• 제목/요약/키워드: In Vitro Protein Solubility

검색결과 33건 처리시간 0.028초

Relationship between Molecular Structure Characteristics of Feed Proteins and Protein In vitro Digestibility and Solubility

  • Bai, Mingmei;Qin, Guixin;Sun, Zewei;Long, Guohui
    • Asian-Australasian Journal of Animal Sciences
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    • 제29권8호
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    • pp.1159-1165
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    • 2016
  • The nutritional value of feed proteins and their utilization by livestock are related not only to the chemical composition but also to the structure of feed proteins, but few studies thus far have investigated the relationship between the structure of feed proteins and their solubility as well as digestibility in monogastric animals. To address this question we analyzed soybean meal, fish meal, corn distiller's dried grains with solubles, corn gluten meal, and feather meal by Fourier transform infrared (FTIR) spectroscopy to determine the protein molecular spectral band characteristics for amides I and II as well as ${\alpha}$-helices and ${\beta}$-sheets and their ratios. Protein solubility and in vitro digestibility were measured with the Kjeldahl method using 0.2% KOH solution and the pepsin-pancreatin two-step enzymatic method, respectively. We found that all measured spectral band intensities (height and area) of feed proteins were correlated with their the in vitro digestibility and solubility ($p{\leq}0.003$); moreover, the relatively quantitative amounts of ${\alpha}$-helices, random coils, and ${\alpha}$-helix to ${\beta}$-sheet ratio in protein secondary structures were positively correlated with protein in vitro digestibility and solubility ($p{\leq}0.004$). On the other hand, the percentage of ${\beta}$-sheet structures was negatively correlated with protein in vitro digestibility (p<0.001) and solubility (p = 0.002). These results demonstrate that the molecular structure characteristics of feed proteins are closely related to their in vitro digestibility at 28 h and solubility. Furthermore, the ${\alpha}$-helix-to-${\beta}$-sheet ratio can be used to predict the nutritional value of feed proteins.

단백질 함량 및 철분 농도의 변화에 따라 홍차 추출물이 철분의 용해도에 미치는 영향 (Effects of Protein and Iron Concentrations on Iron Solubility in Black Tea Infusion)

  • 김희선
    • Journal of Nutrition and Health
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    • 제29권8호
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    • pp.861-866
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    • 1996
  • Tannins in plant foods and beverages may produce antinutritional or toxic effects although some proteins with high affinity for tannins seem to function as defense mechanism to tannin toxicity. Our objectives were to investigate of tea tannins, iron and protein and to evaluate the role of proteins in tannin effects on iron solubility. Iron solubility in vitro was measured using tea with and without proteins. Mixtures of tea, protein in varying concentrations(either gelatin or bovine serum albumin), and iron(eithe 10 or 50ug/mL) were prepared. Controls contained water in place of tea. Iron bioavailability was assessed by measuring iron solubility in the simulated gastric condition with pepsin digestion. Bound iron was removed by centrifugation and soluble in tea alone. When iron concentratin was 10ug/mL, addition of small amounts of protein to tea dramatically reduced iron solubility, but solubility of iron increased in the tea mixturea as the concentration of protein was increased. The percnetage of iron that precipitated was much greater at 10ug Fe/mL than the values at 50ug Fe/mL suggesting that the iron binding sites on the tea-protein complex was saturated. These results suggest that interactions of iron with tea tannins are influenced by the concentratins of protein and iron.

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Predicting In Sacco Rumen Degradation Kinetics of Raw and Dry Roasted Faba Beans (Vicia faba) and Lupin Seeds (Lupinus albus) by Laboratory Techniques

  • Yu, P.;Egan, A.R.;Leury, B.J.
    • Asian-Australasian Journal of Animal Sciences
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    • 제13권10호
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    • pp.1377-1387
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    • 2000
  • Two laboratory techniques: (1) an in vitro method with two procedures for measuring protein degradabilities and (2) an in vitro method with three procedures for measuring protein solubility, were investigated to determine which laboratory techniques could most accurately predict the quantity of rumen protein degradation kinetics of legume seeds after dry roasting under various conditions, in terms of (1) rumen protein disappearance ($D_j$, where j=0, 2, 4, 8, 12, 24 and 48 h incubation), (2) rumen protein effective degradability (EDCP), (3) the parameters describing rumen degradation characteristics (the soluble fraction: S, the potentially degradable fraction: D, undegradable fraction: U, lag time: T0 and the degradation rate: Kd) and (4) rumen bypass protein (BCP), which were determined by the method accepted internationally at present, in sacco nylon bag technique using the standardized Dutch method. Feeds evaluated were the raw and dry roasted whole faba (Vicia faba) beans (WFB) and whole lupin (Lupinus albus) seeds (WLS), each was dry roasted under various conditions (at 110, 130 or $150^{\circ}C$ for 15, 30 or 45 min). In vitro protein degradability ($D_1$_Auf and $D_{24}$_Auf) were determined using the modified Aufr re method by enzymatic hydrolysis for 1 h and 24 h using a protease extracted from Streptomyces griseus in a borate-phosphate buffer. In vitro protein solubility ($bf_1$_S, $bf_2$_S, $bf_3$_S) was measured in a borate-phosphate buffer with three different procedures. Results from laboratory techniques (in vitro) were correlated and linearly regressed with in sacco results. Of the three procedures of in vitro protein solubility evaluated, none of them could predict in sacco results with good precision. The highest Pearson correlation coefficient ($R^2$) was less than 0.50. Of two procedures of in vitro protein degradability studied, the $D_1$_Auf values were closely correlated with in sacco parameters: Kd, EDCP and %BCP with high R' values: 0.82, 0.85 and 0.85, respectively, and closely correlated with in sacco $D_j$ at 2, 4, 8 and 12 h rumen incubation with high $R^2$ values: 0.83, 0.91, 0.93 and 0.83, respectively. The $D_{24}$_Auf values could not predict in sacco results. The highest $R^2$ value was less then 0.40. These results indicated that in vitro protein solubility measured in borate-phosphate failed to identify differences in the rate and extent of protein degradation of legume seeds after dry roasting under various conditions and thus should not be used to predict rumen degradation, particularly for heat processed feedstuffs. But in vitro protein degradability using the modified Aufr re method by enzymatic hydrolysis for 1 h or possibly an intermediate time (>1 h and <24 h) is a promising laboratory procedure to detect effectiveness of dry roasting legume seeds on rumen protein degradation characteristics and could be used as a simple laboratory method to predict the rate and extent of protein degradation in the rumen in sacco with high accuracy. The equations to predict EDCP, Kd and BCP of dry roasted legume seeds (WLS and WFB) under various conditions are as follow: For both: EDCP (%)=-1.37+1.06*$D_1$_Auf ($R^2=0.85$, p<0.01). For both: Kd (%/h)=-21.81+0.49*$D_1$_Auf ($R^2=0.82$, p<0.01). For both: %BCP=103.37-1.07*$D_1$_Auf ($R^2=0.85$, p<0.01).

분리 유채단백의 용해도와 소화율에 미치는 Phytate의 영향 (Effect of Phytate on the Solubility and Digestibility of Rapeseed Protein Isolate)

  • 조희경;윤재영;이서래
    • 한국식품과학회지
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    • 제24권3호
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    • pp.279-283
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    • 1992
  • 식물성 식품중의 영양 저해 인자로 알려진 phytate를 제거한 분리 유채단백(low-phytate rapeseed protein isolate)을 제조하여 pH 및 phytate 첨가량이 이들 분리 유채단백의 용해성과 소화율에 미치는 영향을 살펴보았다. 분리 유채단백의 phytate 함량은 1.5%로서 탈지 유채분말보다 66%가 감소되었으며 protein : phytate ratio는 58 : 1이었다. 분리 유채단백의 용해도는 pH 2.0과 pH 11.5에서는 매우 높았으나 pH 5.0에서는 매우 낮았다. 그 용해도에 미치는 phytate 첨가량의 영향을 보면 pH 5.0과 pH 11.5에서는 별 차이가 없었으나 pH 2.0에서는 phytate 첨가량이 많을수록 그 용해도가 감소하였다. 분리 유채단백의 pepsin 소화율에 미치는 phytate의 저해적 영향은 phytate 함량이 증가할수록 더 커졌다. 소화기간 초기에는 그 영향이 매우 컸으나 소화시간이 길어짐에 따라 그 영향은 점점 감소되었으며 평균소화율은 24%가 감소하였다. 따라서 유채단백을 식품소재로 이용하기 위해서는 phytate 함량이 낮은 제품을 제조함으로써 그의 기능성과 영양가를 향상시킬 수 있을 것이다.

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동결건조 어육의 단백질 Cross-linking에 관한 연구 (Protein Cross-linking in Freeze Dried Fish Meat)

  • 이강호;정우진;서재수;정인학;김충곤
    • 한국수산과학회지
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    • 제19권2호
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    • pp.100-108
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    • 1986
  • This study was carried out in order to investigate protein cross-linking in freeze-dried meat of flounder (Limanda herzensteini). Changes in solubility or extractability of proteins and electrophoretic patterns of the extracted proteins were determined to monitor the cross-linking during the storage of freeze-dried meat. Development of nonenzymatic browning and the loss of in vitro protein digestibilily were also measured to assess their influences on the changes of functional and nutritional properties of proteins. In addition, the effects of lysine added, and removal of fat and water extractives were also mentioned. The extractability of protein decreased upon storage time and temperature, and the loss of solubility of myosin was evident. In case of the samples stored at $5^{\circ}C$ for 150 days, the extractability of protein decreased $26.4\%$, while that of the samples stored at $20^{\circ}C$ for 60 days decreased about $39.7\%$. And it was noted that the loss of solubility of myosin was $68.3\%$ and $98.1%$ for the same storage conditions, respectively. It was noteworthy that the samples treated with $L-lysine{\cdot}HCl$ seemed to prevent more or less the loss of protein solubility, in that, even stored at $20^{\circ}C$ for 120 days, revealed only $57.03\%$ decrease. The nonenzymatic browning was proceeded with the increase of storage temperature, especially, in the samples treated with glucose. This suggests that the decrease in extractibility of myosin was accompanied by the extent of browning. But the browning was retarded in defatted samples. The in vitro apparent protein digestibility was also higher in the samples defatted or water extracted. It was suggested from these results that changes in properties of proteins in freeze dried fish meat were led by the protein cross-linking which was attributed to Maillard type of reactions and protein-lipid interactions.

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Effects of Extrusion Variable on Functional and Nutritional Properties of Extruded Oat Products

  • Gutkoski, Luiz Carlos;El-Dash, Ahmed Atia
    • Preventive Nutrition and Food Science
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    • 제4권3호
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    • pp.159-162
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    • 1999
  • The purpose of this research was to study the effects of initial moisture levels and extrusion temperatures on dietary fiber, nitrogen solubility index, available lysine, and the in vitro protein digestibility of extruded oat productes. The dehulled grains were ground in a Brabender quadrumat Senior mill and the coarse fraction, with higher crude protein, lipids and dietary fiber were conditioned on various mositre levels (15.5~25.5%) and extruded in a Brabender single-screw laboratory extruder. The extrudates showed a higher amount of soluble dietary fiber (8.14%) than in the raw material . However, the extrusion process affected the nutritional value of the protein due to a decrease in available lysine with increased temperature . The in vitro protein digestibility was unaffected by initial moisture levels and the extrusion temperatures examined.

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EFFECT OF HEAT TREATMENT ON NUTRITIONAL VALUE OF WINGED BEAN (Psophocarpus tetragonolobus) AS COMPARED TO SOYBEAN I. CHEMICAL CHARACTERISTICS OF TREATED WINGED BEAN

  • Mutia, R.;Uchida, S.
    • Asian-Australasian Journal of Animal Sciences
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    • 제6권1호
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    • pp.19-26
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    • 1993
  • The effect heat treatment (autoclave) on nutritional value of winged bean as compared to soybean has been investigated. The winged bean and soybean were obtained from local cultivar grown in Indonesia. The beans were autoclaved at $120^{\circ}C$ for 15, 30, 45, 60 or 90 minutes, respectively before being ground for chemical analysis. Trypsin inhibitors of winged bean and soybean decreased (p < 0.05) along with decreasing of urease activity as heating time increased from 0 to 90 minutes. Heat treatment significantly (p < 0.05) reduced protein solubility in 0.2% potassium hydroxide of winged bean as well as soybean. In vitro protein digestibility was significantly (p < 0.05) improved by heating treatment (15 to 60 min of autoclaving), however, excessive heating (90 min of autoclaving) decreased the digestibility of winged beans. Excessive heating had adverse effect on lysine, cystine and methionine contents of winged beans. The results of this study suggested that autoclaving at $120^{\circ}C$ within 45 minutes should be adequate to remove protease inhibitors and could improve protein digestibility of winged beans.

천연 밀단백질/계면활성제 복합체의 세정에 있어 피부보호 (The Cutaneous Protection for Detergent Formulation of Nature Wheat Protein Surfactant Complexes)

  • 정환경;박흥조;김명수;남기대
    • 한국응용과학기술학회지
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    • 제19권2호
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    • pp.79-85
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    • 2002
  • The cutaneous tolerability of detergent formulations can be improved by means of suitable additives. They complex the surfactant molecules lowering the concentration of their free monomeric species. Proteins derivatives used as additives for detergency are usually prepared by partial hydrolysis of plant reserve proteins. The main purpose of the hydrolytic cleavage is to make them water soluble and suitable for liquid products. Water solubility and stability are obtained by means of complexation with surfactants which also increase their actual hydrophobicity, an important parameter affecting cosmetic properties of proteins. Transepidermal water loss (TEWL) and electric capacitance (EC) have been adopted as investigation technigues to evaluate the skin integrity/damage in vitro tests, The performance of native wheat protein / surfactant complexes has been compared with traditional protein hydrolysates as detergent additives. The results show a noticeable reduction of skin irritation in surfactant formulations with addition of native wheat protein.

Construction, Investigation and Application of TEV Protease Variants with Improved Oxidative Stability

  • Bayar, Enkhtuya;Ren, Yuanyuan;Chen, Yinghua;Hu, Yafang;Zhang, Shuncheng;Yu, Xuelian;Fan, Jun
    • Journal of Microbiology and Biotechnology
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    • 제31권12호
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    • pp.1732-1740
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    • 2021
  • Tobacco etch virus protease (TEVp) is a useful tool for removing fusion tags, but wild-type TEVp is less stable under oxidized redox state. In this work, we introduced and combined C19S, C110S and C130S into TEVp variants containing T17S, L56V, N68D, I77V and S135G to improve protein solubility, and S219V to inhibit self-proteolysis. The solubility and cleavage activity of the constructed variants in Escherichia coli strains including BL21(DE3), BL21(DE3)pLys, Rossetta(DE3) and Origami(DE3) under the same induction conditions were analyzed and compared. The desirable soluble amounts, activity, and oxidative stability were identified to be reluctantly favored in the TEVp. Unlike C19S, C110S and C130S hardly impacted on decreasing protein solubility in the BL21(DE3), but they contributed to improved tolerance to the oxidative redox state in vivo and in vitro. After two fusion proteins were cleaved by purified TEVp protein containing double mutations under the oxidized redox state, the refolded disulfide-rich bovine enterokinase catalytic domain or maize peroxidase with enhanced yields were released from the regenerated amorphous cellulose via affinity absorption of the cellulose-binding module as the affinity tag.

Protection of Mice Against Pandemic H1N1 Influenza Virus Challenge After Immunization with Baculovirus-Expressed Stabilizing Peptide Fusion Hemagglutinin Protein

  • Yang, Eunji;Cho, Yonggeun;Choi, Jung-ah;Choi, YoungJoo;Park, Pil-Gu;Park, Eunsun;Lee, Choong Hwan;Lee, Hyeja;Kim, Jongsun;Lee, Jae Myun;Song, Manki
    • Journal of Microbiology and Biotechnology
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    • 제25권2호
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    • pp.280-287
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    • 2015
  • Current influenza vaccines are produced in embryonated chicken eggs. However, egg-based vaccines have various problems. To address these problems, recombinant protein vaccines have been developed as new vaccine candidates. Unfortunately, recombinant proteins frequently encounter aggregation and low stability during their biogenesis. It has been previously demonstrated that recombinantly expressed proteins can be greatly stabilized with high solubility by fusing stabilizing peptide (SP) derived from the C-terminal acidic tail of human synuclein (ATS). To investigate whether SP fusion proteins can induce protective immunity in mice, we produced influenza HA and SP fusion protein using a baculovirus expression system. In in vitro tests, SP-fused recombinant HA1 (SP-rHA1) was shown to be more stable than recombinant HA1 (rHA1). Mice were immunized intramuscularly with baculovirus-expressed rHA1 protein or SP-rHA1 protein ($2{\mu}g/mouse$) formulated with aluminum hydroxide. Antibody responses were determined by ELISA and hemagglutination inhibition assay. We observed that SP-rHA1 immunization elicited HA-specific antibody responses that were comparable to rHA1 immunization. These results indicate that fusion of SP to rHA1 does not negatively affect the immunogenicity of the vaccine candidate. Therefore, it is possible to apply SP fusion technology to develop stable recombinant protein vaccines with high solubility.