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한국미생물·생명공학회지 update

Microbiology and Biotechnology Letters

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  • : 자연과학분야  >  생물
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  • : 산업미생물학회지(~2001)→한국미생물·생명공학회지(2002~)

수록정보
수록범위 : 30권1호(2002)~49권3호(2021) |수록논문 수 : 1,185
한국미생물·생명공학회지
49권3호(2021년 09월) 수록논문
최근 권호 논문
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KCI등재 SCOPUS

1나고야의정서 이행에 따른 새로운 유전자원 접근 이익공유 체계의 이해와 미생물 연구자의 대응방안

저자 : 이종현 ( Jonghyun Lee ) , 안민호 ( Minho An ) , 장영효 ( Young-hyo Chang )

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 3호 발행 연도 : 2021 페이지 : pp. 269-282 (14 pages)

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최근 생명공학 R&D의 필수 재료인 유전자원에 대한 취득과 그 이용을 규제하는 국제 규범, 즉 나고야의정서가 등장하면서 관련 분야 연구자에게 상당한 불편과 부담이 우려되고 있다. 나고야의정서가 발효함에 따라 그 동안 인류공동유산으로서 마치 공공재처럼 모든 국가가 자유롭게 사용해왔던 유전자원에 대하여 자원 보유국의 배타적 소유권이 인정되면서 자원의 취득과 이용에 대한 각국의 법적 규제가 도입되고 있다. 특히 유전자원의 해외의존도가 높은 우리나라는 보다 체계적인 대응을 해야 할 필요성이 있다. 이 글은 연구자를 포함한 국내외 유전자원 이용자의 나고야의정서에 대한 인식 제고를 위하여, 먼저 의정서의 주요 핵심내용을 분석하고 해외 유전자원 이용을 위한 접근, 취득 및 이익공유의 전체적인 구조와 흐름을 제시함으로써 향후 적절한 대응에 도움이 될 수 있도록 구성·기술하였다.
나고야의정서에 적절하게 대처하기 위해서는 연구자 본인의 노력과 국가적인 차원에서의 지원이 동시에 필요하다. 우선 연구자는 나고야의정서 체제에서의 연구활동 진행에 관한 전체적인 틀과 각 단계별 구체적인 대응에 관한 이해를 반드시 해야 한다. 이를 토대로 유전자원 접근단계부터 이로 인해 발생한 이익의 공유단계까지 제공국의 ABS절차에 적절하게 대응해야 한다. 이렇게 볼 때 나고야의정서로 인해 과거 대비 연구활동에 일정부분 제한이 가해지고 연구 외적인 부담이 가중되었다고 할 수 있다. 하지만 전 세계적으로 유전자원에 대한 국가의 주권을 인정하는 시류가 한시적이라고 보기 힘든 상황임은 분명하다. 나고야의정서 체제를 중심으로 자국의 유전자원에 대한 주권적 권리를 지키고자 하는 자원부국의 기조가 더 강해질 것으로 예상된다. 따라서 국내 연구자도 관련 피해를 최소화하기 위한 대비를 보다 확실히 해야 할 것이다.


Following enforcement of the Nagoya Protocol (NP), in which the sovereign rights to genetic resources of countries are recognized, new legal obligations regarding access and benefit-sharing (ABS) that did not exist before have now been imposed on researchers. To implement the NP, many countries are introducing new procedures and regulations when a researcher wants to obtain genetic resources for commercial or noncommercial uses. It is therefore expected that resource-rich countries will adopt strong regulations to protect their genetic resources. In this regard, Korean microbiologists will need to respond to these changes to minimize the potential damages caused by the ABS. This paper reviews the key contents of the NP to raise its awareness among scientific researchers and further presents specific measures to meet the ABS obligations accordingly. For example, Korean researchers, in principle, do not need to acquire Prior Informed Consents (PICs) when they access Korean microbial resources for both commercial or research purposes. Nevertheless, when a foreign culture collection agency such as DSMZ requests a confirmation of compliance with the NP to deposit genetic resources, Korean researchers can also apply for a PIC with the Korean government as an exception. By referring to this article, microbiologists will be able to conduct their research in compliance with the NP while respecting the legal ABS obligations of each resource-providing country.

KCI등재 SCOPUS

2굴피나무 잎 추출물의 위암세포에 대한 세포사멸 유도 효과

저자 : 이형선 ( Hyeong-seon Lee )

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 3호 발행 연도 : 2021 페이지 : pp. 283-288 (6 pages)

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본 연구에서는 굴피나무 잎 추출물로부터 사람 위암세포 주인 AGS 세포에 대한 apoptosis 가 유도되어 항암 활성을 가지는지를 확인하고자 한다. AGS 세포에 굴피나무 잎 추출물을 농도별 (0, 50, 150, 200 pg/ml)로 처리하여 세포에서 나타나는 양상들을 확인하였다. MTS 측정으로 암세포 생존율을 확인한 결과 굴피나무 잎 추출물에 따라 농도의존적으로 세포에 독성을 보였으며, 이러한 세포 죽음이 apoptosis 유도에 의한 것인지를 annexin V/FITC-PI 염색을 통해 확인하였다. 그 결과, early apoptosis를 보이는 세포의 양상이 유의성 있게 증가하였다. 굴피나무 잎 추출물의 처리는 세포 주기에서 sub-Gl 기의 증가로 암세포가 더 이상 분열하지 못하고 증식이 억제됨을 보여준다. 내인성 경로에 관여하는 Bax, Bak, Bcl-2, Bcl-xL의 증감을 mRNA 수준에서 RT- PCR로 확인한 결과, Bax, Bak가 증가하고, Bcl-2, Bcl-xL이 감소를 동반하며 미토콘드리아를 통한 세포사멸의 신호전달 경로가 진행되고 있음을 확인할 수 있었다. 이러한 결과를 종합하면 굴피나무 잎 추출물은 사람 위암세포에 대한 항암 활성을 가지는 효능적 가치가 있음에 가능성을 제시할 수 있을 것으로 생각되며,추가적인 동물실험을 통한 인체적용시험의 효능 검증을 통해 의약품 개발 가능성을 확인해야 한다.


This study investigated the anticancer activity of methanol extract from Platycarya strobilacea leaf in AGS human gastric cancer cells. We determined the cell viability effect of P. strobilacea using MTS assay. Apoptosis induction and cell cycle arrest were confirmed by fluorescein isothiocyanate and propidium iodide staining using cellometer K2. The mRNA expression levels of the Bcl-2 family were confirmed by reverse transcription-polymerase chain reaction. The cell viability was decreased in a dose-dependent manner treated with different concentrations of P. strobilacea. Total, early, and late apoptotic cells were dramatically increased, and the cell cycle was arrested at the sub-G1 phase. The mRNA expressions of Bcl-2 and Bcl-xL were reduced, whereas pro-apoptotic factors, Bax and Bak, were increased in a dose-dependent manner. These results suggested that P. strobilacea leaf extract induced significant apoptotic activity through an intrinsic mitochondria pathway.

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3Cloning and Expression of a Fibrinolytic Enzyme Gene, aprECJ1, from Bacillus velezensis CJ1 Isolated from Myeolchi Jeotgal

저자 : Ji Yeon Yoo , Zhuang Yao , Se Jin Lee , Hye Sung Jeon , Jeong Hwan Kim

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 3호 발행 연도 : 2021 페이지 : pp. 289-297 (9 pages)

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Bacillus velezensis CJ1, showing significant fibrinolytic activity, was isolated from Myeolchi Jeotgal, a popular Korean fermented seafood. When B. velezensis CJ1 was grown on four different culture media, the culture on the Luria-Bertani (LB) broth showed the highest fibrinolytic activity (102.94 mU/μl) at 48 h. LB was also the best medium for growth. SDS-PAGE of culture supernatant showed four major bands, 38, 35, 27, and 22 kDa in size. Fibrin zymography showed four active bands, 50, 47, 40, and 30 kDa in size. A gene homologous to aprE of the Bacillus species was cloned by PCR. DNA sequencing showed that aprECJ1 can encode a protease consisting of 382 amino acids. The translated amino acid sequence of AprECJ1 showed high identity values with those of B. velezensis strains and other Bacillus species. The aprECJ1 gene was introduced into B. subtilis WB600 using an E. coli-Bacillus shuttle vector, pHY300PLK, and overexpressed. A 27 kDa band corresponding to the mature form of AprECJ1 was produced and confirmed by SDS-PAGE and fibrin zymography. B. subtilis WB600 [pHYaprECJ1] showed 1.8-fold higher fibrinolytic activity than B. velezensis CJ1 at 48 h.

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4무-유도인자 단백질 발현 시스템을 이용한 재조합 시아노박테리아의 광합성 스쿠알렌 생산 평가

저자 : 최선영 ( Sun Young Choi ) , 우한민 ( Han Min Woo )

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 3호 발행 연도 : 2021 페이지 : pp. 298-304 (7 pages)

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Photosynthetic conversion through cyanobacteria and microalgae is an increasingly serious concern in the global warming crisis. Many value-added substances are produced through strain improvement, and much research and development is being conducted to determine its potential as an actual industrial strain. Economic barriers throughout processing production can be overcome to produce value-added chemicals by microalgal strains. In this study, we engineered cyanobacteria strains for the photosynthetic production of squalene and confirmed the continuous cultivation of CO2 and light conditions. The free-inducer system of gene expression was developed at the cyanobacterial strains. Then, the squalene production level and growth of the recombinant cyanobacteria were analyzed and discussed. For bio solar-cell factories, the ability to regulate genes based on the free-inducer gene expression system promotes metabolic engineering research and construction to produce value-added chemicals.

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5Molecular Cloning, Protein Expression, and Regulatory Mechanisms of the Chitinase Gene from Spodoptera littoralis Nucleopolyhedrovirus

저자 : Norhan Yasser , Reda Salem , Maha Alkhazindar , Ismail A. Abdelhamid , Said A. S. Ghozlan , Wael Elmenofy

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 3호 발행 연도 : 2021 페이지 : pp. 305-315 (11 pages)

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The cotton leafworm, Spodoptera littoralis, is a major pest in Egypt and many countries worldwide, and causes heavy economic losses. As a result, management measures to control the spread of the worm are required. S. littoralis nucleopolyhedrovirus (SpliNPV) is one of the most promising bioagents for the efficient control of insect pests. In this study, a chitinase gene (chitA) of a 1.8 kb DNA fragment was cloned and fully characterized from SpliNPV-EG1, an Egyptian isolate. A sequence of 601 amino acids was deduced when the gene was completely sequenced with a predicted molecular mass of 67 kDa for the preprotein. Transcriptional analyses using reverse transcription polymerase chain reaction (RT-PCR) revealed that chitA transcripts were detected first at 12 h post infection (hpi) and remained detectable until 168 hpi, suggesting their transcriptional regulation from a putative late promoter motif. In addition, quantitative analysis using quantitative RT-PCR showed a steady increase of 7.86-fold at 12 hpi in chitA transcription levels, which increased up to 71.4-fold at 120 hpi. An approximately 50 kDa protein fragment with chitinolytic activity was purified from ChitA-induced bacterial culture and detected by western blotting with an antirecombinant SpliNPV chitinase antibody. Moreover, purification of the expressed ChitA recombinant protein showed in vitro growth inhibition of two different fungi species, Fusarium solani and F. oxysporum, confirming that the enzyme assembly and activity was correct. The results supported the potential role and application of the SpliNPV-ChitA protein as a synergistic agent in agricultural fungal and pest control programs.

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6Detection of AluI Endonuclease Activity by Using Double Stranded DNA-Templated Copper Nanoclusters

저자 : Ji Su Yang , Jongback Gang

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 3호 발행 연도 : 2021 페이지 : pp. 316-319 (4 pages)

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Restriction endonucleases play an important role in molecular cloning, clinical diagnosis, and pharmacological drug studies. In this study, DNA-templated copper nanoclusters (DNA-CuNCs) were used to detect AluI endonuclease activity due to their high fluorescence emission and rapid synthesis of DNA-CuNCs under ambient conditions. Results showed that AluI activity was detected in a highly sensitive manner at low concentrations of AluI endonuclease by the fluorescence intensity of DNA-CuNCs. Additionally, its inhibition was monitored in the presence of daidzein under optimal conditions.

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7Preparation of High GABA-Enriched Yeast Extract by Non-Saccharomyces Yeasts Isolated from Korean Traditional Fermented Soybean Product

저자 : Nho-eul Song , Da-bin Lee , Seon-hye Lee , Sang-ho Baik

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 3호 발행 연도 : 2021 페이지 : pp. 320-328 (9 pages)

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High GABA-enriched yeast extract, for various nutritionally and pharmaceutically important functional foods, was prepared using a novel isolate of Debaryomyces hansenii JBCC541. Under optimized conditions, GABA conversion rates are significantly enhanced up to 7.55 g/l by D. hansenii JBCC541, increasing their synthesis yield 40 times. The total amino acid content of the prepared yeast extract was 10733.86 mg/l (257.36 mg/g), consisting of alanine, lysine, glutamine, leucine, and valine as the primary amino acids. The GABA content was significantly enhanced up to 6790 mg/l (162.80 mg/g) in the presence of glutamic acid, with approximately 10-fold higher GABA production. Flavor amino acids were also highly enhanced, indicating that the prepared yeast extract might be useful for preparing various functional and sensuous foods. Our results were promising as a GABA-enriched yeast extract preparation tool ensuring a suitable food material level with the potential for functionally enhanced food industrial applications.

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8해양미생물 Cellulophga sp. J9-3이 생산하는 베타-아가레이즈의 분리 및 생화학적 특성

저자 : 김다솜 ( Da Som Kim ) , 김종희 ( Jong-hee Kim ) , 지원재 ( Won-jae Chi )

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 3호 발행 연도 : 2021 페이지 : pp. 329-336 (8 pages)

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Cellulophga sp. J9-3은 셀룰로스 분해능력을 갖으며, Flavobacteriaceae 과에 속하는 그람-음성 호기성 해양 세균이다. 또한, J9-3 균주는 고체 및 액체 배지에서 한천을 가수 분해 할 수 있으며, 배지에 첨가한 아가로스(agarose)에 의해 아가레이즈(agarase)의 생산이 현저하게 유도되는 특성을 보였다. Cellulophga sp. J9-3의 세포 배양액으로부터, 황산암모늄 침전 및 3 단계의 컬럼 크로마토그래피를 연속적으로 수행하여, 한 개의 agarase 단백질, AgaJ93을 순수하게 정제하였다. 정제된 AgaJ93은 아가로스에 대한 분해 활성이 가장 강하였으며, starch에 대해서도 아가로스 대비 약 22% 정도의 분해 활성을 나타냈다. AgaJ93은 아가로스를 분해하여 사이즈가 큰 올리고당 중간체를 경유하여, 최종산물로 네오아가로테트라오스와 네오아가로헥사오스까지 분해함을 확인하였으며, 이는 AgaJ93이 endo-type β-아가레이즈 임을 의미한다. AgaJ93은 pH 7.0, 35℃에서 최대 활성을 나타냈고, Co2+ 이온에 의해 6배 이상의 활성증가를 보였다. AgaJ93의 N-terminal sequence 분석 결과, AgaJ93은 Cellulophaga sp. W5C의 내열성 endo-type β-아가레이즈 Aga2와 82%의 상동성을 보였으나, 두 효소의 생화학적 특성이 달랐다. 따라서, AgaJ93은 기존에 보고된 β-아가레이즈 들과는 다른 신규의 아가레이즈 일 것으로 예상된다.


Cellulophga sp. J9-3, is a gram-negative, aerobic marine bacterium belonging to the family Flavobacteriaceae. In addition to cellulose degradability, the J9-3 strain is also capable of hydrolyzing agar in the solid and liquid medium, and the production of agarase in the presence of agarose can be remarkably induced by the bacterium. From the cell culture broth of Cellulophga sp. J9-3, ammonium sulfate precipitation and three kinds of column chromatography were successively performed to purify a specific agarase protein, the AgaJ93. Purified AgaJ93 showed the strongest hydrolyzing activity towards agarose (approximately 22%), and even displayed activity towards starch. AgaJ93 hydrolyzed agarose into neoagarotetraose and neoagarohexaose via various oligosaccharide intermediates, indicating that AgaJ93 is an endo-type β-agarase. AgaJ93 showed maximum activity at a pH of 7.0 and temperature of 35 °C. Its activity increased by more than six times in the presence of Co2+ ions. The N-terminal sequence of AgaJ93 showed 82% homology with the heat-resistant endo-type β-agarase Aga2 of Cellulophaga sp. W5C. However, the biochemical properties of the two enzymes were different. Therefore, AgaJ93 is expected to be a novel agarose, different from the previously reported β-agarases.

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9Optimization of Medium for Lipase Production from Zygosaccharomyces mellis SG1.2 Using Statistical Experiment Design

저자 : Marisa Dian Pramitasari , Miftahul Ilmi

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 3호 발행 연도 : 2021 페이지 : pp. 337-345 (9 pages)

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Lipase (triacylglycerol lipase, EC 3.1.1.3) is an enzyme capable of hydrolyzing triacylglycerol, to produce fatty acids and glycerol and reverse the reaction of triacylglycerol synthesis from fatty acids and glycerol through transesterification. Applications of lipase are quite widespread in the industrial sector, including in the detergent, paper, dairy, and food industries, as well as for biodiesel synthesis. Lipases by yeasts have attracted industrial attention because of their fast production times and high stability. In a previous study, a lipase-producing yeast isolate was identified as Zygosaccharomyces mellis SG1.2 and had a productivity of 24.56 U/mg of biomass. This productivity value has the potential to be a new source of lipase, besides Yarrowia lypolitica which has been known as a lipase producer with a productivity of 0.758 U/mg. Lipase production by Z. mellis SG1.2 needs to be increased by optimizing the production medium. The aims of this study were to determine the significant component of the medium for lipase production and methods to increase lipase production using the optimum medium. The two methods used for the statistical optimization of production medium were Taguchi and RSM (Response Surface Methodology). The data obtained were analyzed using Minitab 18 and SPSS 23 software. The most significant factors which affected lipase productivity were olive oil and peptones. The optimum medium composition consisted of 1.02% olive oil, 2.19% peptone, 0.05% MgSO4·7H2O, 0.05% KCl, and 0.2% K2HPO4. The optimum medium was able to increase the lipase productivity of Z. mellis SG1.2 to 1.8-fold times the productivity before optimization.

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10Effect of Lactic Fermentation and Spray Drying Process on Bioactive Compounds from Ngoc Linh Ginseng Callus and Lactobacillus plantarum Viability

저자 : Lieu My Dong , Nguyen Thi Thuy Linh , Nguyen Thi Hoa , Dang Thi Kim Thuy , Do Dang Giap

발행기관 : 한국미생물생명공학회 간행물 : 한국미생물·생명공학회지 49권 3호 발행 연도 : 2021 페이지 : pp. 346-355 (10 pages)

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Ngoc Linh ginseng is one of the most valuable endemic medicinal herbs in Vietnam. In this study, Ngoc Linh ginseng callus was fermented by Lactobacillus plantarum ATCC 8014 (at 6, 7, and 8 log CFU/ml) to evaluate the extraction efficiency of bioactive compounds. The post-fermentation solution was spray-dried using maltodextrin with or without Stevia rebaudiana (3% and 6% v/v) as the wall material. Bioactive compounds such as polyphenols, polysaccharides, and total saponins, and L. plantarum viability during fermentation and after spray-drying, as well as under simulated gastric digestion, were evaluated in this study. The results showed that probiotic density had a significant effect on bioactive compounds, and L. plantarum at 8 log CFU/ml showed the best results with a short fermentation time compared to other tests. The total content of polyphenols, polysaccharides, and saponins reached 5.16 ± 0.18 mg GAE/g sample, 277.2 ± 6.12 mg Glu/g sample, and 4.17 ± 0.15 mg/g sample, respectively after 20 h of fermentation at the initial density of L. plantarum (8 log CFU/ml). Although there was no difference in the particle structure of the preparation, the microencapsulation efficiency of the bioactive compound in the samples containing S. rebaudiana was higher than that with only maltodextrin. The study also indicated that adding S. rebaudiana improved the viability of L. plantarum in gastric digestion. These results showed that S. rebaudiana, a component stimulating probiotic growth, combined with maltodextrin as a co-prebiotic, improved the survival rate of L. plantarum in simulated gastric digestion.

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