• Journal of Microbiology and Biotechnology
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• 제8권6호
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• pp.588-594
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• 1998

Lactic acid bacteria were isolated from Kimchi and screened for bacteriocin production. Strain SE1, identified as Lactobacillus curvatus sp., showed the strongest inhibitory activity against Lactobacillus delbrueckii subsp. delbrueckii. The bacteriocin was inactivated by amyloglucosidase, trypsin, or protease K treatment. However, it maintained its activity under heat treatment at $100^{\circ}C$ for 60 min. The production of the bacteriocin had a growth-related mode and decreased around the early-stationary phase. The optimum temperature for the growth of L. curvatus SE1 was $37^{\circ}C$; however, the optimum temperature for bacteriocin production was $30^{\circ}C$. The bacteriocin activity was decreased by treatment with methanol, butanol, acetone, or chloroform, however, it was not affected by treatment with ethanol, iso-propanol, or cyclohexane. The inhibitory activity of bacteriocin was stable over a wide range of pHs (2 to 11). The bacteriocin from L. curvatus SE1 killed the indicator strain by a bactericidal mode of action. The bacteriocin from L. curvatus SE1 was partially purified by ethanol precipitation and ion exchange chromatography. SDS-polyacrylamide gel electrophoresis was used to determine the molecular weight of the bacteriocin by the bacteriocin activity test. The apparent molecular mass of the bacteriocin produced by L. curvatus SE1 was about 14 kDa.

• 생명과학회지
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• 제18권3호
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• pp.350-356
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• 2008

가지 열매로부터 neutral saline에 의한 추출, $(NH_4)_2SO_4$ 침전 및 Sephadex-G100을 이용한 affinity chromatography에 의해 분리한 lectin의 생화학적 특성을 연구하였다. Trypsin을 처리하지 않은 사람, 쥐와 토끼의 적혈구에서는 혈구 응집반응이 일어나지 않았으며, Trypsin을 처리한 사람, 쥐와 토끼의 적혈구 중에서는 쥐의 적혈구에서만 혈구 응집반응이 일어났다. SDS-PAGE에 의해 가지 lectin의 분자량을 확인한 결과, 19.3 kDa의 단일 band 임이 확인되었다. D-glucose를 포함하는 7개의 탄수화물은 100 mM 이하의 농도에서 lectin과 적혈구의 응집을 저해시키지 않았으므로, 탄수화물에 대한 특이성이 나타나지 않았다. 최적 반응온도 범위는 $10-20^{\circ}C$로서, $20-70^{\circ}C$에서 열에 대해 안정하였으며, 안정된 pH 범위는 pH 6.2-7.2로 확인되었다. $Ca^{2+},\;Co^{2+},\;Cu^{2+},\;Fe^{2+},\;Mg^{2+}$ 및 $Mn^{2+}$ 20 mM 이하의 농도에서는 lectin과 적혈구의 응집을 저해시키지 않았으므로, 금속이온에 대한 특이성이 나타나지 않았다

• 한국해양바이오학회지
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• 제10권2호
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• pp.62-72
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• 2018

An active ingredient with hyaluronidase (HAse) inhibitory effect is one of the anti-aging approaches in cosmeceuticals. Here, red sea cucumbers (RSCs), Stichopus japonicus, from Jeju Island were evaluated to examine their HAse inhibitory and antioxidant activity effect. In this study, RSCs were extracted by six enzymatic hydrolysis (Alcalase; Al, Trypsin; Try, Neutrase; Neu, Pepsin; Pep, Alpha-chymotrypsin; Chy and Protamex; Pro). Alcalase hydrolysate (AlH) showed the highest antioxidant capacities for both of oxygen radical absorbance capacity (ORAC) and trolox equivalent antioxidant capacity (TEAC) methods, compared to those of other hydrolysates, at $66.59{\pm}0.78{\mu}M\;TE/mg$ and $135.78{\pm}3.24{\mu}M\;TE/mg$, respectively. Furthermore, AlH performed the highest capacity of HAse inhibitory with $IC_{50}$ value of 3.21 mg/ml. Thus, RSCs hydrolyzed by Al were chosen to determine the cellular antioxidant activity and hyaluronic acid (HA) production effect on Human immortalized keratinocyte cell line (HaCaT). The results showed that AlH improved the cell viabilities and intracellular reactive oxygen species (ROS) induced by 2,2'-Azobis(2-amidinopropane) dihydrochloride (AAPH) were significantly decreased. In addition, AlH increased HA amount by regulating HYAL2 and HAS2 expressions in the HaCaT cells. Taken together, AlH of RSCs collected from Jeju Island showed HAse inhibitory and antioxidant activities against skin-aging which shows its potentials can be an optional natural bioactive ingredient for novel cosmeceuticals.

• 한국식품영양과학회지
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• 제43권11호
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• pp.1701-1708
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• 2014

참취의 활용 및 생리활성을 증가시킬 수 있는 적정 추출방법을 알아보고자 상온교반, 환류냉각, 가압가열, 저온고압 및 초음파 추출법을 이용하여 추출한 두벌 째 수확 참취 추출물의 생리활성을 비교하였다. 추출 수율은 환류냉각추출, 초음파추출, 가압가열추출, 상온추출, 저온고압추출 순으로 높았다. 폴리페놀, 플라보노이드 함량은 저온고압추출에서 유의적으로 높았고, 상압가열, 고온고압, 초음파추출에서는 대등한 함량을 나타내었다. 페놀화합물 정량의 경우 chlorogenic acid 함량은 가압가열추출에서 유의적으로 높았으며 cynarin 함량의 경우 환류냉각, 저온고압 및 초음파 추출에서 유의적으로 높았고, astragalin 함량의 경우 저온고압추출에서 높은 함량이 검출되었다. Xanthine oxidase(XO), angiotensin I-converting enzyme(ACE), HMG-CoA reductase 저해활성의 경우 저온고압추출 및 초음파 추출물에서 우수한 활성을 나타내었으며, 소화효소 저해활성(${\alpha}$-amylase, trypsin, lipase)에서도 유사한 경향을 나타내었다. 이러한 결과를 종합해 볼 때 저온고압 및 초음파 추출물에서 소재 활용가치가 높을 것으로 사료되며 천연 항산화제 및 기능성 증진을 위한 소재로 이용 가능할 것으로 판단된다.

• 한국수산과학회지
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• 제48권2호
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• pp.178-186
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• 2015

Protease inhibitors (PI) of trypsin and papain as target proteases from the roe of bastard halibut Paralichthys olivaceus were fractionated out using ammonium sulfate precipitation (A), DEAE 650M anion exchange chromatography (D), and Sephacryl S-300 gel filtration (S). The recovery percentages of the fractions with the strongest inhibitory activity for each fractionation method were 13% for the A4 fraction, 21.2% for the D3 fraction, and 21.3% for the S2 fraction, with specific inhibitory activities of the fractions toward trypsin and casein of 168, 139, and 218 U/mg, respectively, while no inhibition of papain was observed. The $IC_{50}$ for the trypsin-specific substrate $N{\alpha}$-benzoyl-$\small{L}$-arginine-p-nitroanilide (BAPNA) was 0.65, 1.55, 2.26, and 2.85 mg/mL for the A4, S2, A3, and D3 fractions, respectively. These results suggest that chromatographic fractionation methods (D and S) based on the molecular mass and charge of the protein were more effective at fractionating PI than was ammonium sulfate precipitation based on protein solubility, and that the bastard halibut roe extract acts as a serine protease inhibitor. Therefore, the PI fraction from fish roe might be useful for inhibiting proteases in foodstuffs, and could constitute an alternative food-grade inhibitor for the surimi industry.

• 한국수산과학회지
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• 제44권2호
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• pp.118-125
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• 2011

In this study, an angiotensin I-converting enzyme (ACE) inhibitor from squid skin was purified and characterized. Squid (Todarodes pacificus) skin protein isolates were hydrolyzed using six commercial proteases: alcalase, ${\alpha}$-chymotrypsin, neutrase, papain, pepsin, and trypsin. The peptic hydrolysate had the highest ACE inhibitory activity. The ACE inhibitory peptide was purified using Sephadex G-25 column chromatography and reverse phase high-performance liquid chromatography (HPLC) with a $C_{18}$ column. The purified ACE inhibitory peptide was identified and sequenced, and found to consist of seven amino acid residues: Ser-Ala-Gly-Ser-Leu-Val-Pro (657Da). The $IC_{50}$ value of the purified ACE inhibitory peptide was 766.2 ${\mu}M$, and Lineweaver-Burk plots suggested that the purified peptide acts as a noncompetitive ACE inhibitor. These results suggest that the ACE inhibitory peptide purified from the peptic hydrolysate of squid skin may be of benefit in developing antihypertensive drugs and functional foods.

• 한국임상약학회지
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• 제9권1호
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• pp.62-65
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• 1999

Inorganic phosphate (Pi) in rabbit plasma was found to block completely phosphotyrosine phosphatase (PTPase) activity without affecting the alkaline phosphatase (ALPase) activity. Our results provided that (1) PTPase activity and inhibitor are separated after G-25 gel-filtration. (2) This inhibitor is heat stable and trypsin-resistant and it can be removed by dialysis using 3 Kd cut-off tubing. (3) The elution pattern of the inhibitor is identical to that of Pi, and by performing a seperate run with inorganic phosphate. (4) The PTPase activity was recovered following an incubation with $CaCl_2$ (10 mM).

• Journal of Ginseng Research
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• 제13권2호
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• pp.215-222
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• 1989

A macromolecular fraction from fresh ginseng root containing mainly polysaccharide showed mild mitogenic activity in human cord blood lymphocytes. 2) When lymphocytes were transformed by Con A or PHA, this fraction could greatly enhance the activity of these lectin mitogens, thus showing a potentiation effect. 3) Although this macromolerular fraction contains saponin and is susceptible to trypsin digestion, it is probably a peptido-glycan in nature on account of its important carbohydrate content and thermal stability. 4) This fraction could not support cancer cell (EAT, K562) growth : its inhibitory effect on these cells remains to be explored.

• 한국미생물·생명공학회지
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• 제15권2호
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• pp.129-134
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• 1987

Snake venom proteinase에 대한 저해물질을 생성하는 Aspergillus 속 균주 MK-24를 토양으로부터 얻어 그 배양액에서 저해물질을 분리하여 Venom proteinase에 대한 작용양상과 안정성에 대한 조사결과는 다음과 같다. Glucose 2%, NaNO$_3$ 0.3%, $K_2$HPO$_4$ 0.02%, MgSO$_4$ㆍ7$H_2O$ 0.02%, KCl 0.02% 조성의 배지(pH 5.0)를 사용하여 3$0^{\circ}C$에서 7일간 배양하여 얻은 배양액을 acetone 심전 활성탄, methanol 침전으로 무정형의 유효분말을 얻었다. 이 물질은 A. b.b. venom proteinase에 대하여 1/2 배양에서 약 70% 저해율을 나타냈으며, A.b.b. venom proteinase에 대한 저해양상을 혼합형이었으며 enzyme-inhibitor complex를 형성하는데 20분 정도가 걸렸다. 반응액중에 Co$^{++}$, $Zn^{++}$, Cu$^{++}$ 등이 존재하면 저해작용이 완전히 억제되었다. 저해율은 사용한 기지리의 종류에 따라 차이가 났다. 즉 casein을 사용했을 때는 hemoglobin이나 albumin보다 저해율이 높았다. 그리고 본 저해물질은 snake venom proteinase 이외에 trypsin에 고농도에서 약간 저해작용을 나타냈으나 pepsin, $\alpha$-chymotrypsin, papain 등과 탄수화물 가수분해효소 등에는 저해능이 없었고, 혈액응고에 대하여는 1.6 $\mu\textrm{g}$/2$m\ell$ 농도 이상에서는 저해작용을 나타내었다. 본 저해물질은 열이나 pH에 대한 안정성이 컸다. 즉, pH처리에 대해서는 37$^{\circ}C$에서 60분 처리로 산이나 alkali에 대해서 대단히 넓은 범위에 걸쳐서 안정하였으며 $65^{\circ}C$에서는 중성까지는 안정하였으나, pH 8 이상에서는 불안정하였고 열처리에 대해서는 10$0^{\circ}C$에서 2시간 처리했을 때에도 잔존활성도가 약 90%로 매우 안정하였다.

• 한국식품영양과학회지
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• 제27권3호
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• pp.367-375
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• 1998

The physicochemical properties of the $\alpha$-amylase inhibitors from black bean and naked barley is Korea were investigated. Preincubation time for maximum inhibition was 30min and no activity change was seen after that time. Optimum pH of the $\alpha$-amylase inhibitors from the black bean and naked barley was pH 7.0 and the inhibitory activities were stable in the range of pH 6.0~8.0 in both phosphate and Tris-HCI buffer solutions. Both inhibitors maintained more than 50% of activity after incubation for 17 min at 7$0^{\circ}C$. The inhibitors from the black bean and naked barley maintained more than 50% of activities after treatment for 40 min and 30 min with pepsin, and 30 min and 50 min with trypsin, respectively. Both inhibitors functioned via a noncompetitive mechanism and were active against porcine pancreatic and human salivary $\alpha$-amylases. The activities of both inhibitors were linear for the ionic stength ranging from 0 to 0.9. The addition of 70 mM maltose to the reaction mixture caused a maximum increase in the relative activities of both inhibitors, but it did not affect the dissociation of the EI complex. The activities of both inhibitors were significantly enhanced by adding 1mM of K+ or Mg2+.