• 한국축산식품학회지
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• 제37권6호
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• pp.940-947
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• 2017

Goat milk has a protein composition similar to that of breast milk and contains abundant nutrients, but its use in functional foods is rather limited in comparison to milk from other sources. The aim of this study was to prepare a goat A2 ${\beta}$-casein fraction with improved digestibility and hypoallergenic properties. We investigated the optimal conditions for the separation of A2 ${\beta}$-casein fraction from goat milk by pH adjustment to pH 4.4 and treating the casein suspension with calcium chloride (0.05 M for 1 h at $25^{\circ}C$). Selective reduction of ${\beta}$- lactoglobulin and ${\alpha}_s$-casein was confirmed using sodium dodecyl sulphate-polyacrylamide gel electrophoresis and reverse-phase high-performance liquid chromatography. The hypoallergenic property of A2 ${\beta}$-casein fraction was examined by measuring the release of histamine and tumor necrosis factor alpha from HMC-1 human mast cells exposed to different proteins, including A2 ${\beta}$-casein fraction. There was no significant difference in levels of both indicators between A2 ${\beta}$-casein treatment and the control (no protein treatment). The A2 ${\beta}$-casein fraction is abundant in essential amino acids, especially, branched-chain amino acids (leucine, valine, and isoleucine). The physicochemical properties of A2 ${\beta}$-casein fraction, including protein solubility and viscosity, are similar to those of bovine whole casein which is widely used as a protein source in various foods. Therefore, the goat A2 ${\beta}$-casein fraction may be useful as a food material with good digestibility and hypoallergenic properties for infants, the elderly, and people with metabolic disorders.

• Asian-Australasian Journal of Animal Sciences
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• 제22권4호
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• pp.534-541
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• 2009

The aim of this study was to evaluate the effects of gamma irradiation (${\gamma}$-irradiation) at doses of 15, 30 and 45 kGy on chemical composition, anti-nutritional factors, ruminal dry matter (DM) and crude protein (CP) degradibility, in vitro CP digestibility and to monitor the fate of true proteins of full-fat soybean (SB) in the rumen. Nylon bags of untreated or ${\gamma}$-irradiated SB were suspended in the rumens of three ruminally-fistulated bulls for up to 48 h and resulting data were fitted to a nonlinear degradation model to calculate degradation parameters of DM and CP. Proteins of untreated and treated SB bag residues were fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Digestibility of rumen undegraded CP was estimated using the three-step in vitro procedure. The chemical composition of raw and irradiated soybeans was similar. Results showed that phytic acid in ${\gamma}$-irradiated SB at dose of 30 kGy was eliminated completely. The trypsin inhibitor activity of 15, 30 and 45 kGy ${\gamma}$-irradiated SB was decreased (p<0.01) by 18.4, 55.5 and 63.5%, respectively. From in sacco results, ${\gamma}$-irradiation decreased (p<0.05) the washout fractions of DM and CP at doses of 30 and 45 kGy, but increased (p<0.05) the potentially degradable fractions. Gamma irradiation at doses of 15, 30 and 45 kGy decreased (p<0.05) effective degradability of CP at a rumen outflow rate of 0.05 $h^{-1}$ by 4.4, 14.4 and 26.5%, respectively. On the contrary, digestibility of ruminally undegraded CP of irradiated SB at doses of 30 and 45 kGy was improved (p<0.05) by 12 and 28%, respectively. Electrophoretic analysis of untreated soybean proteins incubated in the rumen revealed that ${\beta}$-conglycinin subunits had disappeared at 2 h of incubation time, whereas the subunits of glycinin were more resistant to degradation until 16 h of incubation. From the SDS-PAGE patterns, acidic subunits of 15, 30 and 45 kGy ${\gamma}$-irradiated SB disappeared after 8, 8 and 16 h of incubation, respectively, while the basic subunits of glycinin were not degraded completely until 24, 48 and 48 h of incubation, respectively. It was concluded that ${\gamma}$-irradiated soybean proteins at doses higher than 15 kGy could be effectively protected from ruminal degradation.

• Asian-Australasian Journal of Animal Sciences
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• 제25권11호
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• pp.1540-1545
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• 2012

Steroidogenesis requires coordination of the anabolic and catabolic pathways of lipid metabolism, but the profile of proteins associated with progesterone synthesis in cyclic and pregnant corpus luteum (CL) is not well-known in cattle. In Experiment 1, plasma progesterone level was monitored in cyclic cows (n = 5) and pregnant cows (n = 6; until d-90). A significant decline in the plasma progesterone level occurred at d-19 of cyclic cows. Progesterone level in abbatoir-derived luteal tissues was also determined at d 1 to 5, 6 to 13 and 14 to 20 of cyclic cows, and d-60 and -90 of pregnant cows (n = 5 each). Progesterone level in d-60 CL was not different from those in d 6 to 13 CL and d-90 CL, although the difference between d 6 to 13 and d-90 was significant. In Experiment 2, protein expression pattern in CL at d-90 (n = 4) was compared with that in CL of cyclic cows at d 6 to 13 (n = 5). Significant changes in the level of protein expression were detected in 32 protein spots by two-dimensional polyacrylamide gel electrophoresis (2-DE), and 23 of them were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Six proteins were found only in pregnant CL, while the other 17 proteins were found only in cyclic CL. Among the above 6 proteins, vimentin which is involved in the regulation of post-implantation development was included. Thus, the protein expression pattern in CL was disorientated from cyclic luteal phase to mid pregnancy, and alterations in specific CL protein expression may contribute to the maintenance of pregnancy in Korean native cows.

• Asian-Australasian Journal of Animal Sciences
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• 제30권5호
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• pp.736-742
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• 2017

Objective: Experiments were conducted to clone the sequence of Wild Argali short palate, lung and nasal epithelium clone 1 (SPLUNC1) cDNA, and to lay the foundation for further study the biological function of Wild Argali SPLUNC1. Methods: The complete sequence of Wild Argali SPLUNC1 cDNA was generated by rapid amplification of cDNA ends. The entire coding sequence was inserted into the pPIC9K vector and expressed in Pichia pastoris (P. pastoris) GS115. The recombinant SPLUNC1 protein was detected by Western blot and purified by $Ni^{2+}$ chelate affinity chromatography. The test of effect of the protein on Mycoplasma ovipneumoniae (MO) was performed with real-time polymerase chain reaction. Results: The Wild Argali SPLUNC1 cDNA was 1,076 bp with an open reading frame of 768 bp, which encoded a 26.49 kDa protein composed of 255 amino acids. Its amino acid sequence shared 98.4%, 96.9%, 94.5%, 90.2%, 80.8%, 78.4%, 78.3%, 72.5%, 72.3%, 68.8% identity with those of SPLUNC1 cDNA from Ovis aries (accession no. NP_001288334.1), Capra hircus (accession no. XP_005688516.1), Pantholops hodgsonii (accession no. XP_005979709.1), Bos taurus (accession no. NP_776851.1), Felis catus (accession no. XP_006929910.1), Homo sapiens (accession no. NP_001230122.1), Sus scrofa (accession no. NP_001005727.1), Chinchilla lanigera (accession no. NP_001269294.1), Mus musculus (accession no. NP_035256.2), and Rattus norvegicus (accession no. NP_742028.1), respectively. The recombinant protein corresponded to the expected molecular mass of 25.47 kDa as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and it was detected in the supernatant of P. pastoris, and it could be purified. The results from the test of inhibition effect of argali recombinant SPLUNC1 protein on MO showed that the product could inhibit MO very well (p<0.01). Conclusion: The amino acid sequence of Wild Argali SPLUNC1 was different from other organisms. The recombinant SPLUNC1 protein has good biological activity.

• Asian-Australasian Journal of Animal Sciences
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• 제19권6호
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• pp.775-778
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• 2006

Sarabi cows (n = 136) from the Sarabi Breeding Station were genotyped at bovine lymphocyte antigen (BoLA)-DRB3.2 locus by a genotyping system that used the polymerase chain reaction and restriction fragment length polymorphism. Genomic DNA was extracted from whole blood samples. A two-step polymerase chain reaction was carried out in order to amplify a 284 base-pair fragment of target gene. Nested-PCR products were digested with three restriction endonuclease enzymes RsaI, BstYI and HaeIII. Digested fragments were analyzed by polyacrylamide gel electrophoresis. Twenty-six BoLA-DRB3.2 alleles were identified with frequencies ranging from 0.4 to 15.1%. Six new allele types observed in this study have not been reported previously. Identified alleles include: BoLA-DRB3.$2^*1$, $^*2$, $^*4$, $^*6$, $^*8$, $^*12$, $^*13$, $^*14$, $^*15$, $^*16$, $^*17$, $^*23$, $^*24$, $^*25$, $^*28$, $^*32$, $^*34$, $^*35$, $^*36$, $^*37$, $^*42$, $^*46$, $^*51$, $^*kba$, $^*laa$ and $^*vaa$. Their frequencies were found to be 0.4, 0.4, 0.7, 11.4, 1.1, 1.8, 2.9, 2.2, 4.4, 9.6, 1.1, 13.6, 0.4, 0.4, 1.1, 0.7, 0.4, 6.2, 2.2, 3.7, 1.1, 7.7, 1.5, 15.1, 2.6 and 7.3% respectively. The six most frequent alleles (DRB3.2 $^*6$, $^*16$, $^*23$, $^*46$, $^*kba$ and $^*vaa$) accounted for 64.7% of the alleles in the population of this herd. Numerous studies on this locus, covering different breeds, has revealed the existence of various alleles in this locus, and new investigations have introduced novel alleles. With respect to the high number of the observed alleles in this survey and the novelty of some alleles with no previous record of reporting, it is plausible to conclude that the BoLA-DRB3.2 locus is highly polymorphic in Iranian native Sarabi cows.

• Asian-Australasian Journal of Animal Sciences
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• 제29권1호
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• pp.126-133
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• 2016

A gene from Actinomyces sp. Korean native goat (KNG) 40 that encodes an endo-${\beta}$-1,4-glucanase, EG1, was cloned and expressed in Escherichia coli (E. coli) $DH5{\alpha}$. Recombinant plasmid DNA from a positive clone with a 3.2 kb insert hydrolyzing carboxyl methyl-cellulose (CMC) was designated as pDS3. The entire nucleotide sequence was determined, and an open-reading frame (ORF) was deduced. The ORF encodes a polypeptide of 684 amino acids. The recombinant EG1 produced in E. coli $DH5{\alpha}$ harboring pDS3 was purified in one step using affinity chromatography on crystalline cellulose and characterized. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis/zymogram analysis of the purified enzyme revealed two protein bands of 57.1 and 54.1 kDa. The amino terminal sequences of these two bands matched those of the deduced ones, starting from residue 166 and 208, respectively. Putative signal sequences, a Shine.Dalgarno-type ribosomal binding site, and promoter sequences related to the consensus sequences were deduced. EG1 has a typical tripartite structure of cellulase, a catalytic domain, a serine-rich linker region, and a cellulose-binding domain. The optimal temperature for the activity of the purified enzyme was $55^{\circ}C$, but it retained over 90% of maximum activity in a broad temperature range ($40^{\circ}C$ to $60^{\circ}C$). The optimal pH for the enzyme activity was 6.0. Kinetic parameters, $K_m$ and $V_{max}$ of rEG1 were 0.39% CMC and 143 U/mg, respectively.

• Fisheries and Aquatic Sciences
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• 제13권2호
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• pp.102-111
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• 2010

This study was conducted to investigate the optimal conditions of collagen extraction from scales of yellowfin tuna (Thunnus albacares) using surface response methodology. Four independent variables of NaOH concentration and pretreatment fime in alkali pretreatment and enzyme concentration and treatment time in enzyme hydrolysis were used to predict a model equation for the collagen yield. The determinant coefficient ($R^2$) for the equation was 0.906. The values of the independent variables for the maximum yield were 0.32 N NaOH, 16.38 h alkali pretreatment time, 0.18% enzyme concentration, and 31.02 h enzyme treatment time. In the physicochemical properties of tuna scale collagen, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of tuna scale collagen showed the same migration distances as that of calf skin collagen. The amide A, I, II, and III regions of tuna scale collagen in Fourier transform infrared measurements were shown in the peaks of 3,414 $cm^{-1}$, 1,645 $cm^{-1}$, 1,553 $cm^{-1}$, and 1,247 $cm^{-1}$, respectively. The amount of imino acids in tuna scale collagen was 18.97% and the collagen denaturation temperature was $33^{\circ}C$. The collagen solubility as a function of NaCl concentration decreased to 4% NaCl (w/v) and the collagen solubility as a function of pH was high at pH 2-4 and sharply decreased from pH 4 to pH 7. Viscosity of the collagen solution decreased continuously until $30^{\circ}C$ and this decreasing rate slowed in the temperature range of $35-50^{\circ}C$.

• Journal of Oral Medicine and Pain
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• 제24권2호
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• pp.125-135
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• 1999

This study was performed to discover the underlining influences of Herpes Simplex virus (HSV) and Varicella Zoster virus (VZV), to detect the changes of whole protein and mucin level and to observe protein profiles in the saliva when recurrent aphthous ulcer (RAU) was present. Unstimulated whole saliva was collected from 23 patients who for over three years had a clinical history of RAU, in a group of 10 women and 13 men, ranging from 11 to 72 years of age, and 20 healthy subjects, in a group of 8 women and 12 men, who did not have the symptoms nor a past history of RAU. Through the means of Polymerization Chain Reaction, genomic DNA from the HSV and VZV was purified from the saliva samples for identifying precisely the two types of viruses, and the level of whole protein and sialic acids in the saliva and the ratio of sialic acid to whole protein were measured, and SDS-polyacrylamide gel electrophoresis was performed. The results obtained were as follows ; 1. 39.13% of patients showed 224 bp bands of VZV DNA, those were appeared more in patients than in control group (p<0.01), but there was no significant difference between patients and control group in HSV DNA (p>0.05). 2. The concentration of whole protein in men patients was lower than in men control group (p<0.05), but there were no significant differences between patients and control in other groups (p>0.05). 3. The concentration of sialic acids from patients was lower than control group in all groups (p<0.05). 4. The concentration of sialic acids in proportion to that of whole protein was lower in patients than in control group (p<0.05), and in the two women groups (p<0.01), but no noticeable difference was found between the two men groups (p>0.05). 5. There were no consistent differences observed in the protein profiles of patients with control group except that certain protein bands near 50 kDa was lower in patients than in control group. These results suggest that viruses such as HSV and VZV and reduction of salivary whole protein and mucin levels are related to development of RAU.

• 한국약용작물학회지
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• 제14권4호
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• pp.195-201
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• 2006

미생물 및 동물에 비해 식물에서는 혈전용해효소에 대한 연구가 매우 부족한 실정이며, 기존의 혈전용해효소가 가지는 혈전에 대한 비특이적, 부작용, 고가 등의 단점을 해결할 수 있는 새로운 혈전용해효소의 개발을 위하여 동백의 종자, 종피, 유엽 그리고 성엽으로부터 추출된 수용성 단백질의 혈전용해활성을 조사하였다. 각각의 동백부위로부터 추출된 조효소 용액은 기존 혈전 용해효소인 plasmin과 양성 대조군으로 하여 비교하여 fibrin agarose plate로 확인한 결과 fibrin clot을 효과적으로 분해하였다. 그 중 단백질 분해효소의 활성이 다른 부위보다 20-33배로 높았던 동백종자와 종피의 수용성 추출물의 혈전용해활성은 양성 대조군인 plasmin과 비교하여 1.6-2.0배의 높은 활성을 나타내었다. 전체 수용성 단백질은 30-80%황산암모늄을 이용하여 농축하였으며 혈전용해효소는 fibrin zymography를 수행하여 확인하였다. SDS-PAGE에 의하여 동백유엽의 혈전용해효소 분자량을 측정한 결과 45 kDa으로 단일 polypeptide임을 확인하였으며, 각종 pretense 저해제에 의한 영향을 조사한 결과 PMSF,그리고 TLCK에 강력하게 저해되는 것으로 보아 동백유엽의 혈전용해효소는 trypsin과 유사한 serine protease의 하나로 생각되었다. 그러나 EDTA와 DTT처리에 의해서는 효소활성의 저해가 두드러지게 나타나지 않고 오히려 증진된 양상을 확인할 수 있었다. 또한, 효소 활성에 미치는 pH 및 온도의 효과는 약간의 산성쪽에 가까운 pH 5.5와 $30^{\circ}C$에서는 최적의 활성을 나타내었으며, $45^{\circ}C$ 이상의 온도에서는 효소활성이 급격히 감소하였다. 이상의 모든 결과로 볼 때 동백유엽의 혈전용해효소는 trypsin과 유사한 serine protease에 속하는 혈전용해효소임을 확인할 수 있었다.

• 한국균학회지
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• 제22권4호
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• pp.386-393
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• 1994

Fusarium 속내 종들의 분류학적 위치를 알아보기 위하여, 각 균의 $esterase-{\alpha},\;-{\beta},\;acid\;phosphatase,\;malate\;dehydrogenase,\;peroxidase,\;polygalacturonase$를 추출하여 전기영동의 방법으로 isozyme patterns를 비교하여 보았다. Banding patterns은 polygalacturonase만이 7종 모두에서 monozyme이었고, 나머지 효소는 종마다 약간씩 다르게 나타났다. Isozyme banding patterns을 바탕으로 genetic similarity를 산출하여 dendrogram으로 보았을 때, 동일한 section Liseola에 속하는 F. subglutinans와 F. moniliforme의 종간 유사도는 74.3%로 가장 높았으며, section Elegans와 section Liseola의 서로 다른 section 간의 유사도는 45.4%로 비교적 낮았다. F. napiforme와 F. nygamai의 유사도는 64.7%로 동일 section 내의 종간 유사도와 비슷한 수치를 나타내었다. 이들 두 종은 section Liseola에 대하여 55.2%, section Elegans에 대하여 45.4%의 유사도를 보여 분류학적 위치에 있어서 section Liseola에 더 가까운 경향을 나타내지만, 이는 다른 section 간의 유사도와 비슷한 수치로 하나의 독립된 section으로 추정된다. F. graminearum과 다른 6종 사이의 유사도는 28.2%의 낮은 수치를 나타내어 서로의 유사성이 매우 낮다는 것을 보여 주었다.