• Food Science and Biotechnology
• /
• 제18권2호
• /
• pp.317-322
• /
• 2009

The proteins from 15 types of cultivar of hot peppers cultivated in Korea were extracted and its allergenicity was investigated by immunoblotting and enzyme-linked immunosorbent assay (ELISA). The immunoblotting of hot pepper proteins extracts (HPEs) against serum of hot pepper sensitized patients revealed dominant IgE binding to 14, 37, and 40 kDa molecules. The specific levels of IgE to HPEs sample No. 1, 3, and 7 were much higher than the other samples in patients. Also, IgE binding capacity of HPEs were not reduced by thermal processing and digestion in ELISA using human IgE antibody acquired from hot pepper sensitized patients. By means of Western blotting using anti-thaumatin IgY, thaumatin-like protein (TLP) acting as allergen in several plants and fruits was detected in tested hot peppers. This study demonstrates that the antigenic protein in hot peppers are present but are differently contained according to cultivars.

• 대한한의학회지
• /
• 제31권2호
• /
• pp.137-148
• /
• 2010

Objective: This study aimed to evaluate the protective effects of Gamipalmi-hwan (GPH) on elastase-induced lung cell injury. Materials and Methods: As an in vitro model of emphysema, the current study was performed to investigate potential activity of GPH in regulating injury responses of A549 human type II cell line mediated by elastase treatment. Results: GPH treatment increased the number of A549 cells which was reduced by elastase digestion. Elastin protein level, which was reduced by elastase treatment, was increased by GPH treatment. Labeling intensity with caspase 3 protein in elastase-treated cells was reduced by GPH treatment. Both Erk1/2 and Cdc2 protein levels, which were decreased by elastase treatment, were increased to a level similar to that of the normal cells. mRNA levels encoding IL-$1{\beta}$ and TNF-$\alpha$ were increased by elastase and then down-regulated by GPH. Conclusion: The present data suggest that A549 cells are subjected to inflammatory damage by elastase and can be recovered by GPH treatment. Further studies examining the protective activity of GPH in elastase-treated lung tissue would be useful for therapeutic strategies of emphysema treatment.

• Bulletin of the Korean Chemical Society
• /
• 제16권5호
• /
• pp.401-406
• /
• 1995

The P. fragi lipase overexpressed in E. coli as a fusion protein of 57 kilodalton (kDa) has been purified through glutathione-agarose affinity chromatography by elution with free glutathione. The general properties of the purified GST-fusion protein were characterized by observing absorbance of released p-nitrophenoxide at 400 nm which was hydrolyzed from the substrate p-nitrophenyl palmitate. The optimum condition was observed at 25 $^{\circ}C$, pH 7.8 with 0.4 ${\mu}g$ of protein and 1.0 mM substrate in 0.6% (v/v) TritonX-100 solution. Also the lipase was activated by Ca+2, Mg+2, Ba+2 and Na+ but it was inhibited by Co+2 and Ni+2. pGEX-2T containing P. fragi lipase gene as expression vector was named pGL191 and used as a template for the site-directed mutagenesis by sequential PCR steps. A Ser-His-Asp catalytic triad similar to that present in serine proteases may be present in Pseudomonas lipase. Therefore, the PCR fragments replacing Asp217 to Arg and His260 to Arg were synthesized, and substituted for original fragment in pGL19. The ligated products were transformed into E. coli NM522, and pGEX-2T harboring mutant lipase genes were screened through digestion with XbaI and StuI sites created by mutagenic primers, respectively. No activity of mutant lipases was observed on the plate containing tributyrin. The purified mutant lipases were not activated on the substrate and affected at pH variation. These results demonstrate that Asp217 and His260 are involved in the catalytic site of Pseudomonas lipase.

• 한국축산식품학회지
• /
• 제44권4호
• /
• pp.951-965
• /
• 2024

Lactiplantibacillus plantarum is a valuable potential probiotic species with various proven health-beneficial effects. L. plantarum LM1001 strain was selected among ten strains of L. plantarum based on proteolytic activity on whey proteins. L. plantarum LM1001 produced higher concentrations of total free amino acids and branched-chain amino acids (Ile, Leu, and Val) than other L. plantarum strains. Treatment of C2C12 myotubes with whey protein culture supernatant (1%, 2% and 3%, v/v) using L. plantarum LM1001 significantly increased the expression of myogenic regulatory factors, such as Myf-5, MyoD, and myogenin, reflecting the promotion of myotubes formation (p<0.05). L. plantarum LM1001 displayed β-galactosidase activity but did not produce harmful β-glucuronidase. Thus, the intake of whey protein together with L. plantarum LM1001 has the potential to aid protein digestion and utilization.

• 한국식품영양과학회지
• /
• 제41권9호
• /
• pp.1310-1314
• /
• 2012

본 실험은 제초제 내성을 가지는 CP4EPSPS를 도입시킨 유전자재조합 콩을 이용하여 인공위액의 비율에 따른 도입 단백질의 소화특성을 평가하고, 열처리가 유전자재조합 콩의 도입단백질의 소화성에 미치는 영향을 살펴보았다. 유전자재조합 콩과 펩신의 비율에 따른 도입단백질 CP4EPSPS의 소화성은 인공위액 즉 펩신의 양에 따라 차이를 나타내 유전자재조합 식품의 도입단백질에 대한 소화성 평가에서 펩신의 양이 중요한 요인이 됨을 알 수 있었다. 또한 유전자 재조합 콩에 대한 예열처리가 도입단백질 CP4EPSPS의 펩신에 대한 내성을 많이 감소시켜 소화성을 높이는 것으로 나타났다. 따라서 앞으로 유전자재조합식품의 안전성을 평가하는 in vitro 소화평가 시험에서 생리학적 측면에 근거한 인공위액의 비율을 반영함으로써 보다 정확한 도입단백질의 소화성에 대한 평가가 필요하리라 사료되며, 또한 콩의 주요 섭취방법인 열처리에 따른 도입단백질의 소화특성을 평가함으로써 유전자재조합 콩의 안전성에 대한 신뢰도 제고를 위한 기초자료가 될 것으로 기대된다.

• 생명과학회지
• /
• 제19권11호
• /
• pp.1591-1597
• /
• 2009

이 실험의 목적은 해조류 중 특히 필수 아미노산 함량이 높은 매생이 열수 추출물(Capsosiphon fulvescens Extracts, CFE) 5%를 첨가한 식이를 급이 시켜 흰쥐에게 10일간의 단백질 소화율, 혈중 필수 아미노산 변화에 미치는 영향을 규명하고 사료의 품질 및 필수 아미노산 대사와 관련된 연구에 기초자료를 마련하는데 그 목적을 두고 본 실험을 하였다. CFE의 급이에 따른 단백질 소화율을 살펴 본 결과, casein 그룹의 경우 단백질 소화율이 약 97%인 반면 CFE가 포함된 그룹의 경우 87%로 유의적으로 감소하는 것으로 나타났다. 실험 식이에 CFE 첨가 시 단백질 소화율이 유의적으로 감소시키는 것으로 나타났다. 간문맥 혈청 중의 아미노산에서 특히 필수 아미노산의 혈중 흡수에 미치는 영향을 식이 후 30, 60, 90, 120분별로 살펴보았다. 총아미노산과 필수 아미노산의 농도는 두개의 군에서 차이가 없었다. BCAA에 속하는 Leu, Ile, Val은 C군에 비해 CFE군에서 흡수속도가 증가하였으며 AAA에 속하는 Phe 또한 동일한 경향을 보여주었다. 간문맥 혈중의 Lys, Thr, Met의 경우, C군에 비해 CFE군에서 흡수속도가 지연되는 것으로 나타났다. 이 실험을 통해 식이섬유인 매생이 열수추출물을 섭취함으로서 간이나 뇌, 조직 등으로 흡수되는 혈중 필수 아미노산의 흡수속도를 증가시키거나 지연시킴으로서 단백질 대사에 영향을 미치는 것으로 보인다. 따라서 단백질 식이에 매생이 열수추출물을 첨가할 경우 혈중 아미노산 흡수정도에 어떠한 영향을 미치는지에 대한 기초자료를 마련 하였으며 이후 더 많은 연구가 필요한 것으로 보인다.

• 한국식품영양과학회지
• /
• 제13권1호
• /
• pp.97-106
• /
• 1984

남대양(南大洋)에 다량 서식하고 있는 중요한 단백질자원(蛋白質資源)인 크릴을 보다 유효하게 식량(食糧)으로 이용하기 위한 방안의 하나로 크릴을 원료로 하여 그 자체에 존재하는 활성이 높은 자가소화효소(自家消化酵素)나 단백질분해효소(蛋白質分解酵素)로써 크릴간장 제조(製造)를 시도(試圖)하였다. 마쇄한 크릴에 동량(同量)의 물을 첨가하여 가수분해(加水分解)시킬 때의 최적가수분해조건(最適加水分解條件) 및 저장안정성(貯藏安定性)을 검토(檢討)하고 아울려 제품의 정미성분(呈味成分)을 분석(分析)하였다. 자가소화(自家消化)에 의한 경우와 bromelain, complex enzyme을 첨가한 시료(試料) 모두 $52.5^{\circ}C$에서 최대활성을 나타내었고, 분해시간(分解時間)은 3시간(時間)이 적합하였으며, 효소농도(醴素濃度)는 bromelain의 경우 0.5 %, complex enzyme은 5%가 가장 좋았다. 그리고 pH는 자가소화(自家消化)나 complex enzyme을 첨가하여 분해(分解)하였을 경우는 7.0${\sim}$7.5, bromelain을 첨가하여 분해(分解)하였을 때는 6.5부근에서 가장 활성이 높았다. 이와 같은 최적가수부해조건(最適加水分解條件)에서 크릴을 자가소화(自家消化)시킨 후 $100^{\circ}C$, 20 분간(分間) 불활성화한 다음 여과한 가수분해물(加水分解物)에 식염(食鹽)(10 %)과 벤조산(0.06%)이나 알코올(3 %)을 첨가하여 멸균한 유리병에 밀봉(貯減)한 결과 $37^{\circ}C$에서 한달간 저장(貯藏)하여도 화학적, 미생물적 및 관능적으로 안정하였다. 또한 관능적인 맛으로 보아 식염(食鹽) 10 % 중 5 % 정도까지는 나트륨염(鹽) 대신 칼륨염(鹽)을 대체 첨가할 수 있었다. 크릴간장의 단백질가수분해율(蛋白質加水分解率)은 자가소화법(自家消火法)인 경우 83.2%, bromelain 첨가구는 89.7%, complex enzyme 첨가구는 92.7 %이었다. 자가소화법(自家消化法), bromelain 또는 complex enzyme첨가에 의해 제조(製造) 된 크릴간장의 유리(遊離)아미노산(酸) 중 함량이 많은 것은 lysine, arginine, leucine, proline, alanine 및 valine으로서 전 유리(遊離)아미노산(酸)에 대해 각각 58.8 %, 56.0 %, 55.3 %를 차지하였다. 그리고 전엑스분망(分望) 소(素)에 대하여 유리(遊離)아미노산(酸)이 차지하는 비율은 각각 67.4 %,69.4%, 69.8 %이었다. 핵산관련물질(核酸關聯物質) 중 함량이 가장 많은 것은 hypoxanthine이었고, 다음이 5’-IMP였다. TMAO, betaine, 총 creatinine은 함량이 적었다. 관능검사결과(官能檢査結果) 자가소화(自家消化)시킨 크릴간장은 효소(酵素)처리한 것이나 재래식 콩간장에 비하여 품질 면에서 손색이 없고 저장성(貯藏性)이 좋은 크릴간장을 제조(製造)할 수 있다는 결론을 얻었다.

• Animal Bioscience
• /
• 제36권8호
• /
• pp.1190-1198
• /
• 2023

Objective: To our knowledge, there are few studies on the correlation between internal structure of fermented products and nutrient delivery from by-products from coffee processing in the ruminant system. The objective of this project was to use advanced mid-infrared vibrational spectroscopic technique (ATR-FT/IR) to reveal interactive correlation between protein internal structure and ruminant-relevant protein and energy metabolic profiles of by-products from coffee processing affected by added-microorganism fermentation duration. Methods: The by-products from coffee processing were fermented using commercial fermentation product, called Saus Burger Pakan, consisting of various microorganisms: cellulolytic, lactic acid, amylolytic, proteolytic, and xylanolytic microbes, for 0, 7, 14, 21, and 28 days. Protein chemical profiles, Cornell Net Carbohydrate and Protein System crude protein and CHO subfractions, and ruminal degradation and intestinal digestion of protein were evaluated. The attenuated total reflectance-Ft/IR (ATR-FTIR) spectroscopy was used to study protein structural features of spectra that were affected by added microorganism fermentation duration. The molecular spectral analyses were carried using OMNIC software. Molecular spectral analysis parameters in fermented and non-fermented by-products from coffee processing included: Amide I area (AIA), Amide II (AIIA) area, Amide I heigh (AIH), Amide II height (AIIH), α-helix height (αH), β-sheet height (βH), AIA to AIIA ratio, AIH to AIIH ratio, and αH to βH ratio. The relationship between protein structure spectral profiles of by-products from coffee processing and protein related metabolic features in ruminant were also investigated. Results: Fermentation decreased rumen degradable protein and increased rumen undegradable protein of by-products from coffee processing (p<0.05), indicating more protein entering from rumen to the small intestine for animal use. The fermentation duration significantly impacted (p<0.05) protein structure spectral features. Fermentation tended to increase (p<0.10) AIA and AIH as well as β-sheet height which all are significantly related to the protein level. Conclusion: Protein structure spectral profiles of by-product form coffee processing could be utilized as potential evaluators to estimate protein related chemical profile and protein metabolic characteristics in ruminant system.

• BMB Reports
• /
• 제38권2호
• /
• pp.225-231
• /
• 2005

Acid-labile subunit (ALS) is a component of the 150-kDa insulin-like growth factor-binding protein-3 (IGFBP-3) complex, which, by sequestering the majority of IGFs-I and -II and thereby prolonging the half-life of them in plasma, serves as a circulating reservoir of IGFs in mammalian species. A pGEX-2T plasmid and a baculovirus expression constructs harboring a coding sequence for glutathione-S transferase (GST)-porcine ALS (pALS) fusion protein were expressed in BL21(DE3) E. coli and Sf9 insect cells, respectively. The expressed protein was purified by glutathione or Ni-NTN affinity chromatography, followed by cleavage of the fusion protein using Factor Xa. In addition, pALS and hIGFBP-3 were also produced in small amounts in the Xenopus oocyte expression system which does not require any purification procedure. A 65-kDa pALS polypeptide was obtained following the prokaryotic expression and the enzymatic digestion, but biochemical characterization of this polypeptide was precluded because of an extremely low expression efficiency. The baculovirus-as well as Xenopus-expressed pALS exhibited the expected molecular mass of 85 kDa which was reduced into 75 and 65 kDa following deglycosylation of Asn-linked carbohydrates by Endo-F glycosidase, indicating that the expressed pALS was properly glycosylated. Moreover, irrespective of the source of pALS, the recombinant pALS and hIGFBP-3 formed a 130-kDa binary complex which could be immunoprecipitated by anti-hIGFBP-3 antibodies. Collectively, results indicate that an authentic pALS protein can be produced by the current expression systems.

• Asian-Australasian Journal of Animal Sciences
• /
• 제17권8호
• /
• pp.1062-1068
• /
• 2004

Leptin has a major role in the regulation of food intake and energy homeostasis. In addition, leptin participates in many physiological functions including regulation of lipid metabolism. Bovine recombinant leptin protein was produced in E. coli cells in order to understand function of leptin in the regulation of lipid metabolism. The leptin expression vector was constructed in pGEX-4T-3 vector and transformed into E. coli BL21 cells. Expression of the GST-leptin fusion protein was induced with IPTG. The fusion protein was purified using glutathione sepharose 4B batch method, and the recombinant leptin was eluted after thrombin protease digestion. The effect of leptin on glucose transport was examined in the differentiated adipocytes of 3T3-L1 cells. Leptin had no effect on basal and insulin-stimulated glucose transport in 3T3-L1 cells (p>0.05). Effect of recombinant leptin on glucose and acetate transport was examined in adipose tissues of Korean cattle (Hanwoo). Insulin stimulated glucose transport in both intramuscular and subcutaneous adipose tissues (p<0.05), but leptin did not affect glucose transport in both adipose tissues (p>0.05). Insulin stimulated acetate transport in bovine adipose tissues (p<0.05), but leptin did not affect acetate transport (p>0.05). Northern and RT-PCR analyses showed that mRNA levels of uncoupling protein-2 were increased by leptin treatment in 3T3-L1 cells without statistical difference (p>0.05). In conclusion, bovine recombinant leptin did not affect glucose and acetate transport in both 3T3-L1 adipocytes and bovine adipose tissues, while it stimulates UCP-2 mRNA expression in 3T3-L1 cells.