• 한국미생물·생명공학회지
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• 제38권4호
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• pp.475-480
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• 2010

Actinomycetes are Gram-positive soil bacteria and one of the most important industrial microorganisms due to superior biosynthetic capabilities of many valuable secondary metabolites as well as production of various valuable bioconversion enzymes. Among them are cytochrome P450 hydroxylase (CYP), which are hemoproteins encoded by a super family of genes, are universally distributed in most of the organisms from all biological kingdoms. Actinomycetes are a rich source of soluble CYP enzymes, which play critical roles in the bioactivation and detoxification of a wide variety of metabolite biosynthesis and xenobiotic transformation. Cyclosporin A (CyA), one of the most commonly-prescribed immunosuppressive drugs, was previously reported to be hydroxylated at the position of 4th N-methyl leucine by a rare actinomycetes called Sebekia benihana, leading to display different biological activity spectrum such as loss of immunosuppressive activities yet retaining hair growth-stimulating side effect. In order to improve this regio-selective CyA hydroxylation in S. benihana, previously-identified several secondary metabolite up-regulatory genes from Streptomyces coelicolor and S. avermitilis were heterologously overexpressed in S. benihana using an $ermE^*$ promoter-containing Streptomyces integrative expression vector. Among tested, SCO4967 encoding a conserved hypothetical protein significantly stimulated region-specific CyA hydroxylation in S. benihana, implying that some common regulatory systems functioning in both biosynthesis and bioconversion of secondary metabolite might be present in different actinomycetes species.

• Fisheries and Aquatic Sciences
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• 제17권2호
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• pp.215-222
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• 2014

To explore marine microorganisms with medical potential, we isolated and identified marine bacteria from floats, marine algae, animals, and sponges collected from Jeju Island, Korea. We isolated and identified 21 different strains from the marine samples by 16S rRNA analysis, cultured them in marine broth, and extracted them with ethyl acetate (EtOAc) to collect secondary metabolite fractions. Next, we evaluated their anti-oxidative and anti-inflammatory effects. Among the 21 strains, the secondary metabolite fraction of Bacillus badius had both strong antioxidant and anti-inflammatory activity, and thus was selected for further experiments. An antioxidant compound detected from the secondary metabolite fraction of B. badius was purified by preparative centrifugal partition chromatography (n-hexane:EtOAc:methanol:water, 4:6:4:6, v/v), and identified as diolmycin A2. Additionally, diolmycin A2 strongly inhibited nitric oxide production. Thus, we successfully identified a significant bioactive compound from B. badius among the bacterial strains collected from Jeju Island.

• Plant Biotechnology Reports
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• 제4권2호
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• pp.109-116
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• 2010

Morinda citrifolia adventitious roots were cultured in shake flasks using Murashige and Skoog medium with different types and concentrations of auxin and cytokinin. Root (fresh weight and dry weight) accumulation was enhanced at 5 $mg\;l^{-1}$ indole butyric acid (IBA) and at 7 and 9 $mg\;l^{-1}$ naphthalene acetic acid (NAA). On the other hand, 9 $mg\;l^{-1}$ NAA decreased the anthraquinone, phenolic and flavonoid contents more severely than 9 $mg\;l^{-1}$ IBA. When adventitious roots were treated with kinetin (0.1, 0.3 and 0.5 $mg\;l^{-1}$) and thidiazuron (TDZ; 0.1, 0.3 and 0.5 $mg\;l^{-1}$) in combination with 5 $mg\;l^{-1}$ IBA, fresh weight and dry weight decreased but secondary metabolite content increased. The secondary metabolite content (including 1,1-diphenyl-2-picrylhydrazyl activity) increased more in TDZ-treated than in kinetin-treated roots. Antioxidative enzymes such as catalase (CAT) and guaiacol peroxidase (G-POD), which play important roles in plant defense, also increased. A strong decrease in ascorbate peroxidase activity resulted in a high accumulation of hydrogen peroxide. This indicates that adventitious roots can grow under stress conditions with induced CAT and G-POD activities and higher accumulations of secondary metabolites. These results suggest that 5 $mg\;l^{-1}$ IBA supplementation is useful for growth and secondary metabolite production in adventitious roots of M. citrifolia.

• Journal of Microbiology and Biotechnology
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• 제23권3호
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• pp.297-303
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• 2013

Phylogenetic analysis of the groEL1 and xynB1 gene sequences from Sorangium cellulosum strains isolated in Korea previously revealed the existence of at least 5 subgroups (A-E). In the present study, we used sequence analysis of polymerase chain reaction-amplified biosynthetic genes of strains from the 5 subgroups to indicate correlations between S. cellulosum subgroups and their secondary metabolic gene categories. We detected putative biosynthetic genes for disorazol, epothilone, ambruticin, and soraphen in group A, group C, group D, and group E strains, respectively. With the exception of KYC3204, culture extracts from group A, group B, and group C strains exhibited no noticeable antimicrobial inhibitory activities. By contrast, culture extracts from group D strains inhibited the growth of Candida albicans, whereas culture extracts from group E strains inhibited the growth of C. albicans and Staphylococcus aureus. High performance liquid chromatography analysis of the culture extracts from the strains of each subgroup revealed unique peak patterns. Our findings indicate the existence of at least 5 subgroups of S. cellulosum strains, each of which has the potential to produce a unique set of secondary metabolites.

• 생물환경조절학회지
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• 제32권3호
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• pp.217-225
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• 2023

본 실험은 마이크로웨이브 처리가 상추 유묘의 생육 변화와 이차대사산물 함량의 변화를 알아보고자 수행되었다. 파종 후 3주째 상추 유묘에 2.45GHz 주파수와 200W의 마이크로웨이브를 0, 4, 8 및 12초 동안 처리하고, 4주간 식물공장에서 재배한 후 생육 및 성분 분석을 수행하였다. 지하부와 지상부의 생체중과 건물중, 엽면적, 엽장 및 엽수는 마이크로웨이브 처리시간이 증가할수록 감소하였다. 4초 처리구와 비교하여 12초 처리구에서 chlorophyll a, chlorophyll b 및 총 carotenoids의 함량이 증가되었으며 총 페놀 함량은 감소하였다. 무처리구와 비교하여 8초 처리구에서 총 플라보노이드 함량이 감소하였다. 이러한 결과들은 산화적 스트레스에 의해 이차대사산물 함량이 변화된 것으로 사료된다. 총 플라보노이드 함량을 제외한 이차대사산물 함량은 각 처리구에서 무처리구와 비교하여 유의한 차이가 없었지만, 각 처리구 사이의 유의한 차이는 200W와 2.45GHz의 마이크로웨이브 처리가 4주 후 상추의 이차대사산물 함량에 영향을 줄 수 있다는 것을 시사한다.

• 한국결정학회지
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• 제16권2호
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• pp.66-74
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• 2005

다양한 생리활성을 가지는 marine natural products는 일반적인 secondary metabolites와 유사한 구조를 가지는데, 염소, 브롬, 요오드의 할로겐 원소에 의해 수식이 되어있는 것이 일반적이다. Vanadium haloperoxidase는 이러한 marine natural products의 생산에 중요한 효소로 vanadate를 조효소로 하는 금속효소이다. 본 리뷰에서는 vanadium haloperoxidase의 분리와 단백질 구조를 살펴보고, 이 금속효소의 작용기작에 대해서 설명할 것이다. 마지막으로, vanadium haloperoxidase의 반응성과 secondary metabolites 중 indole, terpenoids, acetogenins의 생합성 예를 살펴볼 것이다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVII)
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• pp.373-374
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• 2005

• 한국미생물·생명공학회지
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• 제34권3호
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• pp.211-215
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• 2006

S. lividans에서 클로닝된 조절유전자인 afsR2를 과발현시 나타나는 up-regulated protein과 down-regulated protein 관련 유전자들을 proteomics 방법으로 선별하였고[3], 이를 방선균 발현벡터인 pSE34를 이용하여 제작된 pPNP, pGPD를 S. avermitilis에 형질전환시켜 avermectin 및 이차대사산물의 생산량 차이를 비교 분석하였다. 그 결과 up-regulated protein으로 예상되던 PNP는 wild-type S. avermitilis에서만 제한적으로 avermetin-type 대사산물의 생산성 향상을 촉진시켰으며, 이와 반대로 down-regulated protein으로 예상되었던 GPD는 avermectin-type 대사산물의 생산량을 wild type S. avermitilis에서는 4배, over-producer S. avermitilis에서는 2.5배 증가시켰다. 본 연구결과는, 방선균 조절 단백질들이 이차대사산물의 종류 및 생합성 기작에 따라 전혀 다르게 작용될 수 있음을 암시하며, 향후 이들 조절 단백질들에 대한 보다 구체적인 분자수준에서의 연구 필요성을 제시하고 있다.

• Journal of Microbiology and Biotechnology
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• 제17권12호
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• pp.2066-2070
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• 2007

It was found that Shinorhizobium meliloti hemoprotein (SM) was more effective than Vitreoscilla hemoglobin (Vhb) in promoting secondary metabolites production when overexpressed in Streptomyces lividans TK24. The transformant with sm (sm-transformant) produced 2.7-times and 3-times larger amounts of actinorhodin than the vhb-transformant in solid culture and flask culture, respectively. In both solid and flask cultures, a larger amount of undecylprodigiocin was produced by the sm-transformant. It is considered that the overexpression of SM especially has activated the pentose phosphate pathway through oxidative stress, as evidenced by an increased NADPH production observed, and that it has promoted secondary metabolites biosynthesis.

• Journal of Microbiology and Biotechnology
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• 제25권8호
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• pp.1265-1274
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• 2015

Secondary metabolite-based chemotaxonomic classification of Streptomyces (8 species, 14 strains) was performed using ultraperformance liquid chromatography-quadrupole-time-offlight-mass spectrometry with multivariate statistical analysis. Most strains were generally well separated by grouping under each species. In particular, S. rimosus was discriminated from the remaining sevens pecies (S. coelicolor, S. griseus, S. indigoferus, S. peucetius, S. rubrolavendulae, S. scabiei, and S. virginiae) in partial least squares discriminant analysis, and oxytetracycline and rimocidin were identified as S. rimosus-specific metabolites. S. rimosus also showed high antibacterial activity against Xanthomonas oryzae pv. oryzae, the pathogen responsible for rice bacterial blight. This study demonstrated that metabolite-based chemotaxonomic classification is an effective tool for distinguishing Streptomyces spp. and for determining their species-specific metabolites.