• Journal of Microbiology and Biotechnology
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• 제27권2호
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• pp.251-261
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• 2017

Initially, we screened 18 Aspergillus sojae-like strains from Aspergillus spp. isolated from meju (Korean traditional fermented soybean brick) according to their morphological characteristics. Because members of Aspergillus section Flavi are often incorrectly identified because of their phylogenetic similarity, we re-identified these strains at the morphological and molecular genetic levels. Fourteen strains were finally identified as A. sojae. The isolates produced protease and ${\alpha}-amylase$ with ranges of 2.66-10.64 and 21.53-106.73 unit/g-initial dry substrate (U/g-IDS), respectively, which were equivalent to those of the koji (starter mold) strains employed to produce Japanese soy sauce. Among the isolates and Japanese koji strains, strains SMF 127 and SMF 131 had the highest leucine aminopeptidase (LAP) activities at 6.00 and 6.06 U/g-IDS, respectively. LAP plays an important role in flavor development because of the production of low-molecular-weight peptides that affect the taste and decrease bitterness. SMF 127 and SMF 131 appeared to be non-aflatoxigenic because of a termination point mutation in aflR and the lack of the polyketide synthase gene found in other A. sojae strains. In addition, SMF 127 and SMF 131 were not cyclopiazonic acid (CPA) producers because of the deletion of maoA, dmaT, and pks/nrps, which are involved in CPA biosynthesis. Therefore, A. sojae strains such as SMF 127 and SMF 131, which have high protease and LAP activities and are free of safety issues, can be considered good starters for soybean fermentations, such as in the production of the Korean fermented soybean products meju, doenjang, and ganjang.

• 한국미생물·생명공학회지
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• 제46권4호
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• pp.326-333
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• 2018

본 연구에서는 성장인자, 효소, 펩타이드 등과 같은 기능성 생체 고분자를 대상으로 열에 대한 안정성 및 피부 투과성을 향상시키는 연구를 수행하였다. 이들은 생체 내에서 세포를 활성화 하거나 촉매 작용을 담당하고 있다. 따라서 화장품 등의 외용제에 적용 시, 그 효능의 우수함이 예상되나 열에 대한 불안정성과 높은 분자량으로 피부 투과성이 낮은 단점이 있다. 이를 극복하기 위해 먼저 열에 대한 안정성을 확보할 수 있는 조성을 탐색하였다. 그 결과, 단분자 구조의 humectant 대비 PEG의 길이가 긴 polymeric humectant를 사용한 경우 열에 대한 안정성이 높아지는 것을 확인 할 수 있었다. 한편 이들의 피부 투과를 촉진시키기 위하여 투과 촉진 펩타이드를 phage library로부터 선별하고자 하였다. 투과 촉진 펩타이드는 성장인자, 효소, 펩타이드의 투과 촉진을 위해 공통적으로 사용할 수 있는 구조이다. 그러나 피부의 투과정도는 물질자체의 특성도 영향을 미칠 수 있으나 제형의 성분에 따라서 영향을 받을 수 있다. 본 연구에서는 성장인자를 안정화 할 수 있는 polymeric humectant 제형을 기반으로 투과 촉진 펩타이드 선별을 수행하였다. 그 결과 대조군 펩타이드 대비 투과촉진이 향상된 결과를 확인했을 뿐만 아니라 PBS를 기반으로 선별된 투과 촉진 펩타이드 보다 polymeric humectant 제형에서는 투과도가 우수한 것을 확인 할 수 있었다. 본 연구의 결과는 기능성 생체고분자의 열 안정성 개선 및 피부 투과도 향상에 기여할 수 있을 것으로 기대된다.

• 한국식품과학회지
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• 제37권5호
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• pp.827-832
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• 2005

본 연구는 B. subtilis JM3 단백분해효소 첨가 멸치액젓에서 분리한 peptide의 기능성(항산화, 항암 및 ACE 저해활성)에 대하여 분석하였으며, 그 결과는 다음과 같다. 아미노실소 및 가수분해도는 단백분해효소 첨가구가 대조구에 비해 높았으며, 대조구, B. subtilis JM3 단백분해효소 2 및 4% 첨가구의 멸치액젓은 gel chromatography 상에서, 각각 5, 6 및 7개의 peak를 나타내었다. ACE 저해, DPPH 라디칼소거 및 항암활성은 대조구의 경우 peak 5에서 각각 34.82, 94.11 및 38.29%, 2% 효소 첨가구는 peak 6에서 각각 43.75, 90.01 및 15.61%로 4% 효소 첨가구는 peak 7에서 각각 26.34, 97.00 및 34.76%이었다. 가수분해시 생성된 peptide는 다양한 생리활성을 가지고 있었으며, 앞으로 생리활성을 갖는 peptide의 구조 분석에 관한 연구 및 그 억제 기작에 대한 더 많은 연구가 필요하다고 본다.

• BMB Reports
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• 제35권2호
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• pp.172-177
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• 2002

We previously reported that cholesteryl ester transfer protein (CETP) inhibitory peptides (designated $P_{28}$ and $P_{10})$ have anti-atherogenic effects in hypercholesterolemic rabbits (Biochim. Biophys. Acta (1998) 1391, 133-144). To further investigate those effects, we studied rabbit plasma that was collected after 30 h of a $P_{28}$ or $P_{10}$ injection. We found that there is a strong correlation between the in vivo CETP inhibition effects and alterations of lipoprotein particle size distribution in rabbit plasma, as determined on an agarose gel electrophoresis and gel filtration column chromatography. In vivo effects of the peptide were observed again in C57BL/6 mice that expressed simian CETP. The $P_{28}$ or $P_{10}$ peptide ($7\;{\mu}g/g$ of body weight) that was dissolved in saline was injected subcutaneously into the mice. The $P_{28}$ injection caused the partial inhibition of plasma CETP activity up to 50%, decreasing the total plasma cholesterol concentration by 30%, and increasing the ratio of HD/total-cholesterol concentration by 150% in the CETP-transgenic (tg) mice. The CETP inhibition by the $P_{28}$ or $P_{10}$ made alterations that modulated the size re-distribution of the lipoproteins in the blood stream. Particle size of the very low (VLDL) and low density lipoproteins (LDL) from the peptide-injected group was highly decreased compared to the saline-injected group (determined on the gel filtration column chromatography). In contrast, The HDL particle size of the $P_{28}$-injected group increased compared to the control group (saline-injected). The expression level of the CETP mRNA of the $P_{28}$-injected CETP-tg mouse appeared lower than the saline-injected CETP-tg mouse. These results suggest that the injection of the CETP inhibitory peptide could affect the CETP expression level in the liver by influencing lipoprotein metabolism.

• 한국축산식품학회지
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• 제23권1호
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• pp.63-69
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• 2003

본 연구는 단백질분해효소로 whey protein을 가수분해하여 얻은 가수분해물의 용해도와 유화특성의 변화를 측정하기 위해 실시하였다. Whey protein concentrates를 porcine trypsin(E : S=1 ; 3,000)으로 pH 8.0, 37$^{\circ}C$에서 6시간 동안 가수분해한 whey protein 가수분해물의 유화활성은 분해 4시간째에 가장 높게 나타났으며, 이 때 가수분해도는 5.50%이었다. whey protein의 효소가수분해로 whey protein 중의 $\alpha$-lactalbumin은 분해가 잘 일어나지 않으나 $\beta$-lactoglobulin은 분해 초기부터 급속히 분해되며 유화력 상승에 관여하는 여 러개의 저분자량 peptide를 생성하였다. 가수분해물의 용해도는 가수분해시간이 지남에 따라 증가세를 보이다가 5시간부터 조금씩 감소 추세를 보였으며, pH에 따라서는 등전점 부근인 pH4~5에서 용해도가 가장 낮았으나 가수분해시간이 증가함에 따라 이 부근의 용해도가 현저히 증가하였으며 pH 6이상에서는 pH가 증가함에 따라 용해도도 증가하였다. 유화활성은 용해도의 결과와 거의 비슷한 결과를 나타내었다. 유화 안정성은 분해시간이 지남에 따라 조금씩 증가함을 보여주었으나, 가수분해 4시간부터 pH 8 이상의 PH에서 급격한 증자를 나타내었다.

• 한국식품과학회지
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• 제41권2호
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• pp.179-185
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• 2009

Soybean grit를 이용하여 B. subtilis HA에 의한 고체발효를 통해 얻어진 발효물의 발효 시간에 따른 생리활성물질의 변화 및 항산화능을 평가하였다. 발효시간이 증가함에 따라 혈전분해효소 활성 및 protease 활성은 계속적으로 증가하는 경향을 보였으며, ${\alpha}$-amylase 활성은 발효 5일에서 최대값을 나타내었다. Tyrosine 함량은 발효 5일에서 최대값을 보였으며, 콩 가수분해물중에서 분자량 3,000 Da 이하의 분자량을 갖는 콩 펩타이드 및 갈변화색소는 발효시간이 증가함에 따라서 증가하는 경향을 보였다. Soybean grit 발효물의 총 폴리페놀함량은 주정 추출물이 물 추출물보다 전반적으로 높게 나타났으며 주정 추출물은 발효시간 3일째 18.10 mg/g으로 가장 높은 함량을 나타내었다. 항산화능을 나타내는 DPPH 라디칼 소거능은 발효시간 3일째 가장 높은 값을 보였으며, ABTS 라디칼 소거능은 발효 기간이 증가함에 따라 대조군에 비해 증가되었다.

• 한국축산식품학회지
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• 제34권6호
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• pp.844-851
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• 2014

This study focused on the anti-oxidative and collagenase- and elastase inhibition effects of low molecular weight peptides (LMP) from commercial Jeju horse leg bone hydrolysates (JHLB) on pancreatin, via enzymatic hydrolysis. Cell viability of dermal fibroblasts exposed to UVB radiation upon treatment with LMP from JHLB was evaluated. Determination of the antioxidant activity of various concentrations of LMP from JHLB were carried out by assessing 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2-azino-bis-3-ethybenzothiazoline-6-sulphonic acid (ABTS) radical scavenging activity, ferric reducing antioxidant power (FRAP), and oxygen radical absorbance capacity (ORAC). The DPPH radical scavenging activity of LMP from JHLB (20 mg/mL) was 92.21% and ABTS radical scavenging activity (15 mg/mL) was 99.50%. FRAP activity (30 mg/mL) was $364.72{\mu}M/TE$ and ORAC activity (1 mg/mL) was $101.85{\mu}M/TE$. The anti-wrinkle potential was assessed by evaluating the elastase- and collagenase inhibition potential of these LMP. We found that 200 mg/mL of LMP from JHLB inhibited elastase activity by 41.32%, and 100 mg/mL of LMP from JHLB inhibited collagenase activity by 91.32%. The cell viability of untreated HS68 human dermal fibroblasts was 45% when exposed to a UVB radiation dose of $100mJ/cm^2$. After 24 h of incubation with $500{\mu}g/mL$ LMP from JHLB, the cell viability increased to 60%. These results indicate that LMP from JHLB has potential utility as an anti-oxidant and anti-wrinkle agent in the food and cosmetic industry. Additional in vivo tests should be carried out to further characterize these potential benefits.

• 한국축산식품학회지
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• 제42권3호
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• pp.441-454
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• 2022

This study aimed to determine the effect of enzyme, guar gum, and pressure processing on the digestibility and physicochemical properties of age-friendly liver sausages. Liver sausages were manufactured by adding proteolytic enzyme (Bromelain) and guar gum, and pressure-cooking (0.06 MPa), with the following treatments: control, without proteolytic enzyme; T1, proteolytic enzyme; T2, proteolytic enzyme and guar gum; T3, pressure-cooking; T4, proteolytic enzyme and pressure-cooking; T5, proteolytic enzyme, guar gum, and pressure-cooking. The pH was high in the enzyme- and pressure-processed groups. The pressure-processed groups had lower apparent viscosity than other cooking groups, and it decreased during enzyme treatment. Hardness was lower in the enzyme- and pressure-processed groups than in the control, and the T4 was the lowest. Digestibility was the highest in T4 at 82.58%, and there was no significant difference with that in T5. The general cooking group with enzyme and guar gum also showed higher digestibility than the control (77.50%). As a result of the sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the enzyme- and pressure-treated groups (T4, T5) were degraded more into low-molecular-weight peptides (≤37 kDa) than the control and other treatments. Viscoelasticity showed similar trends for viscous and elastic moduli. Similarly, combined pressure processing and enzymatic treatment decreased viscoelasticity, while guar gum increased elasticity but decreased viscosity. Therefore, the tenderized physical properties and improved digestibility by enzyme and pressurization treatment could be used to produce age-friendly spreadable liver sausages.

• Macromolecular Research
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• 제13권3호
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• pp.167-175
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• 2005

This review explores recent works involving the use of the self-assembled nanoparticles of bile acid-modified glycol chitosans (BGCs) as a new drug carrier for cancer therapy. BGC nanoparticles were produced by chemically grafting different bile acids through the use of l-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC). The precise control of the size, structure, and hydrophobicity of the various BGC nanoparticles could be achieved by grafting different amounts of bile acids. The BGC nanoparticles so produced formed nanoparticles ranging in size from 210 to 850 nm in phosphate-buffered saline (PBS, pH=7.4), which exhibited substantially lower critical aggregation concentrations (0.038-0.260 mg/mL) than those of other low-molecular-weight surfactants, indicating that they possess high thermodynamic stability. The SOC nanoparticles could encapsulate small molecular peptides and hydrophobic anticancer drugs with a high loading efficiency and release them in a sustained manner. This review also highlights the biodistribution of the BGC nanoparticles, in order to demonstrate their accumulation in the tumor tissue, by utilizing the enhanced permeability and retention (EPR) effect. The different approaches used to optimize the delivery of drugs to treat cancer are also described in the last section.

• Asian Pacific Journal of Cancer Prevention
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• 제16권16호
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• pp.7039-7044
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• 2015

Background: Many functional molecules controlling diverse cellular function are included in low-molecular weight proteins and peptides. Materials and Methods: To identify proteins controlling function in lung adenocarcinomas (AC), we performed two-dimensional gel electrophoresis employing tricine-SDS polyacrylamide in the second dimension (tricine 2-DE). This system was able to detect proteins under 1 kDa even with post-translational modifications. To confirm the utility of detected proteins as novel tumor markers for AC, we performed immunohistochemical analysis using 170 formalin-fixed and paraffin-embedded lung AC tissues. Results: Tricine 2-DE revealed that five proteins including S100A16 were overexpressed in lung AC-derived cells compared with lung squamous cell carcinoma, small cell carcinoma, and large cell neuroendocrine carcinoma-derived cells. Immunohistochemically, S100A16 showed various subcellular localization in lung cancer tissues and a membranous staining status was correlated with the T-factor (P=0.0008), pathological stage (P=0.0015), differentiation extent (P=0.0001), lymphatic invasion (P=0.0007), vascular invasion (P=0.0001), pleural invasion (P=0.0087), and gender (P=0.039), but not with the age or smoking history. More importantly, membranous staining of S100A16 was significantly correlated with a poorer overall survival of either stage I (P=0.0088) or stage II / III (P=0.0003) lung AC patients, and multivariate analysis confirmed that membranous expression of S100A16 was an independent adverse prognostic indicator (P=0.0001). Conclusions: The present results suggest that S100A16 protein is a novel prognostic marker for lung AC.