CoQ10 생성 세균의 선별 및 발효조건 최적화

정근일,강원화,이정아,신동하,배경숙,박호용,박희문,Jeong, Keun-Il,Kang, Won-Hwa,Lee, Jung-Ah,Shin, Dong-Ha,Bae, Kyung-Sook,Park, Ho-Young,Park, Hee-Moon 한국미생물학회 2010 미생물학회지 Vol.46 No.1

Coenzyme Q10 (CoQ10)은 전자전달계에 필수적인 요소로 질병치료 및 완화에 도움이 되어 산업 의학적으로 그 활용도가 넓어지고 있다. 본 연구에서는 새로운 CoQ10 생산균주를 선별하기 위하여 quinone 분석 결과 CoQ10을 함유하는 것으로 확인된 8종 미생물의 생장특성과 CoQ10 생산능을 1차 조사하여, 세균류인 Paracoccus denitrificans KCTC 2530과 Asaia siamensis KCTC 12914를 대량배양을 통한 CoQ10 생산에 유리한 특성을 갖는 균주로 선별하였다. 이들 세균류의 생장 및 CoQ10 생산의 최적조건을 플라스크배양으로 조사한 결과, M81 배지를 기반으로 하여 탄소원으로는 4% fructose, 질소원으로는 2% yeast extract가 가장 좋은 것으로 조사되었으며, 배양온도는 $30^{\circ}C$, 배지의 최적 pH는 P. denitrificans KCTC 2530의 경우 pH 6.0, A. siamensis KCTC 12914의 경우 pH 8.0으로 조사되었다. 이를 바탕으로 2 L fed-batch culture를 수행한 결과, P. denitrificans KCTC 2530은 1 L 당 $14.34{\pm}0.473$ mg, A. siamensis KCTC 12914는 $12.53{\pm}0.231$ mg의 CoQ10을 생산하였다. Coenzyme Q10 (CoQ10) is an essential lipid-soluble component of membrane-bound electron transport chains. CoQ10 is involved in several aspects of cellular metabolism and is increasingly being used in therapeutic applications for several diseases. Despite the recent accomplishments in metabolic engineering of Escherichia coli for CoQ10 production, the production levels are not yet competitive with those by fermentation or isolation. So we tested several microorganisms obtained from the KCTC of Biological Resource Center to find novel sources of strain-development for CoQ10-production. Then we selected two strains, Paracoccus denitrificans (KCTC 2530) and Asaia siamensis (KCTC 12914), and tested to optimize the CoQ10 production conditions. Among the carbon sources tested, CoQ10 production was the highest when fructose was supplied about 4% concentration. Yeast extract produced the highest CoQ10 production about 2% concentration. The highest CoQ10 production was obtained at pH 6.0 for P. denitrificans and pH 8.0 for A. siamensis. And two strains showed the highest CoQ10 production at $30^{\circ}C$, but the highest DCW was obtained at $37^{\circ}C$. In the fed-batch culture, P. denitrificans yielded $14.34{\pm}0.473$ mg and A. siamensis yielded $12.53{\pm}0.231$ mg of final CoQ10 production.

토양에서 분리한 Bacillus Siamensis BC-S09의 항진균 활성

서민경 ( Min-kyoung Seo ),양현주 ( Hyun-joo Yang ),남효송 ( Hyo-song Nam ) 한국환경농학회 2021 한국환경농학회 학술대회집 Vol.2021 No.-

A bacterium having antifungal activity was isolated from soil in Gokseong-gun, South Korea and identified as Bacillus siamensis. Antifungal bacteria were isolated from various soil samples. The samples were suspended in phosphate-buffered saline (PBS), spread on Tryptic Soy agar (TSA) medium and allowed to grow at 30℃ for 24 h under aerobic condition. The strain BC-S09 exhibiting the highest antifungal activity was identified as Bacillus siamensis. The strain BC-S09 was identified by 16S rRNA gene sequence analysis after polymerase chain reaction (PCR) amplification using the universal primers 27F and 1492R as described Stackebrandt and Goodfellow. The 16S RNA sequences were compared using “16S-based ID” in the EzBioCloud (https://www.ezbiocloud.net) database. The phylogenetic analysis was performed using the neighbor-joining method in the MEGA program (https://www. megasoftware.net/). Various fungi were kept on potato-dextrose agar (PDA) slants at 25℃. Spore inoculums were prepared by growing the molds on 10-days-old PDA slants and collected after vigorously shaking slants with sterile peptone water (0.2% w/v). Bacterial isolates exhibiting antifungal activity were observed directly on PDA agar plates where the indicator molds test microorganisms were placed with control and experimental group. Bacteria and fungi was incubation of 25℃ for 2 day to 7 day in aerobic incubator. The results showed that Bacillus siamensis BC-S09 suppressed the growth of phytopathogenic fungi including Colletotrichum coccodes (KACC40010), Colletotrichum acutatum (KACC40689), Colletotrichum acutatum (KACC40804 - strong virulence), Altermaria citri (KACC42128), Fusarium oxysporum, Rhizoctonia solani.

Radula Tooth Structure of Eight Bithyniid Snails Observed by Scanning Electron Microscopy

Kim, Jae Jin 한국패류학회 2015 The Korean Journal of Malacology Vol.31 No.1

우리나라를 비롯한 아시아, 유럽, 호주 등에서 채집한 쇠우렁이 과 패류 8종 (Bithynia striatula, B. tentaculata, B. siamensis, B. leachi, B. kiusiuensis, B. misella, Bithynia sp., Gabbia australis) 을 대상으로 주사전자현미경 (SEM) 을 이용하여 치설을 관찰한 바 다음과 같은 결과를 얻었다. 치설열의 수는 B. siamensis와 B. leachi의 경우 60-70개로 다른 종들의 치설열의 수 40-50개보다 많았다. 관찰한 8종의 패류의 치설 구조는 좌우대칭으로 한 치설열에 7개의 치아가 있었다. 즉 1개의 중치를 중심으로 좌우에 각각 1개의 측치와 2개의 외연치가 분포하였다. 따라서 기본 치설식은 2:1:1:1:2로 표시할 수 있었다. 소첨두의 수와 모양은 개체별, 종별로 차이가 있었으며 중치에만 기저돌기가 분포하였다. 중치의 중앙돌기의 구조는 B. striatula, B. tentaculata, B. siamensis 및 Bithynia sp.의 구조가 다른 종들과 차이가 있었다. Eight species of the family Bithyniidae, Bithynia striatula, B. tentaculata, B. siamensis, B. leachi, B. kiusiuensis, B. misella, Bithynia sp. from Nepal, and Gabbia australis were studied for comparison of the radula structure by scanning electron microscope. Radular ribbons of B. siamensis and B. leachi had 60-70 rows, and the other species had 40-50 rows. Teeth of all species studied had symmetrical structure, i.e. a total of seven teeth (one central tooth, and one lateral and two pairs of marginal teeth on each side) in each row were observed. The basic tooth formula of eight bithyniids was 2:1:1:1:2. Numbers and shapes of cusps were slightly differ from individuals or species. The mesocone of the central tooth of B. striatula, B. tentaculata, B. siamensis and Bithynia sp. differed from those of the other species observed.

Structural variations generated by simian foamy virus-like (SFV) in Crocodylus siamensis

Panupon Twilprawat,김송미,Kornsorn Srikulnath,Kyudong Han 한국유전학회 2017 Genes & Genomics Vol.39 No.10

Endogenous retrovirus (ERV) integrates into the germline of its host and could remain in the genome as a molecular fossil. ERV is one of sources that cause INDEL and recombination events in the vertebrate genomes, leading to various genomic and genetic changes in their hosts. There have been many studies conducted on ERVs in the vertebrate genomes to elucidate their evolutionary history. However, ERVs have not been studied well in Crocodylus siamensis. Here, we report structural variations among SFV1 elements (simian foamy virus-like), ERVs in C. siamensis. We initially identified 26 SFV1 candidates in the genome and experimentally verified 9 SFV1_1 and 5 SFV1_10 elements using PCR display. Their structural analyses showed that most of them are solitary-LTRs but two SFV1_1 elements are full-length. Through further analyses, we found that the two full-length elements retain intact ORFs. We examined transcription factor binding sites within their LTR sequences to predict promoter/enhancer activities. In sum, we identified 14 crocodile-specific SFV1 elements and the results of their structural analyses suggest that they could contribute to genomic or phenotypic variations in C. siamensis population.

Complete mitochondrial genome of a kind of snakehead fish Channa siamensis and its phylogenetic consideration

Rui Li,Gang Wang,Zheng‑Yong Wen,Yuan‑Chao Zou,Chuan‑Jie Qin,Yu Luo,Jun Wang,Gui‑Hong Chen 한국유전학회 2019 Genes & Genomics Vol.41 No.2

The snakehead fish, Channa siamensis, belongs to the genus of Channa (perciformes: Channidae) and was first reported by Günther in 1861. Despite it has been described approximately for 15 decades, the genetic information is limited and the taxon status of this kind of fish is still unclear. The primary objective of this study is to get more genomic data and calculate the taxon location of this kind of fish. The next generation sequencing method was used to obtain the whole mitochondrial DNA information, and bioinformatic analysis was performed to investigate the evolutionary status and taxon location of C. siamensis. The circular mitochondrial DNA was 16,570 bp in length, and which showed typical piscine structure and arrangement. The overall nucleotide composition was 29.28% A, 24.72% T, 30.71% C, 15.29% G, with 54.1% AT, respectively. Phylogenetic analyses using concatenated amino acid and nucleotide sequences of the 13 protein-coding genes with two different methods (Maximum likelihood and Bayesian analysis) both highly supported C. siamensis belongs to the genus Channa and shows a close relationship with C. micropeltes. These data will provide more useful information for a better understanding of the mitochondrial genomic diversities and evolution in fish as well as novel genetic markers for studying population genetics and species identification.

Screening of Antagonistic Bacillus against Brown Rot in Dendrocalamus latiflorus and Preparation of Applying Bacterial Suspension

Fengying Luo,Hang Chen,Wenjian Wei,Han Liu,Youzhong Chen,Shujiang Li 한국식물병리학회 2024 Plant Pathology Journal Vol.40 No.1

The aim of this study was to isolate biocontrol bacteria that could antagonize brown rot of <i>Dendrocalamus latiflorus</i>, optimize the culture conditions, and develop an effective biocontrol preparation for brown rot of <i>D. latiflorus</i>. This study isolated a bacterium with an antagonistic effect on bamboo brown rot from healthy <i>D. latiflorus</i> rhizosphere soil. Morphology, molecular biology, and physiological biochemistry methods identified it as <i>Bacillus siamensis</i>. The following culturing media and conditions improved the inhibition effect of <i>B. siamensis</i>: the best culturing media were 2% sucrose, 1.5% yeast extract, and 0.7% potassium chloride; the optimal culturing time, temperature, pH, and inoculation amount were 48 h, 30℃, 6, and 20%. The optimum formula of the applying bacterial suspension was 14% sodium dodecyl benzene sulfonate emulsifier, 4% Na₂HPO₄·2H₂O, 0.3% hydroxypropyl methylcellulose thickener, and 20% <i>B. siamensis</i>. The pot experiment results showed the control effect of applying bacterial suspension, diluted 1,000 times is still better than that of 24% fenbuconazole suspension. The applying bacterial suspension enables reliable control of brown rot in <i>D. latiflorus</i>.

Potentiality of Beneficial Microbe Bacillus siamensis GP-P8 for the Suppression of Anthracnose Pathogens and Pepper Plant Growth Promotion

Woo Ji Min,Kim Hyun Seung,In Kyu Lee,Byeon Eun Jeong,Chang Won Jun,이윤수 한국식물병리학회 2024 Plant Pathology Journal Vol.40 No.4

This study was carried out to screen the antifungal activity against Colletotrichum acutatum, Colletotrichum dematium, and Colletotrichum coccodes. Bacterial isolate GP-P8 from pepper soil was found to be effective against the tested pathogens with an average inhibition rate of 70.7% in in vitro dual culture assays. 16S rRNA gene sequencing analysis result showed that the effective bacterial isolate as Bacillus siamensis. Biochemical characterization of GP-P8 was also performed. According to the results, protease and cellulose, siderophore production, phosphate solubilization, starch hydrolysis, and indole-3-acetic acid production were shown by the GP-P8. Using specific primers, genes involved in the production of antibiotics, such as iturin, fengycin, difficidin, bacilysin, bacillibactin, surfactin, macrolactin, and bacillaene were also detected in B. siamensis GP-P8. Identification and analysis of volatile organic compounds through solid phase microextraction/gas chromatography-mass spectrometry (SPME/GC-MS) revealed that acetoin and 2,3-butanediol were produced by isolate GP-P8. In vivo tests showed that GP-P8 significantly reduced the anthracnose disease caused by C. acutatum, and enhanced the growth of pepper plant. Reverse transcription polymerase chain reaction analysis of pepper fruits revealed that GP-P8 treated pepper plants showed increased expression of immune genes such as CaPR1, CaPR4, CaNPR1, CaMAPK4, CaJA2, and CaERF53. These results strongly suggest that GP-P8 could be a promising biocontrol agent against pepper anthracnose disease and possibly a pepper plant growth-promoting agent.

Chemical Fungicides and Bacillus siamensis H30-3 against Fungal and Oomycete Pathogens Causing Soil-Borne Strawberry Diseases

Bo Reen Park,Hyun Jin Son,Jong Hyeob Park,Eun Soo Kim,Seong Jin Heo,Hae Ree Youn,Young Mo Koo,A Yeong Heo,Hyong Woo Choi,Mee Kyung Sang,Sang-Woo Lee,Sung Hwan Choi,Jeum Kyu Hong 한국식물병리학회 2021 Plant Pathology Journal Vol.37 No.1

Chemical and biological agents were evaluated to in- hibit Colletotrichum fructicola, Phytophthora cactorum, and Lasiodiplodia theobromae causing strawberry dis- eases. Mycelial growths of C. fructicola were gradually arrested by increasing concentrations of fungicides pyraclostrobin and iminoctadine tris (albesilate). P. cac- torum and L. theobromae were more sensitive to pyra- clostrobin compared to C. fructicola, but iminoctadine tris (albesilate) was not or less effective to limit P. cac- torum or L. theobromae, respectively. Bacillus siamensis H30-3 was antagonistic against the three pathogens by diffusible as well as volatile molecules, and evidently reduced aerial mycelial formation of P. cactorum. B. siamensis H30-3 growth was declined by at least 0.025 mg/ml of pyraclostrobin. The two fungicides additively inhibited mycelial growths of C. fructicola, but not of P. cactorum and L. theobromae. B. siamensis H30-3 vola- tiles led to less growth of C. fructicola than one reduced by the fungicides. Taken together, in vitro antimicrobial activities of the two fungicides together with or without B. siamensis H30-3 volatiles may be cautiously incorpo- rated into integrated management of strawberry dis- eases dependent on causal pathogens.

Bacillus siamensis 3BS12-4 Extracellular Compounds as a Potential Biological Control Agent against Aspergillus flavus

Aphaiso Patapee,Mahakhan Polson,Sawaengkaew Jutaporn 한국미생물·생명공학회 2024 Journal of microbiology and biotechnology Vol.34 No.8

Aspergillus flavus, the primary mold that causes food spoilage, poses significant health and economic problems worldwide. Eliminating A. flavus growth is essential to ensure the safety of agricultural products, and extracellular compounds (ECCs) produced by Bacillus spp. have been demonstrated to inhibit the growth of this pathogen. In this study, we aimed to identify microorganisms efficient at inhibiting A. flavus growth and degrading aflatoxin B1. We isolated microorganisms from soil samples using a culture medium containing coumarin (CM medium) as the sole carbon source. Of the 498 isolates grown on CM medium, only 132 bacterial strains were capable of inhibiting A. flavus growth. Isolate 3BS12-4, identified as Bacillus siamensis, exhibited the highest antifungal activity with an inhibition ratio of 43.10%, and was therefore selected for further studies. The inhibition of A. flavus by isolate 3BS12-4 was predominantly attributed to ECCs, with a minimum inhibitory concentration and minimum fungicidal concentration of 0.512 g/ml. SEM analysis revealed that the ECCs disrupted the mycelium of A. flavus. The hydrolytic enzyme activity of the ECCs was assessed by protease, β-1,3-glucanase, and chitinase activity. Our results demonstrate a remarkable 96.11% aflatoxin B1 degradation mediated by ECCs produced by isolate 3BS12-4. Furthermore, treatment with these compounds resulted in a significant 97.93% inhibition of A. flavus growth on peanut seeds. These findings collectively present B. siamensis 3BS12-4 as a promising tool for developing environmentally friendly products to manage aflatoxin-producing fungi and contribute to the enhancement of agricultural product safety and food security.

Peony Stem Rots by Neopestalotiopsis clavispora and Sclerotinia sclerotiorum, and Antifungal Propineb and Bacillus siamensis H30-3 against the Two Fungal Species

Jeum Kyu Hong,Young Hee Lee,Yeon Sook Jo,Su Min Kim,Seoung Bin Lee,Juyeoung Um,Kyoung-Ok Choi,Mee Kyung Sang,Chung-Ryul Jung,Chang-Jin Park,Sung Hwan Choi The Korean Society of Plant Pathology 2024 식물병연구 Vol.30 No.2

In July 2022, stem rot symptom was found in a peony plant grown in a pot under a greenhouse at Jinju, Gyeongnam Province, South Korea. Two fungal species were isolated from the infected peony stems and cultured on 1/2-strength potato dextrose agar for identification. The morphological characteristics of the fungal isolates were examined, and nucleotide sequences of the internal transcribed spacer region, β-tubulin and translation elongation factor 1-α were analysed. The pathogenicity of the two isolates was confirmed in detached peony leaves, according to Koch's postulates. To our knowledge, this is the report of Neopestalotiopsis clavispora and Sclerotinia sclerotiorum as the causal agents of peony stem rots. Antifungal activity of chemical fungicide propineb and rhizobacterium Bacillus siamensis H30-3 was shown against the two plant pathogenic fungi N. clavispora and S. sclerotiorum.Unidentified diffusible and volatile compounds from B. siamensis H30-3 could suppress in vitro mycelial growths of N. clavispora JJ 8-2-1 and S. sclerotiorum JJ 8-2-2.