ENZYMATIC PRODUCTION OF TRANS-FREE STRUCTURED MARGARINE FAT ANALOGS by ...

251 Pages·2012·3.39 MB·English

by  Andrea Montlick

ENZYMATIC PRODUCTION OF TRANS-FREE STRUCTURED MARGARINE FAT
ANALOGS

by
GARIMA PANDE
(Under the Direction of Casimir C. Akoh)
ABSTRACT

Intake of high amounts of trans fatty acids (TFAs) have been positively correlated with
increased risk of several chronic diseases. The main aim of this research project was to
produce structured lipids (SLs) suitable for formulation of trans-free margarines. The
oils/fats used were stearidonic acid (SDA) soybean (SDASO), high stearate soybean
(HSSO), cottonseed (CO), fully hydrogenated cottonseed (HCO) oils, and palm stearin
(PS). The first specific aim was to optimize the reaction conditions of substrates for
trans-free SLs production. Second specific aim was to characterize the physical and
chemical properties of SLs and compare with physical blends. Third specific aim was to
characterize the textural and sensory properties of margarines prepared with selected SLs
and compare with commercial brands. Desirable and optimal products composition were
achieved at 50 °C, 18 h, 2:1 (HSSO:SDASO) with Lipozyme TLIM, containing 15.1
mol% stearic acid and 10.5 mol% SDA. It had desirable polymorphism (β'), thermal
properties, and solid fat content (SFC, 3.2% at 10 °C and 0.1% at 33.3 °C) for
formulation of soft margarine. The margarine formulated with this SL was trans-free,
SDA-enriched with no significant difference in sensory properties. For the second
combination, desirable SL containing 12.3 mol% stearic acid and no TFA was obtained
at 50 °C, 20 h, 2:1 (PS:HSSO) with Novozym 435. This SL was suitable for stick/hard
margarine because of its high melting completion temperature (45.4 °C). In the third
combination, SL synthesized at 56 °C, 6 h, 4:1 (PS:CO), using Novozym 435, was used
to formulate hard/industrial margarine with high oxidative stability and no TFA. In the
fourth combination, SL synthesized at 65 °C, 16.5 h, 2:1 (PS:HCO) using Lipozyme
TLIM, was selected to formulate margarine. A trans-free hard margarine with high
melting completion temperature (50.1 °C) that may be suitable for puff pastries, cooking,
or baking purposes was obtained. Different types of trans-free margarines were
produced, the most prominent being SDA-containing soft, spreadable margarine. This
research resulted in the production of trans-free SLs as an alternative to partially
hydrogenated fat and can be used by the food industry to formulate trans-free foods.

INDEX WORDS: Cottonseed oil, Fully hydrogenated cottonseed oil, High stearate
soybean oil, Lipases, Lipozyme TLIM, Novozym 435, Palm stearin, Stearidonic acid
soybean oil, Structured lipid, trans Fatty acid, trans-Free margarines
ENZYMATIC PRODUCTION OF TRANS-FREE STRUCTURED MARGARINE FAT
ANALOGS

by

GARIMA PANDE
B.Sc., G.B. Pant University of Agriculture and Technology, India, 2007
M.S., The University of Georgia, 2009

A Dissertation Submitted to the Graduate Faculty of The University of Georgia in Partial
Fulfillment of the Requirements for the Degree

DOCTOR OF PHILOSOPHY

ATHENS, GEORGIA
2012
© 2012
Garima Pande
All Rights Reserved
ENZYMATIC PRODUCTION OF TRANS-FREE STRUCTURED MARGARINE FAT
ANALOGS

by

GARIMA PANDE

Major Professor: Casimir C. Akoh
Committee: Yao-Wen Huang
William L. Kerr
Robert L. Shewfelt
Louise Wicker

Electronic Version Approved:

Maureen Grasso
Dean of the Graduate School
The University of Georgia
May 2012
DEDICATION

To my parents

iv
ACKNOWLEDGEMENTS
I feel a deep sense of gratitude to all the people who contributed towards the
completion of this dissertation.
I want to especially thank my major professor, Dr. Casimir C. Akoh, who not only
supported me in the accomplishment of this research but also encouraged and challenged
me throughout my academic program towards the best realization of my goals.
I express also my deep appreciation to each professor that served on my advisory
committee, Drs. Robert L. Shewfelt, Louise Wicker, William L. Kerr, and Yao-Wen
Huang for their guidance. I would also like to thank USDA National Institute of Food and
Agriculture for their grant to Dr. Akoh in support of this project.
I would like to thank all my lab members for their kind co-operation towards the
completion of my research. A special thanks to Ms. Victoria Wentzel without whose
knowledge and assistance this study would not have been successful.
I would like to thank my family and friends, especially my mother, Dr. Nutan
Pande, my brother, Mr. Gaurav Pande, my husband, Mr. Rajiv Ranjan, and Eddie for
their encouragement and endless love, throughout the duration of my studies.
Last but not least, thanks to the faculty and staff of the Department of Food
Science & Technology, UGA, for being part of my graduate student life and for their
contributions in my personal and professional development.

v
TABLE OF CONTENTS
Page
ACKNOWLEDGEMENTS .................................................................................................v
LIST OF TABLES ........................................................................................................... viii
LIST OF FIGURES .............................................................................................................x
CHAPTER
1 INTRODUCTION .............................................................................................1
2 LITERATURE REVIEW ..................................................................................6
3 ENZYMATIC SYNTHESIS OF TRANS-FREE STRUCTURED
MARGARINE FAT ANALOGUES USING STEARIDONIC ACID
SOYBEAN AND HIGH STEARATE SOYBEAN OILS ...............................57
4 PRODUCTION OF TRANS-FREE MARGARINE WITH STEARIDONIC
ACID SOYBEAN AND HIGH STEARATE SOYBEAN OILS-BASED
STRUCTURED LIPID ....................................................................................91
5 ENZYMATIC SYNTHESIS OF TRANS-FREE STRUCTURED
MARGARINE FAT ANALOGUES WITH HIGH STEARATE SOYBEAN
OIL AND PALM STEARIN AND THEIR CHARACTERIZATION .......125
6 UTILIZATION OF ENZYMATICALLY INTERESTERIFIED
COTTONSEED OIL AND PALM STEARIN-BASED STRUCTURED
LIPID IN THE PRODUCTION OF TRANS-FREE MARGARINE .............158
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7 PRODUCTION OF TRANS-FREE MARGARINE WITH
ENZYMATICALLY INTERESTERIFIED FULLY HYDROGENATED
COTTONSEED OIL AND PALM STEARIN ..............................................198
8 CONCLUSIONS............................................................................................231

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LIST OF TABLES
Page
Table 3.1: Experimental settings of the factors and responses used for optimization by
response surface methodology ...............................................................................79
Table 3.2.1: ANOVA table for incorporation of stearic acid ............................................80
Table 3.2.2: ANOVA table for SDA content .....................................................................81
Table 3.3: Verification of the models using chi-squared test ............................................82
Table 3.4: Total fatty acid (mol%) of the substrates, SLs, and PBs ..................................83
Table 3.5: Positional fatty acid profile (mol%) of the substrates, SLs, and PBs ...............84
Table 3.6: Polymorphic forms of the substrates, SLs, and PBs .........................................85
Table 4.1: Total and positional fatty acid profile of structured lipid and commercial
margarine fat ........................................................................................................116
Table 4.2: Relative percent of triacylglycerol (TAG) molecular species of structured lipid
and commercial margarine fat .............................................................................117
Table 4.3: Tocopherol content and oxidative stability index (OSI) of substrates and large-
scale products .......................................................................................................118
Table 5.1: Experimental settings of the factors and the responses used for optimization by
response surface methodology .............................................................................147
Table 5.2: Total and positional fatty acid composition (g/100 g) of high stearate
cottonseed oil and palm stearin ............................................................................148
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