Effect of D-allulose on rheological properties of chicken breast sausage

M. Hadipernata,∗,† M. Ogawa,‡,1 and S. Hayakawa‡

∗Department of Applied Bioresource Science, The United Graduate School of Agricultural Sciences, EhimeUniversity (Affiliated University: Kagawa University), 2393 Ikenobe, Miki-cho, Kita-gun, Kagawa 761–0795,Japan; †Agency for Agricultural Research and Development, Indonesia Ministry of Agriculture, Jl. TentaraPelajar 12, Bogor 16111, Indonesia; and ‡Department of Applied Biological Science, Faculty of Agriculture,

Kagawa University, 2393 Ikenobe, Miki-cho, Kita-gun, Kagawa 761–0795, Japan

ABSTRACT d-Allulose (Alu), a rare sugar, was ap-plied to chicken breast sausage as a sucrose (Suc) sub-stitute. The ratio (w/w) of Alu to Suc in sugar thatwas added to the sausage batter was 0/1 (A0S1), 3/7(A3S7), 7/3 (A7S3), and 1/0 (A1S0). The total amountof Suc used was 2.5% of the weight of minced chickenbreast meat. Substituting Suc with Alu did not af-fect water content, cooking loss, breaking stress, break-ing strain, and modulus of elasticity of chicken breastsausage, but a 100% substitution with Alu caused a10% decrease in viscosity and a 31% decrease in ex-

pressible water. A significant difference appeared in therheological properties of elasticity, viscosity, and water-holding capacity of chicken breast sausage frozen-stored(−20◦C) for 90 d. Particularly, the modulus of elasticityfor A1S0 chicken breast sausage was 19% higher thanthat of the control A0S1 chicken breast sausage, sug-gesting that Alu appreciably reduced the deteriorationin elasticity that is caused by long-term frozen storageof sausage. The quality improvement of frozen-storedchicken breast sausage demonstrates the feasibility andbenefits of the application of Alu to frozen foods.

Key words: chicken breast sausage, d-allulose, expressible water, frozen food, rare sugar2016 Poultry Science 95:2120–2128

http://dx.doi.org/10.3382/ps/pew143

INTRODUCTION

Chicken breast sausage is widely consumed in manycountries. Especially in Islamic countries, it is a mainfood commodity. In Indonesia, the country with thelargest Islamic population in the world, chicken breastsausage occupies 43% of sausage market shares and thesausage market has grown 32.7% in five years, since2007 (Wulandari et al., 2014). Chicken breast sausage isthought to be a healthful food because of its low fat con-tent and high protein content (Ali et al., 2011). On theother hand, sausage in general contains large amountsof ingredients that are linked to lifestyle diseases, suchas salt and sugar. Thus, excessive consumption of foodswith high sugar/fat content can increase the risk of de-veloping lifestyle-related diseases such as hypertensionand type 2 diabetes (Johnson et al., 2009; Dominguezet al., 2015). Type 2 diabetes, in particular, has be-come a global problem. According to the InternationalDiabetes Federation (IDF), 382 million people world-wide have diabetes, and more than 85% of the cases aretype 2 diabetes. In 2013, 5.1 million people died of dia-betes and cost $548 billion (USD) in health care spend-

C© 2016 Poultry Science Association Inc.Received October 29, 2015.Accepted March 11, 2016.1Corresponding author: ogawa@ag.kagawa-u.ac.jp

ing (IDF, 2013). World Health Organization (WHO)guidelines recommend adults and children reduce theirdaily intake of free sugars to less than 10% of their to-tal energy intake (WHO, 2015). Thus, consumers payattention to foods with reduced sugar contents (Bakerand Friel, 2014).

d-Allulose (Alu), also known as d-Psicose, is the C-3epimer of d-fructose (Fru). It is a rare sugar that existsin small amounts in nature (Granstrom, et al., 2004).A method of mass production of Alu was developed bythe Izumori research group (Takeshita, et al., 2000). Ac-cording to Han (2015), Alu can be sold at less than $15per pound in the United States. Alu has a caloric valueof 0.39 kcal, which is roughly equivalent to 10% of thecaloric value of sucrose (Suc) and suppresses the eleva-tion of after-meal blood glucose levels (Matsuo et al.,2002; Hayashi et al., 2010; Hossain et al., 2015). Aluwas approved and generally recognized as safe (GRAS)by the U.S. Food and Drug Administration (FDA) inJune 2014 (GRAS Notice No. GRN 498) (Hossain et al.,2015), and it can be used as an ingredient in foods anddietary supplements (Mu et al., 2012).

Alu also has demonstrated promising applications foruse in the food industry. It has been shown to improvethe food properties of some processed foods and theirmaterials. The application of Alu as a Suc substituteto meringue-based confectionery resulted in a crunchiertexture and enhanced the antioxidant activity of baked

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meringue (O’Charoen et al., 2014). Custard pudding,utilizing Alu as a Suc substitute, demonstrated higherbreaking strength and higher viscoelasticity than thecontrol with Suc as well as with Fru (Sun et al., 2006).Furthermore, Alu demonstrated better foaming prop-erties of egg white protein compared with Suc and Fru(Sun et al., 2008). However, there is little research onthe effects of Alu on the processing characteristics ofmeat. In this study, we determined the effects of Alu asa Suc substitute on the rheological properties of chickenbreast sausage. We also investigated the effects of Aluon frozen-storage of chicken breast sausage. This is thefirst report on the application of Alu to sausage.

MATERIALS AND METHODS

Materials

Chicken breast meat, salt, and Suc were purchasedfrom a local market. Alu (PubChem CID: 441036) wasobtained from the Rare Sugar Research Center, Ka-gawa University, Japan. Sodium polyphosphate waspurchased from Wako Pure Chemical Industries, Tokyo,Japan. All other reagents used were of analytical grade.

Sausage Preparation

Chicken breast halves (3 kg) were diced and groundthrough a grinder (KitchenAid, Model KSM5; StJoseph, Michigan) with 4 mm plate. Cold water(40 mL), sodium chloride (3.6 g), sodium polyphos-phate (1 g), and sugar (5 g) were added to 200 g ofthe minced meat in a chilled bowl and mixed by handfor 3 min. The 4 chicken breast sausage treatments forsugar were 0:100 (A0S1), 30:70 (A3S7), 70:30 (A7S3),and 100:0 Alu:Suc (A1S0). The mixture was homoge-nized for 30 s with a food processor (TK-551; TescomCo., Ltd., Tokyo, Japan). Then, the resulting meat bat-ter was left for 10 min in a bowl on ice, and mixed againfor 30 s with the food processor. The batter was stuffedinto a Krehalon R© PVDC casing (flat width, 4.0 cm;Kureha Co., Tokyo, Japan) with a handmade sausagestuffer. Both ends of the stuffed sausage were closedwith kite strings. The stuffed meat batter was heatedin a water bath at 70 ± 1◦C for 30 min. The diameterof heated sausages was 25 mm. Immediately after boil-ing, the chicken breast sausage gels formed were cooleddown by sitting at room temperature for 3 h. After re-moval of the casing, the chicken breast sausages wereanalyzed or frozen at −20◦C in a still freezer (Biomed-ical Freezer, MDF-436; Sanyo, Japan). The chickenbreast sausages were packaged in a polypropylene plas-tic box and stored frozen at −20◦C for 30 or 90 d beforethawing in a refrigerator (4◦C) for 16 h before analysis.A sausage batter without sugar was prepared for DSCanalysis only. The sausage batter without sugar derivedfrom 50 g of minced meat, and the percentage of ingre-

dients and treatment stages were the same as makingsausage batter with sugar.

Determination of Water Content

One gram of sausage pieces was precisely weighedin an aluminum pan. The pieces were dehydrated in adrying oven set at 105 ± 1◦C until the sample weightreached a constant value. The dehydrate was cooleddown in a desiccator in which silica gel had been placedand then weighed with a precision balance. The watercontent of the sausage was calculated using the follow-ing equation:

Water content (%) = (Wi − Wf )/Wi × 100

where Wi is the initial weight of sausage pieces (beforedrying) and Wf is the final weight of the dehydrate.

Cooking Loss

Cooking loss was calculated from the weight differ-ence between uncooked and cooked samples (Jin et al.,2007). Cooking loss of sausage was calculated using thefollowing equation:

Cooking loss (%) = (Ci − Cf ) /Ci × 100

where Ci is the initial weight of uncooked sausage andCf is the weight of cooked sausage.

Determination of Expressible Water

The expressible water of sausages was measured bythe method developed by Uresti et al. (2003). A 6 mmsausage slice was precisely weighed and placed between2 layers of filter paper. The slice was sandwiched be-tween the 2 filter papers placed at the bottom of a50 mL centrifuge tube and centrifuged at 1,000 × g for15 min at 15◦C. Immediately after centrifugation, thecompressed sausage slice was weighed. Expressible wa-ter of sausage was calculated using the following equa-tion:

Expressible water (%) = (Wi − Wc) /Wi × 100

where Wi is the initial weight of the sausage slice andWc is the weight of the compressed slice. All measure-ments were performed on 3 replicate experiments, eachexperiment consisting of 3 measured samples. Resultsare the mean of 9 samples for each treatment.

Rheological Properties

The rheological properties of sausage were assessedby 2 methods — a breaking test and a creep test— using a Rheoner II creep meter (RE 2–3305;Yamaden Co., Tokyo, Japan). The breaking test was

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conducted using the creep meter equipped with a cylin-drical plunger (diameter 5 mm). A piece of sausage wascut at intervals of 20 mm with a cutter knife to get cylin-drical sausage gels (diameter 25 mm, height 20 mm). Acut gel sample was put on the stage of the rheometerand its breaking stress and breaking strain were mea-sured at a penetration speed of 1.0 mm/s. The breakingstress and strain were represented as the stress (N/m2)and strain (%) at the top of the first peak of the stressvs. strain curve.

The creep test was conducted using the creep meterequipped with a 4 cm diameter plate plunger. A cylin-drical sausage gel sample with the same size (diameter25 mm, height 20 mm) was put on the stage of thecreep meter and its creep measurement was carried outunder uniaxial compression at 10 mm/s over a periodof 2 min. According to Sun and Hayakawa (2002), thecreep curve obtained was analyzed by a 6-element me-chanical model as described by equation 1 based on therelationship between stress and strain:

ε(t) =s

E0+ Σ2

i=1s

Ei

(1 − e

(− t

τ i

))+

(s

ηN

)t (1)

where ε(t) = strain (dimensionless), S = stress (N/m2),E0 = elastic modulus of a Hookean body (instantaneousmodulus; N/m2), Ei = elastic modulus of a Voigt body(N/m2), τ i = retardation time (s), ηN = Newtonianviscosity (Pa·s), and t = time (s).

Viscoelastic parameters were calculated from thecreep compliance data according to Eqs. 1 to 3 (Peleg,1980):

J(t) =ε(t)s

(2)

J (t) = J0 + Σ2i=1Ji

(1 − e

(− t

τ 1

))+ t/ηN (3)

where J(t) = creep compliance (m2/N), J0 = instan-taneous compliance (m2/N), Ji = retarded compliance(m2/N), and t = time (s).

Determination of Alu and Suc

Concentrations of Alu and Suc in sausage were deter-mined by the high-performance liquid chromatography(HPLC) method of Oshima et al. (2006) with slightmodifications. Ten grams of sausage batter or the sameweight of sausage pieces (2 × 2 × 2 mm3) were homoge-nized using a mini-chopping machine (Iwatani Int. Co.,Tokyo, Japan). An appropriate amount of homogenatewas precisely weighed in a beaker, to which 40 mL ofHPLC grade water was added. Sugar contained in thesausage was extracted from the suspension using ul-trasonic treatment (Sharp UT-204, Sharp Co., Tokyo,Japan) at 20◦C for 10 min. The extracted sugar wascentrifuged at 3,500 × g for 5 min and filtered through

4-layer cheese-cloth. The resultant pellet was re-extracted with 40 mL of HPLC grade water and thenthe combined supernatants were diluted to 100 mL withHPLC grade water. The extracted sugar solution (500μL) was taken and mixed with the 2 ion exchange resinsof strongly acidic cation-exchange resin DIAIONTM

SKIB (Mitsubishi Chemical Co., Tokyo, Japan) andstrong basic anion-exchange resin Amberlite IRA411(Organo Co., Ltd., Tokyo, Japan) and let stand for30 min to remove charged substances. The sugar so-lution was filtered through a disposable membrane fil-ter unit with pore size 0.2 μm. The pretreated sugarsolution was subjected to sugar analysis with a Shi-madzu HPLC system equipped with a RID-10A refrac-tive index detector (Shimadzu Co., Kyoto, Japan) anda Hitachi Gel pack GL-C611 column (Hitachi Chemi-cal Co., Ltd., Tokyo, Japan). Two hundred microlitersof sugar solution were injected into the GL-C611 col-umn and eluted with 0.1 mM NaOH at the flow rate of1.0 mL/min (column temperature, 60◦C). The concen-trations of Suc and Alu were determined from calibra-tion curves of their standards.

Differential Scanning Calorimetry

Thermal denaturation of proteins in the sausage bat-ter was determined by differential scanning calorimetry(DSC). A Setaram Micro-DSC VII Commissioning/Utilizations (SETARAM Instrumentation, Caluire,France) was employed for DSC analysis. One hundredmilligrams of sausage batter and 100 milligrams ofAl2O3 were applied to a sample pan and a referencepan, respectively. The heating rate was 1.2◦C/s in thetemperature range of 20 to 90◦C. The calorimetric datawere analyzed using thermal analysis software providedwith the DSC instrument.

SDS-Polyacrylamide Gel Electrophoresis

The protein profile of the sausage was analyzedby the Sodium Dodecyl Sulfate Polyacrylamide GelElectrophoresis (SDS-PAGE) method. One gram ofcooked sausage sample was minced and homogenizedusing a Nissei AM-8 homogenizer (Nihonseiki KaishaLtd., Tokyo, Japan) at 6,500 rpm for one min in 10 mLdissociation solution (0.6 M NaCl, 8.0 M urea, 1.25%SDS, and 20 mM sodium phosphate buffer, pH 8.0).The homogenate was then shaken with a SR-1 shaker(AS ONE Co., Osaka, Japan) at 4◦C for 12 h beforeit was centrifuged at 4,500 × g for 20 min. The su-pernatant (40 μL) was mixed with 10 μL of SDS sam-ple buffer (100 mM Tris-HCl buffer, pH 6.7, containing5.0% SDS and 0.004% bromophenol blue with 2.0% β-mercaptoethanol) and the mixture was heated at 90◦Cfor 5 min. The SDS sample solution thus prepared(10 μL) was put into wells of SDS-PAGE gel with 10%acrylamide separating gel. SDS-PAGE was carried out

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according to the method of Laemmli (1970). The gelwas stained with 0.025% coomassie brilliant blue.

Statistical Analysis

This experiment was set up using a completely ran-domized design (Oehlert, 2010). Four different chickenbreast sausage treatments were tested with 3 replicatesper experiment for each treatment. Each experimentconsisted of 3 sample measurements. Statistical analysiswas performed using the software SPSS 15.0 (SPSS Inc.,Chicago, IL) by one-way analysis of variance (ANOVA)and the statistical difference of mean was determinedby the Duncan multiple-range test with a significantlevel of 95%.

RESULTS AND DISCUSSION

Chemical Properties of Sausage

Table 1 shows water content, cooking loss, sugarcontent, and expressible water of the chicken breastsausage. Water content did not differ among A0S1,A3S7, A7S3, and A1S0. This shows that the sub-stitution of Suc to Alu does not affect moistureretained in boiled sausage. Also, cooking loss ofsausages did not differ significantly among all treat-ments. The cooking loss values of A0S1, A3S7,A7S3, and A1S0 were 10.67, 10.32, 10.45, and 9.93%,respectively.

Sugar content in chicken breast sausage was 24 to33% lower than that of the sausage batters. The cook-ing loss of Alu was 29 to 33%, which was higher thanthat of Suc (24 to 29%). The higher loss in Alu maybe due to the higher degree of Maillard reaction andthe faster isomerization reaction of Alu, because Aluis a reducing sugar while Suc is a non-reducing sugar.Oshima et al. (2014) reported that in several foodsAlu content decreased during cooking processing. Aluloss was 10.8% for sponge cake and 7.7% for bakedmeringue. The large difference in Alu loss is proba-

Figure 1. Sodium dodecyl sulfate-PAGE patterns of chicken breastsausage containing d-allulose (Alu).

bly due to differences in cooking methods (baking andboiling). Unlike baking, boiling will cause sugar leak-age from sausage batter to boiling water (Bach et al.,2013).

Sun et al. (2006) reported that Alu facilitated pro-tein cross-linkage in heat-induced gel derived fromegg white protein. To examine if a similar phe-nomenon occurs in chicken sausage, SDS-PAGE anal-ysis of chicken breast sausage protein was carriedout. As shown in Figure 1, the protein-band patternwas similar among A0S1, A3S7, A7S3, and A1S0. Inall the samples tested, a slight band was observedat the top part in the stacking gel of SDS-PAGE,suggesting that high molecular weight protein com-plexes were formed via covalent bonding. The bandintensity in the stacking gel was almost the same

Table 1. Water content, cooking loss, sugar content, and expressible water of sausages.

Treatment1

Item (%) A0S1 A3S7 A7S3 A1S0

Water content 75.51 ± 0.71 75.85 ± 0.79 75.81 ± 0.64 75.99 ± 0.37Cooking loss 10.67 ± 0.37 10.32 ± 0.62 10.45 ± 0.19 9.93 ± 0.54Suc content before cooking 2.32 ± 0.26 1.61 ± 0.11 0.66 ± 0.05 N.D.Suc content after cooking 1.71 ± 0.15 1.14 ± 0.09 0.50 ± 0.01 N.D.Suc loss 26.29 29.19 24.24 N.A.Alu content before cooking N.D. 0.62 ± 0.22 1.56 ± 0.03 2.22 ± 0.06Alu content after cooking N.D. 0.44 ± 0.03 1.05 ± 0.06 1.53 ± 0.11Alu loss N.A. 29.03 32.69 31.08Expressible water 13.64 ± 0.74a 11.68 ± 0.18a,b 10.22 ± 0.18a,b 9.44 ± 0.14b

N.D., not detected; N.A., not available; Data are presented as mean ± SD (n = 9) and values with different superscriptletters (a and b) in a line show the significant difference (P < 0.05).

A0S1 = 0% Alu and 100% Suc; A3S7 = 30% Alu and 70% Suc; A7S3 = 70% Alu and 30% Suc; A1S0 = 100% Aluand 0% Suc.

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Table 2. Rheological properties of sausages.

Treatment1

Item A0S1 A3S7 A7S3 A1S0

Breaking stress (105 N/m2) 2.26 ± 0.08 2.25 ± 0.24 2.23 ± 0.23 2.22 ± 0.29Breaking strain (%) 37.49 ± 3.92 36.09 ± 2.40 37.59 ± 2.15 38.74 ± 1.96Modulus of elasticity E0 (105 N/m2) 1.66 ± 0.04 1.64 ± 0.08 1.62 ± 0.03 1.62 ± 0.04Coefficient of viscosity ηN (107 Pa s) 5.01 ± 0.53a 4.96 ± 0.04a 4.91 ± 0.20a 4.45 ± 0.07b

Data are presented as mean ± SD (n = 9) and values with different superscript letters (a and b) ina line show the significant difference (P < 0.05).

1A0S1 = 0% Alu and 100% Suc; A3S7 = 30% Alu and 70% Suc; A7S3 = 70% Alu and 30% Suc;A1S0 = 100% Alu and 0% Suc.

between A0S1 and A1S0. Thus, Alu seems not to fa-cilitate protein cross-linkage in sausage.

Rheological Properties of Sausage

A comparison of the results is shown in Table 2. Sub-stituting Suc with Alu did not affect breaking stress,breaking strain, or modulus of elasticity of chickenbreast sausage. However, unlike chicken breast sausage,the breaking strength of custard pudding gel consist-ing of egg, milk, and sugar was affected by substi-tuting Suc with Alu. Sun et al. (2006) showed thatcustard pudding gel containing Alu has higher break-ing strength values than counterparts containing Suc.The discrepancy in the results between custard pud-ding and chicken breast sausage is due to the differencein gel forming protein (egg white protein and myofib-rillar protein) and the difference in sugar content inthe food systems (chicken breast sausage contains onlyone-sixth sugar content of custard pudding). On theother hand, substitution with Alu affected the coeffi-cient of viscosity of chicken breast sausage. A 100%substitution with Alu resulted in an 11% decrease inviscosity.

A comparison of the expressible water of chickenbreast sausage is shown in Table 1. The applicationof Alu did make a remarkable difference in express-ible water. A higher substitution rate by Alu led toa lower expressible water. Expressible water of A1S0sausage was only 69% that of A0S1 sausage. Althoughthe decrease in expressible water seems not to be cor-related closely with the decrease in viscosity of sausagegel, an indirect relationship might exist between the2 parameters.

Water-holding capacity of gel is inversely associatedwith the percentage of water expressed by centrifuga-tion (Das et al., 2008). The decrease in expressible wa-ter of chicken breast sausage by substituting Suc withAlu implies that Alu enhances the water-holding ca-pacity of sausage, suggesting that Alu helps chickenbreast sausage proteins to tightly interact with watermolecules. The low expressible water of Alu-chickenbreast sausage might be related to its lower viscosity.

Figure 2. Differential scanning calorimetry (DSC) thermograms ofsausage batters.

Formation of heat-induced protein gel is initiated byheat-denaturation of protein. The effect of Alu on ther-mal denaturation of proteins in sausage batter was in-vestigated by DSC. Figure 2 shows DSC thermogramsof sausage batters. Three major endothermic peaks at53, 62, and 69◦C were observed in all the samples ex-amined. According to Ali et al. (2015), chicken breastmeat shows 3 endothermic transitions at peak temper-atures — 53 to 55, 62 to 64, and 72 to 76◦C — cor-responding to the denaturation of myosin, connectivetissue (together with sarcoplasmic proteins), and actin,respectively. Li et al. (2015) reported that DSC peaks ofchicken breast meat batter containing salt are assignedto myosin (54.52◦C), sarcoplasmic proteins/collagen(62.36◦C), and actin (73.65◦C). Thus, the peak at 53◦Cand the peaks at 62 and 69◦C are presumed to originatein the denaturation of myofibril protein and the de-naturation of sarcoplasmic protein/collagen and actin,respectively. Myofibrillar protein (especially myosin) isa key protein for gelation. That an endothermic peakat 53◦C appeared in all the samples (A0S1, A3S7,A7S3, A1S0, and no sugar) indicates that Alu and Suchave no impact on the denaturation temperature ofthe myofibrillar protein that induces gel formation ofsausage.

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The denaturation enthalpy (ΔH) of 53◦C-peak, de-rived from myofibrillar protein, was calculated for allthe samples. The ΔH values of A0S1, A3S7, A7S3,A1S0, and sausage batter without sugar were 0.1958,0.1316, 0.0894, 0.0818, and 0.0395 J/g, respectively.Thus, substituting Suc with Alu results in a decreasein ΔH. The thermal denaturation of many proteins isaccompanied by aggregation (Johnson, 2013). The dif-ference in ΔH value between A0S1 and A1S0 might bedue to the difference in aggregation rate and/or aggre-gation manner of myofibrillar protein. Low ΔH valuesimply extensive unfolding of protein, which promotes itsthermal gelation process (Kato, et al., 1990; Planckenet al., 2007). Thus, substituting Suc with Alu may causemyofibrillar protein to undergo more extensive unfold-ing, although it did not make a significant difference inthe gel strength.

Rheological Properties of Frozen-storedSausage

Frozen storage is an important preservation methodof foods, especially in terms of bacterial growth. How-ever, frozen storage causes deterioration in food texture(Ramadhan et al., 2012; Benjakul et al., 2005). Rhe-ological properties of chicken breast sausage stored at−20◦C were evaluated to investigate the effect of Alu onthe deterioration prevention of frozen sausage. The re-sults of breaking test, creep test, and the measurementof expressible water are summarized in Figure 3. In all4 samples tested, breaking stress decreased with thestorage period (Figure 3a). Breaking stress of sausagefrozen-stored for 90 d was slightly higher in A1S0 thanin A0S1. In contrast to breaking stress, breaking strainincreased with the storage period (Figure 3b). In the 3storage periods of zero, 30, and 90 d, breaking strainof A1S0 was slightly higher than that of A0S1. Modu-lus of elasticity E0 decreased with the storage period(Figure 3c). A significant difference appeared in E0values of sausages stored for 90 d. A3S7, A7S3, andA1S0 had 7, 16, and 19%, respectively, higher E0 val-ues than A0S1, suggesting that Alu appreciably sup-presses the deterioration in elasticity that is caused bylong-term frozen storage of sausage. The coefficient ofviscosity ηN also decreased with the storage period (Fig-ure 3d). The decreasing rate of viscosity during frozenstorage was lower in A1S0 than in A0S1, A3S7, andA7S3. A1S0 sausage produced only 17% decrement inηN by 90 d frozen storage, while A0S1 sausage hada much higher ηN decrement of 32%. Thus, substitu-tion of Suc with Alu was shown to greatly suppressdeterioration in viscosity. Expressible water increasedwith the storage period (Figure 3e). This clearly showsthat frozen storage of chicken breast sausage induces adecrease in water-holding capacity. The water-holdingcapacity of frozen-stored chicken breast sausage was

significantly higher in A1S0 than in A0S1 and A3S7.Thus, a 100% substitution of Suc with Alu helps toprevent the water-holding capacity of sausage from de-creasing by frozen storage. This high water-holding ca-pacity of A1S0-chicken breast sausage may have pro-duced a deterioration suppression in elasticity andviscosity.

Creep-compliance curves of A0S1 and A1S0 sausagesare shown in Figure 4. An outstanding difference wasseen in the creep recovery curve (time, 60 to 120 s) thatrepresents elastic recovery. Fresh A0S1 sausage (zero dstorage) showed a similar creep recovery curve to freshA1S0 sausage, while creep recovery curves of frozen-stored sausages differed between the 2 sausages. Creeprecovery curves of A0S1 sausage significantly shiftedupward with the storage period. On the other hand, theupward shift of A1S0 sausage was much less than thatof A0S1 sausage. The data indicate that Alu markedlysuppresses the loss of elastic recovery induced by frozenstorage. The suppressive effect of elastic recovery lossis probably due to the high water-holding capacity thatAlu confers to sausage. A similar effect is reported ina disaccharide trehalose. Ramadhant et al. (2012) re-ported that duck surimi-like material containing tre-halose was able to retain the protein solubility, gelbreaking force, and deformation after 4 mo of frozenstorage due to the high water-holding capacity. Thedrip loss that takes place during thawing of frozen meatand meat products and concomitant texture deterio-ration are undesirable for frozen foods. Consequently,Alu is useful in frozen food application of sausagebecause Alu can suppress texture deterioration ofsausage.

In conclusion, the substitution of Suc with Alu, asweetener with no calories, believed to have health ben-efits, did not affect breaking stress, breaking strain,and modulus of elasticity of fresh chicken breastsausage, but 100% substitution with Alu caused a10% decrease in viscosity and 31% decrease in ex-pressible water. The deteriorations in the physicalproperties (elasticity, viscosity, water-holding capacity,and elastic recovery) of chicken breast sausage thatare caused by frozen storage were greatly suppressedby the substitution of Suc with Alu. Thus, Alu wasshown to confer resistance to freezing-related damagein sausage. In particular, the quality improvement offrozen-stored chicken breast sausage demonstrates thefeasibility and benefit of application of Alu to frozenfoods.

ACKNOWLEDGMENTS

This work was partially supported by the City AreaProgram, the Ministry of Education, Culture, Sports,Science & Technology, Japan. The authors also thankPeter Lutes, associate professor of Faculty of Agricul-ture, Kagawa Univ., for language editing.

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Figure 3. Changes in rheological properties and expressible water of chicken breast sausages by frozen storage. (a) Breaking stress, (b)breaking strain, (c) modulus of elasticity, (d) coefficient of viscosity, and (e) expressible water. Different letters indicate significant differencebetween samples in the same storage period (P < 0.05).

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Figure 4. Creep-compliance curves of sausages frozen-stored at−20◦C. (a) A0S1 and (b) A1S0.

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