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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
  • : 1219


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1Acyl Homoserine Lactone in Interspecies Bacterial Signaling

저자 : Poonam Kanojiya , Rajashri Banerji , Sunil D. Saroj

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 50권 1호 발행 연도 : 2022 페이지 : pp. 1-14 (14 pages)

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Bacteria communicate with each other through an intricate communication mechanism known as quorum sensing (QS). QS regulates different behavioral aspects in bacteria, such as biofilm formation, sporulation, virulence gene expression, antibiotic production, and bioluminescence. Several different chemical signals and signal detection systems play vital roles in promoting highly efficient intra- and interspecies communication. Gram-negative bacteria coordinate gene regulation through the production of acyl homoserine lactones (AHLs). Gram-positive bacteria do not code for AHL production, while some gram-negative bacteria have an incomplete AHL-QS system. Despite this fact, these microbes can detect AHLs owing to the presence of LuxR solo receptors. Various studies have reported the role of AHLs in interspecies signaling. Moreover, as bacteria live in a polymicrobial community, the production of extracellular compounds to compete for resources is imperative. Thus, AHL-mediated signaling and inhibition are considered to affect virulence in bacteria. In the current review, we focus on the synthesis and regulation mechanisms of AHLs and highlight their role in interspecies bacterial signaling. Exploring interspecies bacterial signaling will further help us understand host-pathogen interactions, thereby contributing to the development of therapeutic strategies intended to target chronic polymicrobial infections.

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2Anti-Inflammatory Effects of the Chemical Compounds Obtained from Celastrus hindsii in RAW264.7 Cells

저자 : Thi Ngoc Anh Pham , Hae Lim Kim , Suwon Oh , Seung Hwan Yang

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 50권 1호 발행 연도 : 2022 페이지 : pp. 15-21 (7 pages)

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Celastrus hindsii (family Celastraceae) is located abundantly in the United States, China, and Vietnam, where it is utilized as a traditional herbal and traditional drug for the care of cancer. However, the antioxidant and anti-inflammatory effects of Celastrus hindsii extract are unknown. In our research, the antioxidant activity of Celastrus hindsii leaf extract was investigated, and then anti-inflammatory efficacy of C. hindsii extract in lipopolysaccharide (LPS)-induced RAW264.7 cells. First, our results revealed that C. hindsii extract showed powerful antioxidant capability. Moreover, the application of C. hindsii extract significantly reduced nitric oxide (NO) production without cytotoxic effects. Furthermore, C. hindsii extract reduced the generation of pro-inflammatory cytokines, like as interleukin (IL)-6, IL-1β, and TNF-α. Our results are the first to confirm the anti-inflammatory capability of C. hindsii extract in RAW264.7 cells.

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3Anti-Inflammatory Effects of Abalone (Haliotis discus hannai) Viscera via Inhibition of ROS Production in LPS-Stimulated RAW 264.7 Cells

저자 : Tai-sun Shin , Kap Seong Choi , Jiyeon Chun , Kang-hee Kho , Seon Ah Son , Sun-yup Shim

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 50권 1호 발행 연도 : 2022 페이지 : pp. 22-30 (9 pages)

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Haliotis discus hannai called abalone, is the valuable marine mollusks and the by-products of abalone processing are viscera. Brownish abalone male viscera (AMV), which have not been reported as having anti-inflammatory effects, was extracted with acetone and fractionated by different six acetone/hexane ratios (0, 10, 20, 30, 40, and 100%) using a silica column via in vitro ABTS and DPPH radical and nitric oxide (NO) production assay-guided fractionation. Among the fractions, the acetone/hexane ratio 40%, A40 exhibited the most potent radical scavenging activities and inhibition of lipopolysaccharide (LPS)-induced NO production without cytotoxicity. A40 inhibited LPS-induced intracellular reactive oxygen species (ROS) production in a dose-dependent manner. Western blot analysis revealed that A40 down-regulated the activation of NF-κB, MAPK (ERK 1/2, p-38, and JNK), and inflammatory enzymes, inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2. Moreover, this fraction inhibited the generation of pro-inflammatory cytokines such as interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α. These results suggested that AMV containing A40 with anti-inflammatory and anti-oxidantive effects, is the effective therapeutic and functional material for treating inflammatory disorders.

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4Melanin 생성 인자 억제 효과를 통한 Mangifera indica L. Peel의 미백효과 연구

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

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 50권 1호 발행 연도 : 2022 페이지 : pp. 31-39 (9 pages)

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본 연구의 목적은 애플망고 껍질의 열수 및 70% 에탄올 추출물의 미백 효과를 검증하고자 하였다. 애플망고 껍질의 열수 및 에탄올 추출물의 미백 효과를 측정하기 위해 tyrosinase 저해 활성을 측정한 결과, 최종 농도인 1,000 μg/ml에서 열수 추출물은 9%, 에탄올 추출물은 35%의 저해 효과를 보였다. 세포를 통해 미백효과를 측정하고자 멜라노마 세포인 B16-F10을 이용해 세포 생존율을 MTT assay를 사용하여 측정하였다. 세포 생존율 측정 결과, 100 μg/ml 농도에서 각각 95.64%, 103.36%의 세포 생존율을 나타내었다. 이후 실험은 세포 생존율이 95% 이상 나타난 농도인100 μg/ml 이하의 농도에서 실험을 수행하였다. 미백 효과는 멜라닌 합성에 관여하는 인자의 단백질 및 mRNA 발현을 측정하여 결정하였다. 단백질 발현은 western blot을 이용하여 측정하였으며, 그 결과 MITF, tyrosinase, TRP-1 및 TRP-2에 대한 단백질 발현은 100 μg/ml 농도에서 열수 추출물에 의해 59%, 65%, 26%, 18% 감소하였고, 에탄올 추출물에 의해 64%, 40%, 18%, 52% 감소하였다. MITF, Tyrosinase, TRP-1 및 TRP-2의 mRNA 발현은 RT-PCR을 통해 확인하였으며, 그 결과 100 μg/ml에서 열수 추출물에 의해 27%, 44%, 40%, 22% 감소하였고, 에탄올 추출물에 의해 9%, 51%, 11%, 52% 감소하였다. 따라서 애플망고 껍질 추출물이 미백 효과가 있음을 확인하였고, 천연물 소재로서의 이용 가치가 높을 것으로 사료되어진다.


The purpose of this study was to investigate the whitening effects of hot water (AMPW) and ethanol (AMPE) extracts of Mangifera indica L. peel. To verify the whitening effects, tyrosinase inhibitory activity was measured. 9.51% inhibitory activity, and 35.98% inhibitory activity at 1,000 μg/ml. The effects of AMPW and AMPE on cell viability were measured using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay in B16-F10 melanoma cells. Greater than 95% cell viability was observed at 100 μg/ml. Thus, subsequent experiments were performed at concentrations less than 100 μg/ml. The whitening effects were confirmed by measuring the protein and mRNA expression levels of microphthalmia-associated transcription factor, tyrosinase, tyrosinase-related protein 1 (TRP-1), and TRP-2, which are factors involved in melanin synthesis. Western blotting and reverse transcription-polymerase chain reaction results confirmed that 100 μg/ml AMPW and AMPE showed superior inhibitory effects than the control treatment (alpha-melanocyte stimulating hormone only). Therefore, Mangifera indica L. peel extract had a whitening effect, and thus, has potential as a natural material for use in cosmetics.

KCI등재 SCOPUS

5Mycorrhizal Synthesis of Perigord Black Truffle (Tuber melanosporum) with Mexican Oak Species

저자 : Gonzalo Guevara-guerrero , Giovanni Pacioni , Marco Leonardi , Fortunato Garza Ocañas , Rigoberto Gaitán Hernández

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 50권 1호 발행 연도 : 2022 페이지 : pp. 40-50 (11 pages)

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The Périgord black truffle (Tuber melanosporum) is an edible fungus and among the most expensive foods worldwide. It is the basis of a multimillion-dollar bio-business. Truffle farming does not exist in Mexico, and no formal studies have been conducted on its culture. This report describes the mycorrhizal synthesis (i.e., artificial union of fungus with host) of T. melanosporum with oak species native to Mexico (Quercus polymorpha, Q. fusiformis, and Q. canbyi). The mycorrhizal association was successful in Q. polymorpha and Q. fusiformis, as confirmed morphologically and using T. melanosporum molecular primers (ITSML/ ITS4LNG). The effect of the ectomycorrhizal fungus on host growth (stem diameter) was statistically significant. Illustrations of the study are presented.

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6Potential Probiotic Characteristics and Safety Assessment of Lactobacillus rhamnosus SKG34 Isolated from Sumbawa Mare's Milk

저자 : I Nengah Sujaya , Gede Ngurah Rsi Suwardana , Kazuyoshi Gotoh , I Wayan Sumardika , Komang Ayu Nocianitri , Ni Putu Sriwidyani , I Wayan Gede Artawan Eka Putra , Masakiyo Sakaguchi , Ni Nengah Dwi Fatmawati

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 50권 1호 발행 연도 : 2022 페이지 : pp. 51-62 (12 pages)

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Lactobacillus rhamnosus SKG34 (LrSKG34), a potential probiotic strain, was successfully isolated from Sumbawa Mare's milk. Our previous studies showed that the strain is resistant to gastrointestinal conditions, possesses antioxidant activity, and lowers blood cholesterol levels. Further clarification of the potential probiotic characteristics and safety assessment are necessary. This study aimed to evaluate the adhesion of LrSKG34 to Caco-2 cell monolayers and its effect on mucosal integrity in vitro. We also examined the LrSKG34 safety profile based on antimicrobial susceptibility testing, haemolytic activity determination, Caco-2 cell monolayer translocation evaluation, and in vivo investigation of the effect of LrSKG34 on the physiology, biochemical markers, and histopathological appearance of major organs in an animal model. LrSKG34 attached to Caco-2 cell monolayers and maintained mucosal integrity in vitro. The typical resistance of lactobacilli to ciprofloxacin, gentamicin, vancomycin, trimethoprim-sulfamethoxazole, and metronidazole was confirmed for LrSKG34. No haemolytic activity was observed on blood agar plates, and no LrSKG34 translocation was observed in Caco-2 cell monolayers. Administration of LrSKG34 to Sprague-Dawley rats did not adversely affect body weight. No abnormalities in hematological parameters, serum biochemistry levels, or histopathological structures of major organs were observed in LrSKG34-treated rats. Collectively, the results implicate LrSKG34 as a promising and potentially safe probiotic candidate for further development.

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7Characterization of Two Cryptic Plasmids from Levilactobacillus zymae GU240

저자 : Huong Giang Le , Min Jae Kim , Hye Sung Jeon , Ji Yeon Yoo , Yun Ji Kang , Tae Jin Kim , Jeong Hwan Kim

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 50권 1호 발행 연도 : 2022 페이지 : pp. 63-70 (8 pages)

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Two small cryptic plasmids, pHG1 and pHG2, were isolated from Levilactobacillus zymae (formerly Lactobacillus zymae) GU240 and characterized. pHG1 is 1,814 bp in size with a GC content of 37.4% and contains two open reading frames. orf1 can potentially encode a protein of 101 amino acids (aa) with 99% identity with the copy number control protein of Lacticaseibacillus paracasei. orf2 can potentially encode a protein of 230 aa with 99% identity with a replication protein from multiple species. Six inverted repeats (IR I-VI) and six direct repeats (DR I-VI) were found in pHG1. pHG2 is 2,864 bp in size, with a GC content of 39.6%. pHG2 has two orfs. orf1 might encode a protein with 99% identity with the TrsL transmembrane protein. orf2 might encode a protein with 99% identity with plasmid recombination proteins from lactic acid bacteria. Both pHG1 and pHG2 may be useful as frames for constructing lactic acid bacteria-Escherichia coli shuttle vectors.

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8Purification, Characterization and Application of a Cold Active Lipase from Marine Bacillus cereus HSS

저자 : Sahar Wm. Hassan , Hala H. Abd El Latif , Ehab A. Beltagy

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 50권 1호 발행 연도 : 2022 페이지 : pp. 71-80 (10 pages)

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Lipases (triacylglycerol acylhydrolases [EC 3.1.1.3]) are water-soluble enzymes. They catalyze the hydrolysis of fats and oils. A cold-active lipase from marine Bacillus cereus HSS, isolated from the Mediterranean Sea, Alexandria, Egypt, was purified and characterized. The total purification depending on lipase activity was 438.9 fold purification recording 632 U/mg protein. The molecular weight of the purified lipase was estimated to be 65 kDa using sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. The optimum substrate concentration, enzyme concentration, pH, and temperature were 1.5 mM, 100 μl, pH 6 and 10℃, respectively. The lipase was tolerant to NaCl concentrations ranging from 1.5 to 4.5%. The lipase was affected by the tested metal ions, and its activity was inhibited by 16% in the presence of 0.05 M SDS. The application of the cold-active lipase for the removal of an oil stain from a white cotton cloth showed that it is a promising biological agent for the treatment of oily wastes and other related applications. To the best of our knowledge, this is the first report of the purification and characterization of a lipase from marine B. cereus HSS isolated from the Mediterranean Sea.

KCI등재 SCOPUS

9Molecular Identification and Technological Properties of Acetic Acid Bacteria Isolated from Malatya Apricot and Home-Made Fruit Vinegars

저자 : Eda Buyukduman , Hatice Ebrar Kirtil , Banu Metin

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 50권 1호 발행 연도 : 2022 페이지 : pp. 81-88 (8 pages)

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Acetic acid bacteria (AAB) are versatile organisms involved in the production of variety of fermented foods, such as vinegar and kombucha, and products of biotechnological relevance, such as bacterial cellulose. In the present study, Malatya apricot, a variety with protected designation of origin (PDO), and vinegar samples produced using various fruits were used to isolate AAB. The 19 AAB isolates obtained were typed using (GTG)5 fingerprinting, and the ones selected were identified by sequencing either 16S rDNA alone or in combination with 16S-23S rRNA internal transcribed spacer region or ligA gene. While all apricot isolates (n = 10) were Gluconobacter cerinus, vinegar isolates (n = 9) were composed of Komagataeibacter saccharivorans, Acetobacter syzygii, and possible two new species of AAB, Komagataeibacter sp., and Gluconobacter sp. (GTG)5 fingerprinting showed the presence of several genotypes of G. cerinus in the apricot samples. Screening for some technologically relevant properties, including thermotolerance, ethanol tolerance, and cellulose production capability, showed that all Komagataeibacter and some Gluconobacter isolates could tolerate the temperature of 35℃, and that vinegar isolates could tolerate up to 8% ethanol. One isolate, Komagataeibacter sp. GUS3 produced bacterial cellulose (1 g/l) and has the potential to be used for cellulose production.

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10Triclosan Resistant Bacteria from Sewage Water: Culture Based Diversity Assessments and Co-Resistance Profiling to Other Antibiotics

저자 : Muhmmad Salman , Noor Ul Bashar , Uzma Kiran , Zuhra Shafiq , Fareesa Khan , Raees Khan , Farrukh Hussain , Sudhair Abbas Bangash , Yasin Ahmad , Shabir Ahmad

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 50권 1호 발행 연도 : 2022 페이지 : pp. 89-94 (6 pages)

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Triclosan (TCS) is an antimicrobial agent used in various human personal care products against both gram-positive and gram-negative bacteria. The purpose of this study was to evaluate the presence of TCS-resistant bacteria in sewage water in Peshawar, Khyber Pakhtunkhwa (KPK), Pakistan, for the first time. TCS-supplemented Luria Bertani (LB) agar was used to isolate TCS-tolerant bacteria. A total of 17 TCS-resistant isolates were randomly selected from a large pool of bacteria that showed growth on TCS-supplemented LB agar. Based on gram staining and physiochemical characteristics, the isolated strains were identified as Salmonella typhi (n = 6), Escherichia coli (n = 4), Citrobacter freundii (n = 4), Proteus mirabilis (n = 1), Enterobacter cloacae (n = 1), and Pseudomonas aeruginosa (n = 1). The Triclosan mean minimum inhibitory concentrations (MICs) for the isolates of Salmonella typhi, Escherichia coli, Citrobacter freundii, Proteus mirabilis, Enterobacter cloacae, and Pseudomonas aeruginosa were 23.66 μg ml-1, 18.75 μg ml-1, 42 μg ml-1, 32 μg ml-1, 64 μg ml-1, and 128 μg ml-1, respectively. The antibiogram revealed that all isolates were resistant to penicillin G (100%) and linezolid (100%), followed by ampicillin (94%), tetracycline (76%), tazobactam (76%), sulbactam/cefoperazone (64%), polymyxin PB (58%), amikacin (29.41%), aztreonam (29.41%), imipenem (5%), and gentamicin (5%). This is the first known study regarding the isolation of TCS-tolerant bacteria from sewage water in Peshawar, KPK, Pakistan. It was concluded that all the TCS-resistant isolates were multidrug resistant (MDR) gram-negative rod-shaped bacteria, mostly belonging to the Enterobacteriaceae family.

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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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