Impact of microwave-assisted heating and enzymatic treatment on functional and antioxidant

109 Pages·2017·1.29 MB·English

Purdue University
Purdue e-Pubs
Open Access Theses Theses and Dissertations
12-2016
Impact of microwave-assisted heating and
enzymatic treatment on functional and antioxidant
properties of rainbow trout (Oncorhynchus
mykiss) by-product hydrolysates
Elizabeth Bich Hang Nguyen
Purdue University
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Recommended Citation
Nguyen, Elizabeth Bich Hang, "Impact of microwave-assisted heating and enzymatic treatment on functional and antioxidant
properties of rainbow trout (Oncorhynchus mykiss) by-product hydrolysates" (2016).Open Access Theses. 884.
https://docs.lib.purdue.edu/open_access_theses/884
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Graduate School Form
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PURDUE UNIVERSITY
GRADUATE SCHOOL
Thesis/Dissertation Acceptance
This is to certify that the thesis/dissertation prepared
By ElizabethBichHangNguyen
Entitled
ImpactofMicrowave-AssistedHeatingandEnzymaticHydrolysisonFunctionalandAntioxidantPropertiesofRainbowTrout
(OncorhynchusMykiss)By-Product
MasterofScience
For the degree of
Is approved by the final examining committee:
Dr.AndreaLiceaga
Chair
Dr.FernandaSanMartin
Dr.Yuan(Brad)Kim
Dr.OwenJones
To the best of my knowledge and as understood by the student in the Thesis/Dissertation
Agreement, Publication Delay, and Certification Disclaimer (Graduate School Form 32),
this thesis/dissertation adheres to the provisions of Purdue University’s“Policy of
Integrity in Research” and the use of copyright material.
Dr.AndreaLiceaga
Approved by Major Professor(s):
Dr.CarlosCorvalan 12/7/2016
Approved by:
Head of the Departmental Graduate Program Date

IMPACT OF MICROWAVE-ASSISTED HEATING AND ENZYMATIC
TREATMENT ON FUNCTIONAL AND ANTIOXIDANT PROPERTIES OF
RAINBOW TROUT (Oncorhynchus mykiss) BY-PRODUCT HYDROLYSATES

A Thesis
Submitted to the Faculty
of
Purdue University
by
Elizabeth Bich Hang Nguyen
In Partial Fulfillment of the
Requirements for the Degree
of
Master of Science
December 2016
Purdue University
West Lafayette, Indiana
ii
For my Mother
iii
ACKNOWLEDGEMENTS
First and foremost, my sincere thanks to Dr. Andrea Liceaga for her guidance throughout
every aspect of my degree and research. Words can never express fully of how much I
appreciate the opportunity and everything you have given me to learn about protein
chemistry.

Thank you to my committee members: Dr. Fernanda San Martin, Dr. Owen Jones and Dr.
Yuan Brad Kim for all their thoughtful suggestions and advice to improve my research.

Many thanks to Bell Aquaculture™ for providing the fish by-products used in this study
and Amber Jannasch at Metabolomics Laboratory (Bindley Bioscience Center) for their
contribution to the amino acid analysis.

Thank you as well to my lab-mates in the protein chemistry lab for your friendship,
advice, and support. Without all of your advice and collaboration my research would not
be where it is today.

Lastly, thank you Connie for your friendship throughout this journey in graduate school.
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TABLE OF CONTENTS
Page
LIST OF ABBREVIATIONS ............................................................................................ vi
LIST OF TABLES ............................................................................................................ vii
LIST OF FIGURES ......................................................................................................... viii
ABSTRACT ....................................................................................................................... ix
CHAPTER 1. LITERATURE REVIEW ......................................................................... 1
1.1 Rainbow Trout Aquaculture in Indiana ..................................................................... 1
1.2 Fish Processing and By-products ............................................................................... 2
1.3 Fish Protein Hydrolysates (FPH) ............................................................................... 4
1.3.1 Enzymes used for FPH .................................................................................... 9
1.3.2 FPH Functionality .......................................................................................... 10
1.3.3 Protein solubility ............................................................................................ 12
1.3.4 Emulsifying activity ....................................................................................... 13
1.3.5 Foaming capacity .......................................................................................... 14
1.4 Antioxidant properties of FPH ................................................................................. 15
1.5 Microwave-assisted hydrolysis as an alternative method for obtaining FPH .......... 17
1.5.1 Microwave-Assisted Hydrolysis .................................................................... 18
1.6 Hypothesis and Research Objectives ....................................................................... 20
1.6.1 Hypothesis ..................................................................................................... 20
1.6.2 Objectives ...................................................................................................... 20
CHAPTER 2. MATERIALS AND METHODS ............................................................ 21
2.1 Materials .................................................................................................................. 21
2.1.1 Experiential Design ....................................................................................... 21
v

2.2 Production of Fish Protein Hydrolysates (FPH) ...................................................... 23
2.3 Proximate composition ............................................................................................ 24
2.4 Degree of Hydrolysis ............................................................................................... 24
2.5 Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis ............................... 25
2.6 Amino Acid Composition ........................................................................................ 26
2.7 Protein Solubility ..................................................................................................... 26
2.8 Emulsifying activity index (EAI)............................................................................. 27
2.9 Foaming properties .................................................................................................. 27
2.10 Surface hydrophobicity ............................................................................................ 28
2.11 Antioxidant Activity ................................................................................................ 29
2.11.1 1 2,2-Diphenyl-1-picryhydrazyl (DPPH) Radical-Scavenging Activity ..... 29
2.11.2 Ferric ion reducing antioxidant capacity (FIRC) ........................................ 29
2.12 Statistical analysis .................................................................................................... 30
CHAPTER 3. RESULTS AND DISCUSSION ............................................................. 31
3.1 Proximate Composition ........................................................................................... 31
3.2 Degree of Hydrolysis (DH) ...................................................................................... 34
3.3 SDS-PAGE .............................................................................................................. 38
3.4 Protein Solubility ..................................................................................................... 40
3.5 Surface Hydrophobicity ........................................................................................... 43
3.6 Emulsifying properties ............................................................................................. 46
3.7 Foaming Properties .................................................................................................. 48
3.8 Free Amino Acid Composition ................................................................................ 51
3.9 DPPH Radical Scavenging Activity of FPH ............................................................ 53
3.10 Ferric ion reducing antioxidant capacity (FIRC) Reducing Power ......................... 58
CHAPTER 4. CONCLUSION ....................................................................................... 60
CHAPTER 5. FUTURE DIRECTIONS ........................................................................ 61
LIST OF REFERENCES .................................................................................................. 61
APPENDIX:RESPONSE SURFACE METHOD DATA ................................................ 73
vi
LIST OF ABBREVIATIONS
FPH Fish protein hydrolysates
CH conventional heating
MW microwave heating
E:S enzyme substrate ratio
DH degree of hydrolysis
PS protein solubility
S surface hydrophobicity
o
EAI emulsifying activity index
ES emulsion stability
FC foam capacity
FS foam stability (FS),
DPPH 2,2-Diphenyl-1-picryhydrazyl radical-scavenging activity
FIRC ferric ion reducing capacity
vii
LIST OF TABLES
Table .............................................................................................................................. Page
2.1. Treatments of fish protein hydrolysates (FPH) from conventional heating (CH) and
microwave heating (MW). ................................................................................................ 22
3.1. Proximate composition of fish protein hydrolysates (FPH) from conventional heating
(CH) and microwave assisted heating (MW) with Alcalase ............................................. 33
3.2 Degree of Hydrolysis (DH) of fish protein hydrolysates (FPH) from conventional
heating (CH) and microwave assisted heating (MW) with Alcalase ................................ 37
3.3 Surface Hydrophobicity of fish protein hydrolysates (FPH) from conventional heating
(CH) and microwave assisted heating (MW) with Alcalase ............................................ 45
3.4 Free amino acid composition (%) of FPH derived from conventional heating (CH)
and microwave assisted (MW) hydrolysis with Alcalase ................................................. 52
3.5. Antioxidant properties of rainbow trout FPH produced from conventional heating
(CH) and microwave assisted (MW) hydrolysis ............................................................... 56