• Title/Summary/Keyword: Protein stability

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High-level Expression and Characterization of the Human Interleukin-10 in the Milk of Transgenic Mice

  • Zneng, Z. Y.;B. H. Sohn;K. B. Oh;W. J. Shin;Y. M. Han;Lee, K. K.
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.46-46
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    • 2003
  • Interleukin-10 (IL-10) is a homodimeric protein with a wide spectrum of anti-inflammatory and immune activities. It inhibits cytokine production and expression of immune surface molecules in various cell types. The transgenic mice carrying the human IL-10 gene in conjunction with the bovine $\beta$-casein promoter produced the human IL-10 in milk during lactation. Transgenic mice were generated using a standard method as described previously. To screen transgenic mice, PCR was carried out using chromosomal DNA extracted from tail or toe tissues with a primer set. In this study, stability of germ line transmission and expression of IL-10 gene integrated into host chromosome were monitored up to generation F15 of a transgenic line. When female mouse of generation F9 was crossbred with normal male, generation F9 to F15 mice showed similar transmission rates (66.0$\pm$20.13%, 61.5$\pm$16.66%, 41.1$\pm$8.40%, 40.7$\pm$20.34%, 61.3$\pm$10.75%, 49.2$\pm$18.82%, and 43.8$\pm$25.91%, respectively), implying that the IL-10 gene can be transmitted stably up to long term generation in the transgenic mice. For ELISA analysis, IL-10 expression levels were determined with an hIL-10 ELISA and a mIL-10 ELISA kit in accordance with the supplier's protocol. Expression levels of human IL-10 from milk of generation F9 to F13 mice were 3.6$\pm$1.20 mg/ml, 4.2$\pm$0.93 mg/ml, 5.7$\pm$1.46 mg/ml, 6.3$\pm$3.46 mg/ml, and 6.8$\pm$4.52 mg/ml, respectively. These expression levels are higher than in generation F1 (1.6 mg/ml) mice. We concluded that transgenic mice faithfully passed the transgene on their progeny and successively secreted target proteins into their milk through several generations, although there was a little fluctuation in the transmission frequency and expression level between the generations.

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Fermentation Characteristics of Large-scale Coenzyme Q10 Expressing Rhodobacter spharoides in Rumen Simulated Continuous Culture (RSCC) System (Coenzyme Q10 다량 발현 미생물을 이용한 Rumen Simulation Continuous Culture (RSCC) System 반추위 내 미생물 발효 특성에 대한 연구)

  • Bae, G.S.;Yeo, J.M.;Chang, M.B.;Kim, J.N.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.19 no.1
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    • pp.139-151
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    • 2017
  • This study was conducted to confirm the rumen fermentation characteristics of large-scale CoenzymeQ10(CoQ10) producing bacteria R. spharoides in rumen. We conducted in vitro continuous culture test to investigate the characteristics of rumen fermentation with 5% R. spharoides as a direct fed microorganism. A rumen microbial fermentation characteristic has stability at after 12 days for 15 day of experimental period. pH value, NH3-N, microbial protein synthesis, ADF digestibility and NDF digestibility were not shown significantly differences between control and treatment. However, UDP was significantly higher in treatment than control (p<0.05). CoQ10 concentration was 336.0mg/l with 5% R. spharoides. On the other hands, CoQ10 was not detected without R. spharoides. Our study was shown that R. spharoides can produce CoQ10 in rumen environment without harmful effects on rumen fermentation parameter. CoQ10 in rumen may transfer into cow milk through cow metabolism. This strategy might be helpful for producing functional dairy cow milk.

Quality Properties and Storage Characteristics of Hamburger Patty Added with Purple Kohlrabi (Brassica oleracea var. gongylodes) (자색 콜라비를 첨가한 햄버거 패티의 품질 및 저장 특성)

  • Cha, Seon-Suk;Lee, Jae-Joon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.12
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    • pp.1994-2003
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    • 2013
  • This study is performed to evaluate the effects of purple Kohlrabi addition on the quality properties and stability characteristics of hamburger patties during storage. The patties were prepared by adding 0% (N), 3.3% (K1), 6.6% (K2), and 10% (K3) of chopped purple Kohlrabi. Each patty was tested in triplicate and assigned to one of the four storage periods: 0, 5, 10, or 15 days. Addition of Kohlrabi decreased the protein and lipid contents, however, the ash and moisture contents were significantly increased. The total amino acid contents of N, K1, K2, and K3 were $15.34{\pm}1.02$, $14.57{\pm}1.28$, $15.10{\pm}1.17$, and $16.70{\pm}1.23$ mg/100 g, respectively. Palmitic acid was the most abundant among the saturated fatty acids, while oleic acid was the most abundant unsaturated fatty acids among the four groups. The water holding capacity value and cooking loss were not significantly different among the patties. In the textural characteristics, the addition of Kohlrabi increased the cohesiveness and chewiness values, but did not affect the hardness and springiness values of the patties. In the sensory evaluation, an addition of 10% Kohlrabi had the best score in color, flavor, and total acceptability. The pH of the patties decreased longer period storage; however, the total microbial counts, thiobarbituric acid (TBA) value and volatile basic nitrogen (VBN) content increased during storage. The TBA value and VBN content of the patties containing 10% Kohlrabi were lower than those of the N. Kohlrabi addition decreased the b (yellowness) and a (redness) values, and did not affect the L (lightness) value. Thus, this result suggests that adding Kohlrabi of 10% can be applied to patties for its functionality.

Extraction Conditions and Quality Stability of Carotenoprotein from Krill Processing Waste by Proteolytic Enzymes (크릴 가공폐기물을 이용한 Carotenoprotein의 추출조건 및 품질안정성에 관한 연구)

  • Kim Se-Kwon;KiM Yong-Tae;KWAK Dong-Chae;CHO Duck-Jae;LEE Eung-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.23 no.1
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    • pp.40-50
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    • 1990
  • The purpose of this paper is to develop a colorant from krill, Euphausia superba, process wastes for use in food products. Carotenoproteins were extracted from preboiled krill processing offal(PKPO) and raw frozen krill processing offal(RKPO) with the aid of proteolytic enzymes. The long-term stability of the astaxanthin associated with the carotenoprotein by the addition of pretense inhibitor and antioxidant to the product were also investigated. Total astaxanthin contents of PKPO and RKPO were $35.1mg\%,\;22.1mg\%$ and those in carotenoproteins were $98.6mg\%,\;61.9mg\%$, respectively. The chitin contents of PKPO and RKPO were $6.9\%,\;4.5\%$, however, those of carotenoproteins were not determined. When $0.5\%$ trypsin was added to the extraction medium containing 0.5M $Na_3EDTA$ at $4^{\circ}C,\;74\%$ of astaxanthin and $83\%$ of the protein of PKPO were recovered as carotenoprotein in 24hrs. The amino acid profile in carotenoprotein was mainly composed of glutamic acid, methionine, aspartic acid and isoleurine. Their contents amounted to about 40% of the total amino acids, followed by alanine, phenylalanine, Iysine, leucine, threonine and tyrosine in that order, with a small amount of cysteine and tryptophan. The levels of essential amino acids were high as much as $38.3\%\~43.6\%$ of the total amino acids. The maximum observance of the carotenoid fraction from krill processing offal and from carotenoprotein was 469nm in petroleum ether. The separated components of carotenoprotein by TLC had Rfs $0.20\~0.23\;0.56\~0.60$ and $0.88\~0.91$. The carotenoids were comprised of astaxanthin, astaxanthin monoester and asthaxanthin diester in $25\~30\%\;,35\~40\%$and $40\~45\%$, respectively. The loss of carotenoids in the carotenoprotein can be prevented by the addition of pro-tease inhibitor(trasylol) and antioxidant(BHT) below $4^{\circ}C$.

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Production of Amylase by a Thermophi1ic Fungus, Mucor Sp. (고온성(高溫性) 사상균(絲狀菌) Mucor Sp.에 의(依)한 Amylase의 생산(生産))

  • Lee, Sang Ho;Park, Yoon Joong
    • Korean Journal of Agricultural Science
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    • v.15 no.2
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    • pp.153-163
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    • 1988
  • This experiment was carried out to obtain the thermophilic fungus producing amylase and to investigate properties of the amylase. The selected strain, L-11 was obtained from soil in the vicinity of a hot spring and identified as Mocor sp.. And then the conditions for enzyme production in wheat bran cultures and properties of the crude enzyme were investigated. Furthermore, the enzyme was purified and the characteristics of purified enzyme were studied. The results obtained were as follows: 1. On the wheat bran medium added 80-100% water, amylase was effectively produced by the selected strain, L-11 for 48 hrs incubation at $50^{\circ}C$. 2. When the crude enzyme solution of the strain L-11 was passed through DEAE-Sephadex A-50 column chromatography, two peaks having amylase activity were obtained, and one peak was that of the main enzyme (enzyme of B peak). 3. The purified enzyme (enzyme of B peak) was recognized as single protein band on polyacrylamide disc gel electrophoresis. 4. In the hydrolysis reaction of soluble starch by the enzyme of main amylase, oligosaccharides produced at early stage were maltose and maltotriose mainly and procedure of the reaction maltose amount of maltose and glucose was increased. 5. The strain L-11 was recognized as a special strain producing ${\alpha}-amylase$ mainly and scarcely glucoamylase. 6. The optimum pH, optimum temperature, pH stability, and temperature stability of ${\alpha}-amylase$ were pH 4.0, $60-65^{\circ}C$, pH 4.0-9.0, and below$70^{\circ}C$.

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Effects of Different Cultivars and Milling Degrees on Quality Characteristics of Barley Makgeolli (보리의 품종 및 도정률이 막걸리의 품질 특성에 미치는 영향)

  • Park, Hye-Young;Choi, Induck;Oh, Sea Kwan;Woo, Koan Sik;Yoon, Soon Duck;Kim, Hyun-Joo;Sim, Eun-Yeong;Jeong, Seok Tae
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.12
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    • pp.1839-1846
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    • 2015
  • The purpose of this study was to evaluate the effects of cultivar and milling degree of barley on quality characteristics of Makgeolli as well as compare differences between barley Makgeolli and rice Makgeolli. Saessal-bori groups (Ss-4, Ss-12, and Ss-18) showed dry lees contents of 92.3 g, 69.4 g, and 63.8 g, respectively, whereas Huinchalssal-bori groups (Hcs-6, Hcs-14, and Hcs-20) showed contents of 62.3 g, 42.2 g, and 32.2 g. There were significant differences in quality characteristics between milling degrees and cultivars (P<0.05). The moisture, ash, crude protein, and crude fat contents of raw materials decreased with elevated milling degree. Especially, ash content of raw materials had a direct effect on Makgeolli. The pH, total acidity, and amino acidity milling with elevated decreased degree. There were no differences in total sugar or alcohol content in Makgeolli according to milling degree of barley, whereas there were significant differences between cultivars. Barley Makgeolli showed total sugar and alcohol contents of $10.7{\sim}11.8^{\circ}Brix$ and 14.07~15.07%, respectively, which were significantly lower than $12.0{\sim}12.2^{\circ}Brix$ and 17.27~17.77% in rice Makgeolli (P<0.05). Differences in colors of raw barley according to milling degree had effects on chromaticity of Makgeolli; as milling degree increased, L and b values increased. Lactic acid bacteria counts were 7.21, 6.99, and 6.67 log CFU/mL in Ss-4, Ss-12, and Ss-18, respectively, as well as 6.14, 5.39, and 5.65 log CFU/mL in Hcs-6, Hcs-14, and Hcs-20, which suggests significant reductions with increased milling degree (P<0.05). The same trend was observed in yeast as a key quality of Makgeolli. Suspension stability differed depending on milling degree, so it is expected that suspension stability can be improved by adjusting milling degree.

Processing of Ready-to-Cook Food Materials with Dark Fleshed Fish 1, Processing of Ready-to-Cook Sardine Meat "Surimi" (일시다획성 적색육어류를 이용한 중간식품소재 개발에 관한 연구 1. 정어리 연육의 가공)

  • LEE Byeong-Ho;LEE Kang-Ho;YOU Byeong-Jin;SUH Jae-Soo;JEONG In-Hak;JUNG Woo-Jin;KANG Jeong-Oak
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.18 no.5
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    • pp.401-408
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    • 1985
  • In order to develop new types of product which can offer a sanitary and preservative duality, and convenience to consumers in marketing and cooking particularly in urban area, two processing methods of ready-to-cook food materials with dark fleshed fishes like sardine and mackerel were investigated. A method applied, in this work, is processing of ready-to-cook sardine meat "surimi" in which sardine meat is treated with alkaline solution to stabilize myofibrillar proteins, washed thoroughly with water to remove soluble components, and added with a proper amount of polyphosphate and sorbitol to enforce the functional property of meat such as water holding capasity, elasticity, and gel strength. The textural properties of fish meat paste made from the "surimi" meat were greatly dependent upon the stability of myofibrillar proteins and the elimination of water soluble components. The salt soluble proteins of sardine meat were so unstable in post-mortem stage that the gel forming ability was lost within 3 days at $5^{\circ}C$ storage and 2 to 3 weeks even at $-20^{\circ}C$ although the freshness was well kept for a week at $5^{\circ}C$ and several months of storage at $-20^{\circ}C$. A proper way of treatment to keep the proteins stable was that fish meat must be washed with $0.4\%$ sodium bicarbonate solution followed by 3 to 4 times washing with water. This resulted in removal of $80\%$ water soluble proteins and 50 to $60\%$ lipids. The addition of polyphosphate and sorbitol affected the stability of proteins during the storage of "surimi" meat. When phosphate and sorbitol were added in the ratio of $0.3\%:\;0.3\%,\;0.6\%:\;3\%,\;0.6\%:\;6\%,\;0:\,0.3\%\;and\;0.3\%:\;0$, the gel forming ability terminated in 35 days, 21 days, 14 days, 14 days, and 14 days of storage at $-30^{\circ}C$, respectively, while that of the control was 7 days. And it was also noteworthy that at least 8.0 mg/g of salt soluble protein nitrogen content was required for gel formation.

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Destabilization of TNF-α mRNA by Rapamycin

  • Park, Jong-Woo;Jeon, Ye-Ji;Lee, Jae-Cheol;Ahn, So-Ra;Ha, Shin-Won;Bang, So-Young;Park, Eun-Kyung;Yi, Sang-Ah;Lee, Min-Gyu;Han, Jeung-Whan
    • Biomolecules & Therapeutics
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    • v.20 no.1
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    • pp.43-49
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    • 2012
  • Stimulation of mast cells through the high affinity IgE receptor (Fc${\varepsilon}$RI) induces degranulation, lipid mediator release, and cytokine secretion leading to allergic reactions. Although various signaling pathways have been characterized to be involved in the Fc${\varepsilon}$RI-mediated responses, little is known about the precious mechanism for the expression of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) in mast cells. Here, we report that rapamycin, a specific inhibitor of mammalian target of rapamycin (mTOR), reduces the expression of TNF-${\alpha}$ in rat basophilic leukemia (RBL-2H3) cells. IgE or specific antigen stimulation of RBL-2H3 cells increases the expression of TNF-${\alpha}$ and activates various signaling molecules including S6K1, Akt and p38 MAPK. Rapamycin specifically inhibits antigeninduced TNF-${\alpha}$ mRNA level, while other kinase inhibitors have no effect on TNF-${\alpha}$ mRNA level. These data indicate that mTOR signaling pathway is the main regulation mechanism for antigen-induced TNF-${\alpha}$ expression. TNF-${\alpha}$ mRNA stability analysis using reporter construct containing TNF-${\alpha}$ adenylate/uridylate-rich elements (AREs) shows that rapamycin destabilizes TNF-${\alpha}$ mRNA via regulating the AU-rich element of TNF-${\alpha}$ mRNA. The antigen-induced activation of S6K1 is inhibited by specific kinase inhibitors including mTOR, PI3K, PKC and $Ca^{2+}$chelator inhibitor, while TNF-${\alpha}$ mRNA level is reduced only by rapamycin treatment. These data suggest that the effects of rapamycin on the expression of TNF-${\alpha}$ mRNA are not mediated by S6K1 but regulated by mTOR. Taken together, our results reveal that mTOR signaling pathway is a novel regulation mechanism for antigen-induced TNF-${\alpha}$ expression in RBL-2H3 cells.

Development and Evaluation of the Supplementary Foods for Korean Infants and Children (지역 식품을 이용한 이유 보충식의 개발과 이의 영양학적 검토 및 저장성에 관한 연구)

  • Ahn, Kyung-Mi;Moon, Soo-Jae;Choi, Hong-Sik;Kwak, Tong-Kyung
    • Journal of Nutrition and Health
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    • v.18 no.4
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    • pp.259-271
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    • 1985
  • The purpose of this study was to develop supplementary foods for infants and young children in order to improve their nutritional status. Three formulas composed of rice, soybeans, fish, dry skim milk and sesame in varying proportions were studied. The three formulas, $RS_{1}S_{2}$, $RFS_{1}S_{2}$, and $RMS_{1}S_{2}$, were consisted of Rice(R), Soybean$(S_{1})$, Sesame$(S_{2})$ (60 : 35 : 5) , Rice, Fish(F), Soybean, Sesame (60 : 10 : 25 : 5) , and Rice, Dry Skin Milk (M), Soybean, Sesame (60 : 10 : 25 : 5), respectively. A proximate analysis and amino acid determination were made on the developed formulas. In the animal assay, growth rate, PER and FER were evaluated and biochemical analyses were also carried out. A storage test and the cost evaluation were also conducted. The summarized results are as follows : 1) The proximate composition of the three formulas were 7.3-7.4% of moisture, 15.9-21.5% of crude protein, 7.8-9.6% of crude fat and 2.5-2.8% ash. 2) The result of amino acid analysis showed that the 1st limiting amino acids of $RS_{1}S_{2}$ and $RFS_{1}S_{2}$ were lysine (amino acid score, 76.6) and threonine (amino acid score, 93.3), and that of $RMS_{1}S_{2}$ and the commercially prepared formula were sulfur containing amino acids (amino acid score, 82.0 and 54.4). When the contents of the amino acids of the three formulas were compared with mother's milk and cow's milk, the balance of the amino acid of each formula was superior to mother's milk but inferior to cow's milk. 3) In the animal assay, the growth rate of all groups increased gradually during the experimental period. 4) The C- PER, which was corrected on the basis of the casein PER of 2.5 was 2.99, 3.38 and 3.10 in the $RS_{1}S_{2}$, $RFS_{1}S_{2}$ and $RMS_{1}S_{2}$ respectively. The C- PER of $RFS_{1}S_{2}$ and $RMS_{1}S_{2}$ were Significantly (P<.05) higher than that of the casein. 5) The FER of the casein, $RS_{1}S_{2}$, $RFS_{1}S_{2}$, and $RMS_{1}S_{2}$ were 0.37, 0.39, 0.43 and 0.39, respectively. The FER of $RFS_{1}S_{2}$ and $RMS_{1}S_{2}$ were also significantly (P<.05) higher than that of the casein. 6) The concentrations of hematocrit, hemoglobin, total protein and albumin in the serum of the rats of all groups were not significantly different among groups. 7) The storage stability test showed that the total plate count (TPC), the coliforms count and the bacterial spore count in the ingredients were quiet low. However, after 30 and 60 days storage, the count in $RFS_{1}S_{2}$ increased and were higher at room temperature than refrigerated temperature. 8) In the cost evaluation, the cost of the developed formulas was \1,826-2,626 / kg. This was less than that of the commercially prepared formula (\3,300-4,073 / kg) and that of the imported formula (\4,250-8,720 / kg).

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Physicochemical Properties of Commercial Sweet Potato Starches (시판 고구마전분의 이화학적 특성)

  • Baek, Man-Hee;Cha, Dong-Su;Park, Hyun-Jin;Lim, Seung-Taik
    • Korean Journal of Food Science and Technology
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    • v.32 no.4
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    • pp.755-762
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    • 2000
  • Physicochemical properties of commercial sweet potato starches manufactured by 7 different companies were investigated in comparison with corn and potato starches. Crude ash and protein content varied from 0.36 to 1.02%, and from 0.04 to 0.14% based on dry weight, respectively. The protein contents were relatively smaller than that of corn or potato starch. But whiteness of the sweet potato starches was less than that of corn or potato starch. Mean diameter of the sweet potato starch granules varied from 14.23 to $21.08\;{\mu}m$ depending on the company and all sweet potato starches showed bimodal size distributions. Pasting viscosity measured by Rapid Viscoanalyzer(RVA) also showed variations among the starches of different companies. The starch from D company in Korea had the lowest pasting temperature$(74.00^{\circ}C)$ whereas the starch from a phillippine company(P) did the highest one$(80.35^{\circ}C)$. The peak viscosity of sweet potato starches was higher than that of corn starch but lower than that of potato starch. The D company starch also showed the highest peak viscosity(2283 cp) among the starches tested. Paste breakdown by hot shearing ranged from 524 cp (S company) to 1279 cp (HL company). Textural properties of the starch gels appeared significantly different among the starches of different manufacturers. The greatest hardness of the gel was $137.90\;g_{f}$ at 1 day storage whereas the lowest value was $31.53\;g_{f}$. Except the starches from 2 companies (P and S), the sweet potato starches formed very soft and weak gels. P or S company starches formed the gels similar to potato starch. Syneresis by freeze-thawing treatments appeared less for sweet potato starch gels than that for corn starch gels, but greater than that for potato starch gel. The overall properties of the sweet potato starches varied by the manufacturing companies, and ranged between those of corn and potato starches.

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