for
governing proteolysis. Work is being carried out to find whether presence of one or more amino acids affects the overall
hydrolysis reaction. These protein hydrolysates would be tested for their functionality and used in nutraceuticals, cell culture,
cosmetics or medicinal applications.
References
[1] O. L. Tavano, J Molecular Catalysis B: Enzymatic, 90, 2013, 1-11.
[2] J. Y. Nehete, R. S. Bhambar, M. R. Narkhede and S. R. Gawali, Pharmacogn Rev, 7(14) 2013, 107–116.
[3] H. Korhonen and A. Pihlanto, Int Dairy Journal, 16, 2006, 945-960.
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