Grant Information
| Knowledge Area | Subject of Investigation | Field of Science | Percent |
|---|---|---|---|
| 501 - New and Improved Food Processing Technologies | 1119 - Deciduous tree fruits, general/other | 1000 - Biochemistry and biophysics | 30% |
| 501 - New and Improved Food Processing Technologies | 1899 - Oilseed and oil crops, general/other | 1000 - Biochemistry and biophysics | 20% |
| 502 - New and Improved Food Products | 1899 - Oilseed and oil crops, general/other | 1010 - Nutrition and metabolism | 20% |
| 501 - New and Improved Food Processing Technologies | 5010 - Food (not readily associated with specific plant and animal products) | 1000 - Biochemistry and biophysics | 15% |
| 724 - *Healthy Lifestyle | 1899 - Oilseed and oil crops, general/other | 1000 - Biochemistry and biophysics | 15% |
Consumption of too much fat in the diet is linked to heart disease, cancer and obesity. However, an increase in the consumption of polyunsaturated fatty acids (PUFAs) of the n-3 series from fish and n-9 fatty acids (FAs) from certain vegetable oils has been linked to reduced risk of coronary heart disease, which is still the leading cause of death in the Western World. The food industry and researchers are looking for ways to provide the consumer with healthier and functional fats and are focusing attention on the development of structured lipids (SLs), human milk fat analogues, and foods containing n-3 and n-9 FAs.. It is possible to modify natural fats and oils with lipases using the appropriate reaction conditions. Enzymes are specific and selective in their reactions. Enzyme assisted modification of lipids will lead to lipids with specific functionality in foods and positive health outcome.
The main goal is to use enzymes to modify lipids to add value and impact functional and healthful properties. Enzyme-assisted modification of lipids will result in functional and healthful lipids which can be used to formulate food products to benefit infants and adult nutrition. Objectives:
Preparation of FFAs from DHASCO® and ARASCO®. They will be converted to FFAs according to Vázquez and Akoh (2011) with modifications. One hundred and fifty grams of oil will be saponified using a mixture of KOH (34.5 g), distilled water (66 mL), 96% ethanol (396 mL), and butylated hydroxytoluene (0.03 g), by placing the reagents in a 1 L stirred batch reactor with a circulating water bath at 60 °C, for 1 h. The reaction will then be stopped by adding 120 mL of distilled water. The mixture will be acidified by adding 6 M HCl and adjusting to pH 2 to release the FFAs. Hexane used for extraction will be removed using a rotovapor at 40 °C.Enzymatic Synthesis. Lipase-catalyzed acidolysis of substrates will be performed in stirred batch bioreactor. Palm stearin 200-400 g will be mixed with ARA-FFA + DHA-FFA (ARA:DHA ratio will be kept between 1:1 and 2:1) at different molar ratios that range from 2-8 and 10 wt% immobilized Lipozyme TLIM lipase (sn-1,3 specific) and mixed at 200 rpm for 24 h at 60 ºC. Substrates could be tripalmitin, gamma linolenic acid (GLA), high stearidonic acid soybean oil (SDA), Krill oil, anhydrous milk fat, olive oil and canola oil at various combinations can also be used as substrates. Reactions involving TAG and another TAG (enzymatic interesterification) will also be explored to produce healthful SL and infant formula fat analogs. All reactions will be performed in triplicate.Analysis of Products. After enzymatic reaction, the resulting product will be analyzed using silica gel G TLC plates. A mixture of petroleum ether: diethyl ether: acetic acid (70:30:0.5, v/v/v) will be used to separate the TAG (Nagachinta and Akoh, 2012, 2013a,b) from other reaction products. The TAG band will be identified using triolein as standard and visualized under UV light after spraying the plates with 0.2% 2, 7-dichlorofluorescein in methanol. The TAG band will be recovered into test tube for conversion to fatty acid methyl esters (FAME) and positional analysis. TPurification. Short-path distillation (KDL-4 unit, UIC Inc.) will be used to remove FFAs from the SL under the following conditions: holding temperature: 60 °C; feeding rate: ~100 mL/h; heating oil temperature: 185 °C; coolant temperature: 15-20 °C; and vacuum:
Target Audience
Nothing Reported
Changes / Problems
Nothing Reported
Training & Professional Development
The project director and the students participated in professional meetings such as IFT, AOCS, ACS, and World Congress on Oils & Fats (Sydney, Australia) to present results of their findings and to enrich their knowledge by listening to other presentations.
Dissemination Streams
Through publications, seminars, and presentations at professional meetings.
Next Reporting Steps
This is the final report.
Target Audience
Nothing Reported
Changes / Problems
Nothing Reported
Training & Professional Development
The project director and the students attended professional meetings such as IFT, AOCS, Euro Fed Lipids, China AOCS Section, and ISBAB to present results of their findings and to enrich their knowledge by listening to other presentations. One PhD dissertation was produced.
Dissemination Streams
Through publications, seminars, and presentations at professional meetings.
Next Reporting Steps
Will continue our research on enzymatic production of functional and healthful lipids and lipid analysis. <br><br>
<br>What was accomplished under these goals? Menhaden oil was enzymatically modified with caprylic (C8:0) and/or stearic acid (C18:0) to produce structured lipids (SL) with high amounts of polyunsaturated fatty acids (PUFA), low level of saturation, and melting point of 25-35°C. Lipases from Candida antarctica, Lipozyme 435, and Rhizomucor miehei, LipozymeRM IM (Novozymes North America, Inc., Franklinton, NC, USA), were compared for all reactions. The physicochemical properties of the produced SL suggest that some of them may be useful in formulating food products such as margarines and spreads that deliver beneficial n-3 PUFA. Problem with using n-3 PUFA in foods is how to control oxidation and stability. To find solution to this problem, oleogels/organogels were produced using different oleogelators/organogelators. One organogel was formed using a phytosterol blend of β-sitosterol and γ-oryzanol (molar ratios of 2:1, 1:1, and 1:2 at 4, 6, and 8% (w/w) in oil), and another was formed with a blend of sucrose stearate (HLB value: 2) and ascorbyl palmitate (SSAP) (molar ratios of 3:1, 1:1, and 1:3 at 8, 10, and 12% (w/w) in oil). All 1:1 molar ratio blends exhibited β-primepolymorphic form with short spacing peaks at 4.20, 3.97, and 3.71 Å. These oleogels were evaluated as alternatives to shortening in the preparation of yellow cake in terms of batter and cake physicochemical properties. All organogels improved the oxidative stability of the menhaden oil, SL, and some could be substituted for shortening in the yellow cake. The organogels have the potential for use as nutraceuticals or health beneficial low saturated fat alternatives to saturated and/or trans-fats. Encapsulation of the oleogels reduced leaching and further improved oxidative stability of the SLs. Fish oil oxidative stability was also studied in a high fat fish oil-in-water emulsions stabilized with sodium caseinate and phosphatidylcholine (PC) as emulsifiers. In another study, fish oil-loaded elctrosprayed capsules were stabilized with seaweed and commercial natural antioxidants. Mango kernel fat (MKF)-based fats characterized as high levels of 1,3-distearoyl-2-oleoyl-glycerol (StOSt), including physical blend (PB), interesterified blend (IB) and its counterpart non-interesterified blend (Non-IB), were used to manufacture "dark chocolates" by optimizing tempering process. Significant improvements on heat and fat bloom stabilities of PB- and IB-chocolates wereattributedto their optimal triacylglycerol compositions (StOSt=55.7-60.9%, 1-palmitoyl-2-oleoyl-3-stearoylglycerol=21.1-23.8%, and 1,3-dipalmitoyl-2-oleoylglycerol=8.2-11.1%) and plate-like growth at 20 °C. One collaborative review article on improving heat and fat bloom stabilities of dark chocolate using mango kernel fat was published. Another invited collaborative review article on novel antioxidants from seaweed was also published. <br><br><b>Publications</b><br>
Target Audience
Nothing Reported
Changes / Problems
Nothing Reported
Training & Professional Development
The project director and the students attended professional meetings such as IFT, AOCS, Euro Fed Lipids and ISBAB to present results of their findings and to enrich their knowledge by listening to other presentations.
Dissemination Streams
Through publications and seminar presentations at industry and professional meetings.
Next Reporting Steps
Will continue our research on enzymatic production of functional and healthful lipids and lipid analysis. <br><br>
<br>What was accomplished under these goals? Cocoa butter equivalents (CBEs) are fats that can mimic the properties of cocoa butter by having similar triacylglycerols profile to cocoa butter (CB). CBE was produced from illipe butter and palm mid-fraction by enzymatic interesterification. The major TAGs of the interesterified product (IP) were 1-palmitoyl-2-oleoyl-3-stearoylglycerol (POS) at 42.7 ± 1.0%, 1,3-distearoyl-2-oleoylglycerol (SOS) at 29.9 ± 0.3%, and 1,3-dipalmitoyl-2-oleoylglycerol (POP) at 19.1 ± 1.0% which were very similar to CB. Thermal behavior, polymorphism, solid fat content, and crystal microstructure of IP were analyzed and were shown to match CB as well. IP was formulated into dark and white 'chocolates' and their textural, rheological, particle size distribution and fat bloom properties analyzed. The analysis results showed that properties of the 'chocolates' made from IP and CB were similar and therefore suggested that IP is compatible with CB and can be used in chocolate as a CBE. In another experiment, we used mango kernel fat-third stearin (MKF-TS), hard palm-mid fraction (HPMF) and CB to produce hard chocolate fats via physical blending and enzymatic interesterification. The optimal physical blend (PB) produced from 10% HPMF, 55% MKF-TS and 35% CB contained 60.9 g/100g 1,3-distearoyl-2-oleoyl-glycerol, 21.1 g/100g 1-palmitoyl-2-oleoyl-3-stearoylglycerol and 9.5 g/100g 1,3-dipalmitoyl-2-oleoylglycerol. The optimal interesterified blend (IB) enzymatically synthesized from 20% HPMF, 60% MKF-TS and 20% CB shared similar triacylglycerol composition to PB. Both PB and IB showed improved thermostabilities compared to CB in terms of sold fat contents, melting and crystallization behaviors. They also exhibited β polymorphic forms and 50-120 μm featherlike crystals, which were close to those of CB. The melting of IB tended to be sharper than non-interesterified blend (Non-IB) and its crystals were more continuous and uniform, indicating that enzymatic interesterification is a potential technique for manufacturing hard chocolate fats. We also used a rice bran oil solid fat fraction (RBOSF) to produce cocoa butter alternatives via interesterification reaction catalyzed by immobilized lipase (Lipozyme® RM IM) in hexane. Effects of reaction time (6, 12, and 18 h), temperature (55, 60, and 65ºC), mole ratios of 3 substrates [RBOSF:palm olein:C18:0 donors (1:1:2, 1:2:3, and 1:2:6)] were determined. The substrate system put in 3 ml of hexane and 10% of lipase added. Two sources of C18:0 donors, stearic acid (SAd) and ethyl stearate (ESd) were used. Optimal reaction was 1:2:6 mole ratio of the substrate (RBOSF:PO:SAd), at 65ºC for 12 h. DSC curves showed the melting point of CB at 20.94ºC, while those of the SL were 14.15 and 40.35ºC, respectively. The melting completion temperature (Tmc) of CB was 25.5°C while that of SL was 43.9°C, respectively. Healthful structured lipids (SLs) were produced using menhaden oil and capric acid or ethyl caprate as substrate. Enzymatic reactions conditions were optimized using the Taguchi method L9 orthogonal array with three substrate molar ratio levels of capric acid or ethyl caprate to menhaden oil (1:1, 2:1, and 3:1), three enzyme load levels (5, 10, and 15% [w/w]), three temperature levels (40, 50, and 60?C), and three reaction times (12, 24, 36 h). Reactions with ethyl caprate incorporated significantly more C10:0, at 30.76±1.15 and 28.63±2.37 mol% versus 19.50±1.06 and 9.81±1.51 mol%, respectively, for both Lipozyme® 435 and Lipozyme® RM IM, respectively. Reactions with ethyl caprate as substrate and Lipozyme® 435 as biocatalyst produced more of the desired medium-long-medium (MLM)-type TAGs with polyunsaturated fatty acids (PUFAs) at sn-2 and C10:0 at sn-1,3 positions. One review article on mango kernel fat fractions as potential healthy food ingredient and one essay based on the IFT Babcock-Hart Award received in 2018 for contributions that resulted in improved public health through nutrition or more nutritious food were published. <br><br><b>Publications</b><br>
Target Audience
Nothing Reported
Changes / Problems
Nothing Reported
Training & Professional Development
The project director and the students attended professional meetings such as IFT, AOCS, EuroFed LIpids and ISBAB to present results of their findings and to enrich their knowledge by listening to other presentations.
Dissemination Streams
Through publications and seminar presentations at industry and professional meetings.
Next Reporting Steps
Will continue our research on enzymatic production of functional and healthful lipids. <br><br>
<br>What was accomplished under these goals? A glucose-cysteine Maillard reaction product (MRP) was produced and its antioxidant effects on lipid oxidation were determined for a structured-lipid enriched with polyunsaturated fatty acids in a complex emulsion. Trolox equivalent antioxidant capacities (TEAC) were determined for MRP heating intervals of 2, 4, and 6 h and were compared to α-tocopherol (TOC), MRP with TOC (TOC-MRP), and TOC with ascorbyl palmitate (TOC-AP). Emulsions were produced with total antioxidant additions of 0.02% of the oil, and lipid oxidation was monitored by peroxide and p-anisidine values over 56 d. Positive correlations between browning and heating time as well as TEAC were observed. Total TEAC values for the MRP at 6 h, TOC, TOC with the MRP at 6 h, and TOC-AP were 2.51, 3.87, 2.68, and 2.76 mg trolox eq/g, respectively. Oxidation results indicated a possible antioxidant effect for the MRP at 6 h on secondary oxidation for days 14 to 28. These results suggest that the MRP at 6 h could be useful in inhibiting secondary oxidation in complex emulsions. In another study we demonstrated that ultrasound along with supercooling can induce a change in the physical properties of interesterified (IE) fat samples including microstructure, isothermal solid fat content (SFC), and rheological properties. Although the application of high intensity ultrasound (HIU) did change the physical properties of both IE with 20 and 30% palmitic acid, the effect was more pronounced on the IE with 30% palmitic acid. This could be due to differences in TAG composition between physical blend (PB) and IE samples. Palmitic acid was mostly present in the OOP + OPP TAG in IE samples while PB samples were composed mainly of PPP. These TAG species are responsible for driving the crystallization of the samples. The higher concentration of these TAG in the IE 30% palmitic acid sample may have corresponded to more nuclei and thus higher degree of crystallization, which may have affected the physical properties of these samples upon sonication. Results from this research suggest that HIU effectiveness is driven by two main factors: (a) the generation of supercooling, and (b) the presence of sufficient saturated fats. That is, HIU is not effective at inducing crystallization in samples with low content of palmitic acid. It is possible that the lack of effect under these conditions is due to physical properties of the material such as its viscosity, which will affect the formation of cavities during sonication. However, with slightly higher content of palmitic acid (30%), sonication and processing conditions can be tailored to obtain various physical properties. By changing the HIU conditions, the extent of change may be modified and this processing technique can be extended to the healthier IE samples for use as trans-free fat alternatives. We performed similar experiment using stearic acid (S) in place of palmitic acid.Sonication promoted crystallization of low melting TAGs and the incorporation of SSS into the crystalline network. The IE samples with stearic acid at the sn-2 position have superior crystallization properties including SFC and rheology than the IE with palmitic acid at the sn-2 position. Although HIU was not as effective at inducing crystallization in the IE C16:0 20% samples due to the lower amount of saturated fats in the system, HIU induced crystallization in both the IE C18:0 20 and 30% samples. This could have been due to the higher melting point of the stearic containing samples compared to the palmitic ones. The induction of superior crystallization properties in these samples upon sonication can make them great candidates as ingredients for trans-fat free applications. Structured lipids were designed for use in an edible film application and their properties compared. Blends [60:40, 70:30, and 80:20 (w/w)] of coconut oil (CO) and high oleic sunflower oil (HOSO) were interesterified using immobilized enzyme, Lipozyme® TL IM. The enzyme was successful in increasing oleic acid at the sn-2 position of the TAGs. The structured lipids were helpful in reducing the opacity of CO in a film use and this is advantageous. The interesterified product (IP) 60:40 provided the strongest tensile strength (TS) although it had the weakest elongation break (EB). The IP 60:40 contained the most oleic acid (33.86 ± 1.55%) at the sn-2 position and also resulted in a fairly translucent product. The IP 60:40 was used to continue further research on the use of SLs in edible films to prepare sports nutrition products in our laboratory. Two review articles were published on the structured lipids for food and nutraceutical application and on biotechnological and novel approaches for designing structured lipids for infant nutrition. <br><br><b>Publications</b><br>
Target Audience
Nothing Reported
Changes / Problems
Nothing Reported
Training & Professional Development
Students and the project director attended professional meetings such as AOCs and IFt to present the results and enrich their knowledge by listening to other presentations.
Dissemination Streams
Through publications, seminars, industrial presentations, and professional meeting presentations
Next Reporting Steps
To continue our research on production of healthful and functional lipids using enzymes and use them in food applications. <br><br>
<br>What was accomplished under these goals? We compared encapsulation of stearidonic acid soybean oil (SDASO) by complex coacervation in the classical gelatin (GE)-gum arabic (GA) system with that of a Maillard reaction product (MRP). A portion of the control (microcapsules based on the GE-GA system) was cross-linked with transglutaminase (TG) after encapsulation.Based on the amount of oil released from microcapsules during heat treatment (85 C for 30 min) in yogurt milk base, MR-modified microcapsules displayed the highest thermal stability. Furthermore, yogurts formulated with MR-modified microcapsules had the best oxidative stability during 14 days of storage at 4 C demonstrating that the antioxidant components of MR-modified microcapsules had good carry-through properties. High sn-2 docosahexaenoic and arachidonic acid oils (DHAOm and ARAOm, respectively) were produced independently via enzymatic interesterification of DHA-rich and ARA-rich single cell oils (DHASCO and ARASCO, respectively) using a mix of immobilized lipases, Lipozyme® TL IM and Novozym® 435 (weight ratio 1:1) as the biocatalyst system. Final results showed that DHAOm contained 46.53 mol% of total DHA (49.70 % at the sn-2 position), while ARAOm contained 47.25 mol% of total ARA (36.08 % at the sn-2 position). This represents a significant increment in the amount of DHA and ARA at the sn-2 position when compared to DHASCO (47.8 mol%; 30.30 % at the sn-2) and ARASCO (47.79 mol%; 28.50 % at the sn-2), respectively. These products have potential as additions to infant formulas where DHA and ARA supplementation is required. A structured lipid (SL) with a high amount of sn-2 palmitic acid was synthesized from anhydrous milkfat and was then enriched with docosahexaenoic (DHA) and arachidonic (ARA) acids using an immobilized lipase. This SL might be totally or partially used in commercial fat blends for infant formula. <br><br><b>Publications</b><br>
Target Audience
Nothing Reported
Changes / Problems
Nothing Reported
Training & Professional Development
Nothing Reported
Dissemination Streams
Nothing Reported
Next Reporting Steps
Nothing Reported
<br><br>
<br>What was accomplished under these goals? Nothing to report. Project is new. <br><br><b>Publications</b><br>