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한국미생물생명공학회> 한국미생물·생명공학회지> 아스파라거스를 이용한 전통장류의 항산화 효과

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아스파라거스를 이용한 전통장류의 항산화 효과

Antioxidant Activity of Korea Traditional Fermented Sauces Made with Asparagus

김예은 ( Yeeun Kim ) , 지근호 ( Keunho Ji )
  • : 한국미생물생명공학회
  • : 한국미생물·생명공학회지 49권1호
  • : 연속간행물
  • : 2021년 03월
  • : 95-100(6pages)
한국미생물·생명공학회지

DOI


목차

서 론
재료 및 방법
결과 및 고찰
요 약
Conflict of Interest
References

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초록 보기

본 연구에서는 아스파라거스가 함유된 전통장류의 항산화 활성을 확인하였다. 아스파라거스의 항산화능은 농도와 무관하게 나타났으며 70 mg/ml에서 가장 높은 활성을 보였다. 아스파라거스가 함유된 메주의 경우 유기용매 분획별로 항산화 활성을 측정하였으며, 분획 중 물층과 에틸아세테이트 층에서 높은 항산화 활성이 확인되었다. 총 폴리페놀 함량 측정 결과 물층의 경우 tannic acid 1.437 mg/g, 에틸아세테이트 층의 경우 tannic acid 0.77 mg/g과 유사한 활성을 보였다. DPPH 라디칼 소거능과 ABTS 라디칼 소거능으로 측정한 항산화 활성은 물층의 경우 각각 27.6%, 9.04%의 활성을 보였으며, 에틸아세테이트 층의 경우 각각 10.7%, 52.4%의 활성을 보였다. 아스파라거스 함유 전통장류의 항산화 활성은 95% 에탄올 추출물을 이용하여 측정하였다. 가장 높은 활성을 보인 장류는 간장이며, 간장은 가장 넓은 범위의 농도에서도 좋은 활성을 나타내었다. 본 연구를 통하여 아스파라거스가 함유된 전통 장류의 항산화능이 증가하는 것을 확인하였다. 추후 진행될 연구를 통해 전통장류의 우수성이 증대되고, 새로운 장류의 개발에 기여하여 전통장류의 세계화 및 가치를 높일 수 있을 것으로 기대된다.
Asparagus have anti-cancer, antioxidant, and anti-inflammatory properties. Since the high functionality of Korean traditional sauces was discovered, various studies have been conducted using asparagus. In this study, the antioxidant activity of traditional sauces containing asparagus was confirmed. The antioxidant capacity of asparagus was independent of the concentration and showed the highest activity at 70 mg/ml. For instance, concerning Meju containing asparagus, high antioxidant activity was observed in water and ethyl acetate extracts; the total polyphenol content was equivalent to 1.4 mg/g and 0.7 mg/g of tannic acid in the water and ethyl acetate fractions respectively. The DPPH and ABTS radical scavenging activities were 27.6% and 9.04%, respectively, in the water layer, and 10.7% and 52.4%, respectively, in the ethyl acetate layer. Additionally, the antioxidant activity of asparagus-containing traditional sauces was measured using 95% ethanol extracts; soy sauce showed the highest activity, retained in a wide range of concentrations.

UCI(KEPA)

I410-ECN-0102-2022-400-000428868

간행물정보

  • : 자연과학분야  > 생물
  • : KCI등재
  • : SCOPUS
  • : 계간
  • : 1598-642x
  • : 2234-7305
  • : 학술지
  • : 연속간행물
  • : 2002-2022
  • : 1249


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1Properties of a Novel Glutamate Decarboxylase (GAD) from Levilactobacillus brevis B737 Isolated from Cabbage Kimchi

저자 : Tae Jin Kim , Min Jae Kim , Bong Sin Kim , Ji Yeon Yoo , Yun Ji Kang , Jeong Hwan Kim

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 50권 3호 발행 연도 : 2022 페이지 : pp. 319-327 (9 pages)

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γ-Aminobutyric acid (GABA) is a multi-functional compound with broad applications for food industry. GABA producing bacteria were isolated from cabbage kimchi. Among them, B737 was the best GABA producer when culture supernatants were analyzed by TLC. B737 was identified as Levilactobacillus brevis by 16S rRNA gene sequencing. Its glutamate decarboxylase (GAD) gene was cloned by PCR and the nucleotide sequence determined. B737 GAD consisting of 485 amino acids is the largest in size among GADs reported from LAB so far. gadB from L. brevis B737 was overexpressed in Escherichia. coli BL21(DE3) using pET26b(+). The recombinant GAD was purified and its size was 55 kDa by SDS-PAGE. Maximum GAD activity was observed at pH 5 and 40℃ and the activity was dependent on pyridoxal 5'-phosphate. Km and Vmax of recombinant GAD were 6.2 ± 0.06 mM and 0.34 ± 0.002 mM/min, respectively. L. brevis B737 can be used as a starter for fermented foods with high GABA contents.

KCI등재 SCOPUS

2Antibiofilm Activity and Binding Specificity of Polyclonal DNA Aptamers on Staphylococcus aureus and Escherichia coli

저자 : Arizah Kusumawati , Apon Zaenal Mustopa , Rifqiyah Nur Umami , Adi Santoso , I Wayan Teguh Wibawan , Agus Setiyono , Mirnawati Bachrum Sudarwanto

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 50권 3호 발행 연도 : 2022 페이지 : pp. 328-336 (9 pages)

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Aptamers are short, chemically synthesized, single-stranded DNA or RNA oligonucleotides that fold into unique three-dimensional structures. In this study, we aim to determine the antibiofilm activity and binding specificity of the six polyclonal DNA aptamers (S15K3, S15K4, S15K6, S15K13, S15K15, and S15K20) on Staphylococcus aureus BPA-12 and Escherichia coli EPEC 4. Aptamer S15K6 showed the highest percentage of antibiofilm activity against S. aureus BPA-12 (37.4%) as shown by the lowest OD570 value of 0.313. Aptamer S15K20 showed the highest percentage of antibiofilm activity against E. coli EPEC 4 (15.4%) as shown by the lowest OD570 value of 0.515. Aptamers S15K13 and S15K20 showed antibiofilm activities against both S. aureus BPA-12 and E. coli EPEC4, and thus potentially have broad reactivity. Furthermore, based on the binding capacity and Kd values from our previous study, the binding specificity assay of selected polyclonal DNA aptamers (S15K3 and S15K15) against S. aureus BPA-12, E. coli EPEC 4, S. aureus BPA-6, S. agalactiae, E. coli MHA-6, and Listeria monocytogenes were performed using qPCR. Aptamers S15K3 and S15K15 showed specific binding to S. aureus BPA-12, E. coli EPEC 4, S. aureus BPA-6, and S. agalactiae, but could not bind to E. coli MHA-6 and L. monocytogenes. Therefore, this study showed that the polyclonal DNA aptamers have antibiofilm activity and were able to bind to S. aureus BPA-12 and E. coli EPEC 4 bacteria.

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3LPS에 의해 활성화된 대식세포에서 애플망고 껍질(Mangifera indica L. Peel)의 항염증 효능 검증

저자 : 김효민 ( Hyo-min Kim ) , 유단희 ( Dan-hee Yoo ) , 이인철 ( In-chul Lee )

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 50권 3호 발행 연도 : 2022 페이지 : pp. 337-346 (10 pages)

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최근 국내 기후 변화로 인해 열대 과일 재배가 가능해지게 되었다. 특히 대표적으로 제주도에서 애플망고의 재배가 활발히 이루어지고 있으며 이로 인해 소비자들의 수요가 증가하고 있다. 그러나 애플망고 과육을 제외한 껍질과 같은 부산물은 활용도가 떨어져 폐기되고 있는 실정이다. 이에 본 연구에서는 부산물로서 폐기되어지는 애플망고 껍질을 활용하기 위해 이를 이용하여 항산화 및 항염증 효과를 검증하고자 열수 및 70% 에탄올로 추출하여 실험을 진행하였다. 먼저 애플망고 껍질의 열수 및 70% 에탄올 추출물의 총 폴리페놀 함량을 확인하였으며, 그 결과 66.08 ± 0.12 mg TAE/100 g, 100.13 ± 0.23 mg TAE/100 g의 함량을 나타내었다. 항산화 활성을 측정하기 위해 전자공여능과 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) 라디칼 소거능을 측정하였다. 전자공여능 측정결과, 1,000 μg/ml 농도에서 열수 추출물은 86.89%가 나타났고, 70% 에탄올 추출물은 94.06%를 확인하였다. ABTS 라디칼 소거능을 측정 결과로 1,000 μg/ml 농도에서 열수 추출물은 80.62%를 나타내었고, 70% 에탄올 추출물은 98.66%의 소거능을 확인하였다. 이로 인해 애플망고 껍질 추출물의 항산화 활성은 대조군인 비타민 C와 유사한 효과를 가진 것을 확인하였다. 애플망고 껍질 추출물의 세포 생존율을 측정한 결과, 대식세포인 RAW 264.7 세포에서는 500 μg/ml 농도에서 99% 이상의 세포 생존율을 확인하였다. 이후 세포실험은 세포생존율이 99% 이상인 농도구간을 설정하여 진행하였다. 염증성 매개 물질로 알려진 nitric oxide (NO)에 대한 생성 억제 효과를 확인한 결과, 애플망고 껍질의 열수 추출물은 46%의 억제 효과가 나타났고, 70% 에탄올 추출물은 40%의 억제 효과를 확인하여 애플망고 추출물의 NO 생성량 저해 효과를 가진 것을 확인하였다. 염증 매개 인자의 억제 효과를 확인하기 위해 RAW 264.7 세포를 이용하여 iNOS와 COX-2의 단백질 및 mRNA 발현량을 측정하였다. 그 결과, LPS 단독 처리군에 비해 애플망고 껍질 추출물을 처리하였을 때 iNOS와 COX-2의 단백질 발현량은 최종 농도인 500 μg/ml에서 열수 추출물은 12%, 97% 억제하는 것을 확인하였고, 70% 에탄올 추출물은 62%, 91% 억제하는 것을 확인하였다. 또한 LPS 단독 처리군에 비해 iNOS와 COX-2의 mRNA 발현량 결과, 500 μg/ml 농도에서 열수 추출물은 3%, 33%의 억제 효과를 확인하였고, 70% 에탄올 추출물은 29%, 36%의 억제 효과가 나타나는 것을 확인하였다. 본 연구 결과들을 토대로 애플망고 껍질이 항산화 및 항염증 활성을 갖고 있는 것을 검증하여 천연 소재로서의 활용 가치가 높을 것으로 사료된다.


The purpose of this study was to investigate the antioxidant and anti-inflammatory activities of hot water (AMPW) and 70% ethanol (AMPE) extracts of apple mango (Mangifera indica L.) peel. The antioxidant activities were measured using a total polyphenol, electron-donating, 2,2'-azinobis [3-ethylbenzothiazoline-6-sulfonic acid] (ABTS) radical scavenging assay. The total polyphenol content of AMPW and AMPE was 66.08 ± 0.62 mg TAE/100 g and 100.13 ± 0.23 mg TAE/100 g, respectively. As a result of measuring the electron-donating ability, at a concentration of 1,000 μg/ml, AMPW and AMPE showed an effectiveness of 86% and 94%, respectively. The ABTS assay showed 80% and 98% respective radical scavenging activity for AMPW and AMPE, at a concentration of 1,000 μg/ml. The cell viability on macrophage cells was performed using a 3-[4,5-dimethyl-thiazol-2-yl]-2,5-diphenyl-tetrazoliumbromide (MTT) assay, and the results showed more than 90% cell viability at a 100 μg/ml concentration. Anti-inflammatory activity was verified by confirming nitric oxide (NO) production inhibitory activity, inducible nitric oxide synthase (iNOS), and cyclooxygenase- 2 (COX-2) protein and mRNA expression inhibitory activity from lipopolysaccharide (LPS)-treated RAW 264.7 cells. The NO production inhibitory effects were measured using the Griess assay, which confirmed 45% and 40% inhibition after treatment with AMPW and AMPE, respectively. Moreover, the protein and mRNA expression of inflammatory-related factors iNOS and COX-2, decreased in a concentration-dependent manner. In conclusion, this study showed antioxidant and anti-inflammatory effects of Mangifera indica L. peel and revealed its promising potential for application as an antioxidant and anti-inflammatory agent.

KCI등재 SCOPUS

4Synbiotics 첨가 사료 투여가 넙치(Paralichthys olivaceus)의 성장과 비특이적 면역 반응 및 세균성 감염 폐사에 미치는 영향

저자 : 문채윤 ( Chae-yun Moon ) , 손홍주 ( Hong-ju Son ) , 허문수 ( Moon-soo Heo )

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 50권 3호 발행 연도 : 2022 페이지 : pp. 347-353 (7 pages)

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본 연구는 기존의 probiotics로서의 안전성을 입증한 B. cabrialesii sp. K1과 prebiotics 중 inulin 3%와 혼합하여 synbiotics로서의 넙치의 성장과 비특이적 면역 반응 및 세균성 감염에 따른 폐사에 미치는 영향을 검토하였다. 8주 동안 단일 프로바이오틱스 B. cabrialesii sp. K1과 inulin 3%를 혼합한 신바이오틱스를 급이 한 결과 성장에서는 유의적인 차이를 보이지 않았다. 혈액학적 검사의 경우 AST, total protein 및 cholesterol에서 유의적인 차이를 보였고 ALT와 glucose는 유의적인 차이가 없었다. 비특이적 면역반응에서 는 유산균 첨가 또는 주사제로 인한 라이소자임 활성이 유익하다고 알려져 있으나 본 연구에서는 8주동안 대조구와 비교 시 유의적 차이가 없었다. 또한 B. cabrialesii K1의 경우 단일 프로바이오틱스구보다 신바이오틱스구가 대조구에 비해 NBT 활성이 증가된 것을 확인할 수 있었다. 다음 넙치의 사이토카인 발현을 확인한 결과 B. cabrialesii sp. K1를 단일(probiotics구)로 사용했을 경우 대조구와 비교 시 비장에서 유의적으로 낮게 발현되었으나 synbiotics구의 경우 비장에서 발현이 유의하게 증가되었다. 그러나 나머지 간과 장, 신장에서는 세 개의 그룹 간의 유의적 차이를 나타내지 않았다. 마지막으로 2주동안의 인위 감염을 검토한 결과 대조구는 E. tarda 95%, S. parauberis 85%, S. iniae 85%로 각각의 폐사율을 나타냈다. Probiotics구는 E. tarda 85%, S. parauberis 80%, S. iniae 80%의 폐사율을 보였고, synbiotics 구는 E. tarda 80%, S. parauberis 80%, S. iniae 85%로 각각 나타났다.


In this study, the effect of a synbiotic mixture of probiotics and 3% inulin on the growth, non-specific immune response, and mortality due to disease resistance of Paralichthys olivaceus was investigated. The results revealed that an 8-week diet of synbiotic-added feed, had no significant effect on the growth of P. olivaceus, Analysis of the hematological parameters revealed that there was a significant difference in the content of AST, total protein, and cholesterol among the groups, whereas there was no significant difference in the ALT and glucose contents. Further, there was no significant difference in the lysozyme activity of the synbiotic and control group after 8 weeks' feeding. In addition, when the symbiotic mixture was used rather than probiotics alone, the NBT activity of the synbiotic group increased compared to that of the control group. Moreover, when only probiotics were used, the cytokine expression in the spleen of the samples was significantly lower than that in the control group, whereas the expression was significantly higher in the spleen of the synbiotics group. However, there was no significant difference in the cytokine expression in the liver, intestine, and kidney of the three groups. Lastly, after injection for 2 weeks, the mortality rates of the control group towards Edwardsiella tarda, Streptococcus parauberis, and Streptococcus iniae were 95, 85 and 85%, respectively; those of the probiotics group were 85, 80, and 80%, respectively; and those of the synbiotics group were 80, 80, and 85%, respectively.

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5Anti-Inflammatory Effect of Carex scabrifolia Steud. Extract in RAW264.7 Cells

저자 : Joong Hyun Shim

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 50권 3호 발행 연도 : 2022 페이지 : pp. 354-360 (7 pages)

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This research was designed to evaluate the possible anti-inflammatory effects of Carex scabrifolia Steud. extract using RAW264.7 cells. The assessments of these effects were based on cell viability assay, mRNA expression levels of interleukin-1 alpha (IL-1α), interleukin-1 beta (IL-1β), IL-6, tumor necrosis factor alpha (TNFα), and levels of nitric oxide (NO)/prostaglandin E2 (PGE2) production. Quantitative real-time polymerase chain reaction showed that treatment with C. scabrifolia Steud. extract decreased the mRNA levels of iNOS, COX2, IL-1α, IL-1β, IL-6, and TNFα. Furthermore, from the production levels of PGE2/NO, it can be inferred that C. scabrifolia Steud. extract exhibited anti-inflammatory properties. These results suggest that C. scabrifolia Steud. extract contains anti-inflammatory compound(s), and consequently, that it may have applications as a potent cosmeceutical material.

KCI등재 SCOPUS

6Mulberry Low-Fat Ice Cream Supplemented with Synbiotic: Formulation, Phytochemical Composition, Nutritional Characteristics, and Sensory Properties

저자 : Kittisak Thampitak , Rattanaporn Pimisa , Pongsanat Pongcharoen , Suppasil Maneerat , Noraphat Hwanhlem

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 50권 3호 발행 연도 : 2022 페이지 : pp. 361-374 (14 pages)

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For this study, we designed and produced mulberry low-fat ice cream supplemented with synbiotics (MLF-ISS). The sensory characteristics and physical, chemical, and microbiological qualities of MLF-ISS were then determined. Mulberry juice inoculated with or without probiotic (Lactobacillus plantarum TISTR 926 and Saccharomyces boulardii CNCM I-745) was also tested at 37℃ for 24 h to determine probiotic growth rate, pH, total anthocyanin content (TAC), total phenolic content (TPC), and antioxidant activity (AA). Only the TAC of mulberry juice inoculated with S. boulardii CNCM I-745 increased considerably (p < 0.05) among these parameters. MLF-ISS was produced with varied mulberry fruit concentrations (0, 10, 20, 30, or 40%) (w/w). The MLF-ISS prepared with 30% mulberry fruit (w/w) (30-MLF-ISS) had a higher score in appearance, color, and sweetness (p < 0.05) when sensory qualities were measured using the 9-point hedonic scale method. In the CIE lab system (L*, a*, b*), the color values of 30-MLF-ISS were 27.80 ± 0.26, 12.99 ± 0.59, and 1.43 ± 0.05, respectively. The 30-MLF-ISS was also subjected to a proximate analysis. The melting rate of 30-MLF-ISS was 0.29 ± 0.03 g/min and the time it took for the first drop to fall was 37.00 ± 7.00 min. TAC, TPC, and AA of 30-MLF-ISS were observed to alter significantly (p < 0.05) during varied intervals of storage at -18℃ (0, 30, and 60 days). The viability of probiotics in 30-MLF-ISS slightly decreased after storage at -18℃ for 8 weeks, but remained about 6 log CFU/g. During storage at -18℃ for 0 and 120 days, no pathogenic bacteria were detected in 30-MLF-ISS. These findings show that 30-MLF-ISS has nutritional and functional value, is free of foodborne pathogenic bacteria, is safe for consumers' health, and is suitable for application in the ice cream and related food industries.

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7Evaluation of Antioxidant Potential and UV Protective Properties of Four Bacterial Pigments

저자 : Rupali Koshti , Ashish Jagtap , Domnic Noronha , Shivali Patkar , Jennifer Nazareth , Ruby Paulose , Avik Chakraborty , Pampi Chakraborty

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 50권 3호 발행 연도 : 2022 페이지 : pp. 375-386 (12 pages)

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In the present study, four distinctly colored bacterial isolates that show intense pigmentation upon brief ultraviolet (UV) light exposure are chosen. The strains are identified as Micrococcus luteus (Milky yellow), Cryseobacterium pallidum (Yellow), Cryseobacterium spp. (Golden yellow), and Kocuria turfanensis (Pink) based on their morphological and 16S rDNA analysis. Moderate salinity (1.25%), 25-37℃ temperature, and pH of 7.2 are found to be the most favorable conditions of growth and pigment production for all the selected isolates. The pigments are extracted using methanol: chloroform (1:1) and the purity of the pigments are confirmed by high-performance liquid chromatography (HPLC) and thin-layer chromatography (TLC). Further, Fourier transform infrared (FTIR) and UV-Visible spectroscopy indicate their resemblance with carotenoids and flexirubin family. The antioxidant activities of the pigments are estimated, and, all the pigments have shown significant antioxidant efficacy in 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picryl-hydrazyl (DPPH), and ferric reducing antioxidant power (FRAP) assays. The UV protective property of the pigments is determined by cling-film assay, wherein, at least 25% of UV sensitive Escherichia coli survive with bio-pigments even after 90 seconds of UV exposure compared to control. The pigments also hold a good sun protective factor (SPF) value (1.5-4.9) which is calculated with the Mansur equation. Based on these results, it can be predicted that these bacterial pigments can be further developed into a promising antioxidant and UV-protectant for several biomedical applications.

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8Anticancer and Cytotoxic Effect of Verotoxin 1 on Colon Cancer Cell Line

저자 : Mustafa Attiyah Hadid , Mohammad M. F. Al-halbosiy , Abdulwahid B. Al-shaibani

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 50권 3호 발행 연도 : 2022 페이지 : pp. 387-394 (8 pages)

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Verotoxin-1 (VT-1) or Shiga-like toxin 1 (Stx-1) is produced by enterohemorrhagic Escherichia coli (EHEC) and is an AB5 holotoxin with a strong inhibitor of protein synthesis. VT-1 is a type 2 ribosome-inactivating protein (RIP) that has been shown to have cytotoxic and anticancer potential by inducing necrosis, apoptosis, and cell cycle arrest, making it a promising antitumor candidate. Here, we tested the cytotoxicity of VT-1 on CaCo2 and NCM425 cell lines and the results showed that VT-1 was more potent on CaCo2. Morphological changes were also evaluated on the cellular level and the results showed that VT-1 caused a decrease in viable cell count, altered cell membrane permeability, and an increase in total nuclear intensity. On the other hand, VT-1 displayed a lesser impact on mitochondrial membrane potential (MMP) and cytochrome c release. On the expression of caspases 3 and 9, VT-1 exhibited an insignificant effect on both which alongside the mitochondrial membrane potential (MMP) and cytochrome c results, might indicate that CaCo2 suffered from the necrosis process as a mechanism of cell death after exposure to VT-1.

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9비피도박테리움 CBT BG7, BR3, BL3의 진세노사이드 전환능

저자 : 최지원 ( Jiwon Choi ) , 권창 ( Chang Kwon ) , 김종원 ( Jong Won Kim ) , 정명준 ( Myung Jun Chung ) , 윤종현 ( Jong Hyun Yoon ) , 임상현 ( Sanghyun Lim )

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 50권 3호 발행 연도 : 2022 페이지 : pp. 395-403 (9 pages)

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본 연구에서는 한국형 프로바이오틱스와 홍삼을 발효하여 저분자 진세노사이드인 compound K (CK)로 생물전환되는 지를 확인하였다. 프로바이오틱스 19종의 유전체 분석결과, 진세노사이드 Rb1에서 CK로 전환에 관련된 β-glucosidase는 19종 모든 균주에서 확인되었고, α-arabinofuranosidase 유전자는 3종의 균주, β-xylosidase는 6종 균주, α-rhamnosidase는 8종의 균주에서 확인되었다. 이 중 B. longum CBT BG7는 Rb1으로부터 CK까지 전환시켜, CK 함량을 증가시켰다. 또한, B. breve CBT BR3와 B. lactis CBT BL3은 Rb1을 Rd로 전환시켰다. 균체를 파쇄 또는 미파쇄하여 진세노사이드 전환 반응을 비교했을 때 미파쇄물이 F2와 CK로의 높은 전환량과 수율을 보였다. CBT BG7 + BL3와 BG7 + BR3 혼합균주는 CBT BG7 단독보다 진세노사이드 F2의 함량을 증가시켰다. CBT BG7과 α-amylase 효소를 함께 반응하였을 때에 F2 함량이 증가되었다. 본 연구는 한국형 프로바이오틱스인 CBT BG7, BR3, BL3와 홍삼을 함께 섭취할 경우, 건강에 도움을 주는 생리활성물질인 CK의 생산을 확인하였다. 추후 부탄올 등 다양한 추출용매를 활용하여 생물전환 효율 및 CK로의 전환율에 대한 추가 연구가 필요해 보인다.


In this study, we identified that the fermentation of Korean indigenous probiotics and red ginseng produced ginsenoside compound K (CK) from major ginsenosides. Based on whole genome sequencing of 19 probiotics species, β-glucosidase, α-arabinofuranosidase, β-xylosidase, and α-rhamnosidase related to bioconversion of ginsenosides are identified in the genome of 19 species, 3 species, 6 species, and 8 species, respectively. Among the 19 probiotics species, Bifidobacterium longum CBT BG7 converted from ginsenoside Rb1 to CK, and both B. breve CBT BR3 and B. lactis CBT BL3 converted ginsenoside Rb1 to Rd. The final concentration and yield of ginsenoside F2 and CK were higher in the fermentation with the nondisrupted cells than with disrupted cells. The combination of both CBT BG7 and BL3, and CBT BG7 and BR3 showed higher amounts of F2 than CBT BG7 only. CBT BG7 with adding α-amylase increased the amounts of F2. In this study, we identified that the fermentation of both Korean indigenous probiotic bacteria CBT BG7, BR3 and BL3, and red gingseng is able to produce CK, a bioactive compound that promotes health benefits.

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10A Culture-Independent Comparison of Microbial Communities of Two Maturating Craft Beers Styles

저자 : Joao Costa , Isabel N. Sierra-garcia , Angela Cunha

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 50권 3호 발행 연도 : 2022 페이지 : pp. 404-413 (10 pages)

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The process of manufacturing craft beer involves a wide variety of spontaneous microorganisms, acting in different stages of the brewing process, that contribute to the distinctive characteristics of each style. The objective of this work was to compare the structure of microbial communities associated with two different craft beer styles (Doppelbock and Märzen lagers), at a late maturation stage, and to identify discriminative, or style-specific taxa. Bacterial and fungal microbial communities were analyzed by Illumina sequencing of 16S rRNA gene of prokaryotes and the ITS 2 spacer of fungi (eukaryotes). Fungal communities in maturating beer were dominated by the yeast Dekkera, and by lactic acid (Lactobacillus and Pediococcus) and acetic acid (Acetobacter) bacteria. The Doppelbock barrels presented more rich and diverse fungal communities. The Märzen barrels were more variable in terms of structure and composition of fungal and bacterial communities, with occurrence of exclusive taxa of fungi (Aspergillus sp.) and bacteria (L. kimchicus). Minority bacterial taxa, differently represented in the microbiome of each barrel, may underlie the variability between barrels and ultimately, the distinctive traits of each style. The composition of the microbial communities indicates that in addition to differences related to upstream stages of the brewing process, the contact with the wood barrels may contribute to the definition of style-specific microbiological traits.

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1Isolation of Novel Strains of Lactobacillus gasseri EJL and Bifidobacterium breve JTL from Breast Milk and Infant Feces: A Longitudinal Study of a Mother-infant Pair

저자 : Heetae Lee , Chong-kil Lee , Kyungjae Kim

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 1호 발행 연도 : 2021 페이지 : pp. 1-8 (8 pages)

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Human breast milk is a potential source of bacteria for the development of the intestinal microbiota of infants. Several species within the genera Lactobacillus and Bifidobacterium were demonstrated to shape the gut microbiota of infants. In this study, the bacterial diversity was investigated in the breast milk and feces of a mother-infant pair, and probiotic candidates were identified. Importantly, the novel L. gasseri EJL and B. breve JTL strains were isolated from breast milk and infant feces samples, respectively; their completed genome was resolved using de novo sequencing. In addition, the bacterial composition in the infant's feces at 1 week revealed the prevalence of Bifidobacterium and Streptococcus; a higher diversity was observed after 3 weeks. In particular, the abundance of Akkermansia was sharply increased at 7 weeks, further increasing thereafter, up to 15 weeks. Our results suggest that human breast milk and infant's feces are a source of probiotic candidates.

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2Characterization of the Recombinant Glutamate Decarboxylase of Lactobacillus brevis G144 Isolated from Galchi Jeotgal, a Korean Salted and Fermented Seafood

저자 : Jeong A Kim , Ji Yeong Park , Jeong Hwan Kim

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 1호 발행 연도 : 2021 페이지 : pp. 9-17 (9 pages)

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A γ-aminobutyric acid (GABA)-producing microorganism was isolated from galchi (hairtail fish, Trichiurus lepturus) jeotgal, a Korean salted and fermented seafood. The G144 isolate produced GABA excessively when incubated in MRS broth containing monosodium glutamate (MSG, 3%, w/v). G144 was identified as Lactobacillus brevis through 16S rRNA and recA gene sequencing. gadB and gadC encoding glutamate decarboxylase (GAD) and glutamate/GABA antiporter, respectively, were cloned and gadB was located downstream of gadC. The operon structure of gadCB was confirmed by reverse transcription (RT)-polymerase chain reaction. gadB was overexpressed in Escherichia coli and recombinant GAD was purified and its size was 54.4 kDa as evidenced by SDS-PAGE results. Maximum GAD activity was observed at pH 5.0 and 40℃ and the activity was dependent on pyridoxal 5'-phophate. The Km and Vmax of GAD were 8.6 mM and 0.01 mM/min, respectively.

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3Lactobacillus plantarum G72 Showing Production of Folate and Short-chain Fatty Acids

저자 : Hye Ji Jang , Na-kyoung Lee , Hyun-dong Paik

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 1호 발행 연도 : 2021 페이지 : pp. 18-23 (6 pages)

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The aim of this study was to determine the production of folate, short chain fatty acids (SCFAs), and antimicrobial activity exhibited by Lactobacillus plantarum G72 for potential dietary application in pregnant women. L. plantarum G72 has been reported to possess characteristic activities and functionality including β-galactosidase activity and antioxidant activities. L. plantarum G72 showed antibacterial activity against pathogenic bacteria (Listeria monocytogenes ATCC 15313, Salmonella typhimurium P99, Escherichia coli ATCC 25922, and Staphylococcus aureus KCCM 11335) using a modified method, and formation of the largest inhibition zone was observed against S. aureus KCCM 11335 (12.0-17.0 mm). The adherence of four food-borne pathogenic bacteria to HT-29 cells was inhibited by L. plantarum G72 (0.13 to 0.92 log CFU/ml). The most considerable inhibition of adherence to HT-29 cells was observed by using L. plantarum G72 against S. typhimurim P99. Additionally, folate production by L. plantarum G72 was 50.1 ng/ml, and L. plantarum G72 produced relatively more lactic acid (11,176.73 mg/kg) than acetic, propionic, or butyric acids. Therefore, the results of this study suggest that L. plantarum G72 may serve as a multifunctional food additive in the health industry.

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4Melanogenesis Inhibitory Activity of Epicatechin-3-O-Gallate Isolated from Polygonum amphibium L.

저자 : Young Kyung Lee , Buyng Su Hwang , Yong Hwang , Seung Young Lee , Young Taek Oh , Chul Hwan Kim , Hyeon Ju Nam , Yong Tae Jeong

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 1호 발행 연도 : 2021 페이지 : pp. 24-31 (8 pages)

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This study aimed to investigate the melanogenesis inhibitory activity of epicatechin-3-O-gallate (ECG) isolated from Polygonum amphibium L. ECG was isolated from the ethanol extract of P. amphibium L, and its chemical structure was determined using spectroscopic methods such as LC-ESI-MS, 1D-NMR, and UV spectroscopy. ECG inhibited the melanogenesis of B16F10 cells in a dose-dependent manner. Particularly, it decreased the melanin content by 27.4% at 200 μM concentration, compared with the control, in B16F10 cells, without causing cytotoxicity. It is noteworthy that the expression of three key proteins, including tyrosinase, tyrosinase-related protein-1 (TRP-1), TRP-2, and microphthalmia-associated transcription factor (MITF), involved in melanogenesis, is significantly inhibited by ECG. The ECG isolated in this study caused the inhibition of body pigmentation and tyrosinase activity in vivo in the zebrafish model. These results suggest that the ECG isolated from P. amphibium L. is an effective anti-melanogenesis agent.

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5Screening of Myxobacteria Carrying Tubulysin Biosynthetic Genes

저자 : Hyesook Hyun , Juo Choi , Daun Kang , Yungpil Kim , Pilgoo Lee , Gregory J. Y. Chung , Kyungyun Cho

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 1호 발행 연도 : 2021 페이지 : pp. 32-38 (7 pages)

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Tubulysins are a group of secondary metabolites produced by myxobacteria that inhibit the function of the eukayotic cytoskeleton. We developed a pair of PCR primers that specifically amplified tubulysin biosynthetic genes. Using these primers, eight out of the eighty-one strains of myxobacteria belonging to the Cystobacteraceae family that harbored putative tubulysin biosynthetic genes were screened through PCR analysis. The selected strains included two Archangium gephyra, two Stigmatella sp., two Vitiosangium cumulatum, and two unidentified myxobacteria. LC-MS analysis of the culture extracts from the selected strains revealed that A. gephyra KYC4066 produced putative tubulysin A and B.

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6The in vitro and in vivo Safety Evaluation of Lactobacillus acidophilus IDCC 3302

저자 : Won Yeong Bang , Seung A Chae , O-hyun Ban , Sangki Oh , Chanmi Park , Minjee Lee , Minhye Shin , Jungwoo Yang , Young Hoon Jung

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 1호 발행 연도 : 2021 페이지 : pp. 39-44 (6 pages)

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As consumption of healthy foods continues to garner remarkable public attention, ensuring probiotic safety has become a priority. In this study, the safety of Lactobacillus acidophilus IDCC 3302 was assessed in vitro and in vivo. L. acidophilus IDCC 3302 showed negative results for hemolytic and β-glucuronidase activities. The whole-genome analysis (WGA) revealed that L. acidophilus IDCC 3302 did not possess antibiotic resistance or virulence genes. The minimal inhibitory concentrations of L. acidophilus IDCC 3302 confirmed its safety concerning antibiotic resistance. Furthermore, L. acidophilus IDCC 3302 was demonstrated to be nontoxic in the oral toxicity test in rats. Therefore, the results suggested that L. acidophilus IDCC 3302 might be safe for human consumption.

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7Studies on Improved Amylases Developed by Protoplast Fusion of Aspergillus species

저자 : Tolulope Modupe Adeleye , Sharafadeen Olateju Kareem , Bankole Mobolaji Olufunmilayo , Olusegun Atanda , Michael Bamitale Osho , Olawale Dairo

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 1호 발행 연도 : 2021 페이지 : pp. 45-56 (12 pages)

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Improved amylases were developed from protoplast fusants of two amylase-producing Aspergillus species. Twenty regenerated fusants were screened for amylase production using Remazol Brilliant Blue agar. Crude enzyme extracts produced by solid state fermentation of rice bran were assayed for activity. Three variable factors (temperature, pH and enzyme type) were optimized to increase the amylase activity of the parents and selected fusants using rice bran medium and solid state fermentation. Analysis of this optimization was completed using the Central Composite Design (CCD) of the Response Surface Methodology (RSM). Amylase activity assays conducted at room temperature and 80℃ demonstrated that Aspergillus designates, T5 (920.21 U/ml, 966.67 U/ml), T13 (430 U/ml, 1011.11 U/ml) and T14 (500.63 U/ml, 1012.00 U/ml) all exhibited improved function making them the preferred fusants. Amylases produced from these fusants were observed to be active over the entire pH range evaluated in this study. Fusants T5 and T14 demonstrated optimal activity under acidic and alkaline conditions, respectively. Fusants T13 and T14 produced the most amylase at 72 h while parents TA, TC and fusant T5 produced the most amylase after 96 h of incubation. Response surface methodology examinations revealed that the enzyme from fusant T5 was the optimal enzyme demonstrating the highest activity (1055.17 U/ml) at pH 4 and a temperature of 40℃. This enzyme lost activity with further increases in temperature. Starch hydrolysis using fusant T5 gave the highest yield of glucose (1.6158 g/100 ml). The significant activities of the selected fusants at 28 ± 2℃ and 80℃ and the higher sugar yields from cassava starch hydrolysis over their parental strains indicate that it is possible to improve amylase activity using the protoplast fusion technique.

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8헴철이 풍부한 영양원이 혐기성 세균의 생장과 생존에 미치는 영향: 락토바실러스 가세리 모델연구

저자 : 이승기 ( Seungki Lee ) , 김필 ( Pil Kim )

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 1호 발행 연도 : 2021 페이지 : pp. 57-64 (8 pages)

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Lactic acid bacteria (LAB), belonging to the Firmicutes phylum, lack heme biosynthesis and, thus, are characterized as fermentative and catalase-negative organisms. To verify the hypothesis that heme-rich-nutrients might compensate the heme-biosynthesis incapability of non-respiratory LAB in animal gut, a heme-rich-nutrient was fed to a dog and its fecal microbiome was analyzed. Firmicutes abundance in the feces from the heme-rich-nutrient-fed dog was 99%, compared to 92% in the control dog. To clarify the reason of increased Firmicutes abundance in the feces from the heme-rich-nutrient-fed dog, Lacobacillus gasseri were used as model anerobic LAB to study a purified heme (hemin). The anaerobic growth of L. gasseri in the medium with 25 μM hemin supplementation was faster than that in the medium without hemin, while the growth in the 50 μM hemin-supplemented medium did not vary. Cellular activities of the cytochrome bd complex were 1.55 ± 0.19, 2.11 ± 0.14, and 2.20 ± 0.08 U/gcell in the cells from 0, 25, and 50 μM hemin-supplemented medium, while intracellular ATP concentrations were 7.90 ± 1.12, 11.95 ± 0.68, and 12.56 ± 0.58 μmolATP/gcell, respectively. The ROS-scavenging activities of the L. gasseri cytosol from 25 μM and 50 μM hemin-supplemented medium were 68% and 82% greater than those of the cytosol from no hemin supplemented-medium, respectively. These findings indicate that external hemin could compensate the heme-biosynthesis incapability of L. gasseri by increasing the cytosolic ROS-scavenging and extra ATP generation, possibly through increasing the electron transfer. Increase in the number of anaerobic bacteria in heme-rich-nutrient-fed animal gut is discussed based on the results.

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9Exploring the Catalytic Significant Residues of Serine Protease Using Substrate-Enriched Residues and a Peptidase Inhibitor

저자 : Zahoor Khan , Maryam Shafique , Amir Zeb , Nusrat Jabeen , Sehar Afshan Naz , Arif Zubair

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 1호 발행 연도 : 2021 페이지 : pp. 65-74 (10 pages)

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Serine proteases are the most versatile proteolytic enzymes with tremendous applications in various industrial processes. This study was designed to investigate the biochemical properties, critical residues, and the catalytic potential of alkaline serine protease using in-silico approaches. The primary sequence was analyzed using ProtParam, SignalP, and Phyre2 tools to investigate biochemical properties, signal peptide, and secondary structure, respectively. The three-dimensional structure of the enzyme was modeled using the MODELLER program present in Discovery Studio followed by Molecular Dynamics simulation using GROMACS 5.0.7 package with CHARMM36m force field. The proteolytic potential was measured by performing docking with casein- and keratin-enriched residues, while the effect of the inhibitor was studied using phenylmethylsulfonyl fluoride, (PMSF) applying GOLDv5.2.2. Molecular weight, instability index, aliphatic index, and isoelectric point for serine protease were 39.53 kDa, 27.79, 82.20 and 8.91, respectively. The best model was selected based on the lowest MOLPDF score (1382.82) and DOPE score (-29984.07). The analysis using ProSA-web revealed a Z-score of -9.7, whereas 88.86% of the residues occupied the most favored region in the Ramachandran plot. Ser327, Asp138, Asn261, and Thr326 were found as critical residues involved in ligand binding and execution of biocatalysis. Our findings suggest that bioengineering of these critical residues may enhance the catalytic potential of this enzyme.

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10Heterologous Expression of Interferon α-2b in Lactococcus lactis and its Biological Activity against Colorectal Cancer Cells

저자 : Lita Meilina , Sri Budiarti , Apon Zaenal Mustopa , Huda Shalahudin Darusman , Lita Triratna , Muhammad Ajietuta Nugraha , Muhammad Sabiq Bilhaq , Ratih Asmana Ningrum

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 1호 발행 연도 : 2021 페이지 : pp. 75-87 (13 pages)

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Type I Interferons (IFNα) are known for their role as biological anticancer agents owing to their cell-apoptosis inducing properties. Development of an appropriate, cost-effective host expression system is crucial for meeting the increasing demand for proteins. Therefore, this study aims to develop codon-optimized IFNα-2b in L. lactis NZ3900. These cells express extracellular protein using the NICE system and Usp45 signal peptide. To validate the mature form of the expressed protein, the recombinant IFNα-2b was screened in a human colorectal cancer cell line using the cytotoxicity assay. The IFNα-2b was successfully cloned into the pNZ8148 vector, thereby generating recombinant L. lactis pNZ8148-SPUsp45-IFNα-2b. The computational analysis of codon-optimized IFNα-2b revealed no mutation and amino acid changes; additionally, the codon-optimized IFNα-2b showed 100% similarity with native human IFNα-2b, in the BLAST analysis. The partial size exclusion chromatography (SEC) of extracellular protein yielded a 19 kDa protein, which was further confirmed by its positive binding to anti-IFNα-2b in the western blot analysis. The crude protein and SEC-purified partial fraction showed IC50 values of 33.22 μg/ml and 127.2 μg/ml, respectively, which indicated better activity than the metabolites of L. lactis NZ3900 (231.8 μg/ml). These values were also comparable with those of the regular anticancer drug tamoxifen (105.5 μg/ml). These results demonstrated L. lactis as a promising host system that functions by utilizing the pNZ8148 NICE system. Meanwhile, codon-optimized usage of the inserted gene increased the optimal protein expression levels, which could be beneficial for its large-scale production. Taken together, the recombinant L. lactis IFNα-2b is a potential alternative treatment for colorectal cancer. Furthermore, its activity was analyzed in the WiDr cell line, to assess its colorectal anticancer activities in vivo.

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