• 제목/요약/키워드: Bacterial culture

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Modification of Acetobacter xylinum Bacterial Cellulose Using Dextransucrase and Alternansucrase

  • Kim, Do-Man;Kim, Young-Min;Park, Mi-Ran;Park, Don-Hee
    • Journal of Microbiology and Biotechnology
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    • 제9권6호
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    • pp.704-708
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    • 1999
  • In addition to catalyzing the synthesis of glucan from sucrose as a primary reaction, glucansucrase also catalyzes the transfer of glucose from sucrose to other carbohydrates that are present or are added to the reaction digest. Using dextransucrase and altemansucrase, prepared from Leuconostoc mesenteroides B-742CBM and B-1355C, respectively, we modified the bacterial cellulose in Acetobacter xylinum ATCC10821 culture, and then produced a characteristic cellulose that is soluble and has a new structure. There were also some partially modified insoluble cellulose and oligosaccharides in the modification culture. After methylation and following acid hydrolysis of both the soluble and insoluble glucans, there were ($1{\rightarrow}4$) as well as ($1{\rightarrow}6$) and ($1{\rightarrow}3$) glycosidic linkages in the soluble glucan.

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온실재배 토마토에서 발생하는 위조병의 미생물학적 제어 (Biological Control of Fusarium Wilt of Tomato Plants by Antagonistic Microorganism in Greenhouse)

  • 조정일;조자용
    • 한국유기농업학회지
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    • 제11권4호
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    • pp.61-74
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    • 2003
  • This study was conducted to screen the antagonistic bacteria which inhibit the growth of plant pathogen, fusarium wilt(Fusarium oxysporum) occurred in tomato plants in greenhouse. We isolated an effective bacterial strains and investigated into the antifungal activity of the antagonistic microorganism and it’s identification. Ten bacterial strains which strongly inhibited Fusarium oxysporum were isolated from the nature, and the best antagonistic bacterial strain designated as KC175, was selected. The antagonistic strain KC175 was identified to be the genus Bacillus sp. based on the morphological and biochemical characterization. The Bacillus sp. KC175 showed 58.2% of antifungal activity against the growth of Fusarium oxysporum. By the bacterialization of the culture broth and the heat bacterialization culture filtrate of it, Bacillus sp. KC175 showed 91% and 18% of antifungal activity, respectively.

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DGGE 방법과 Pyrosequencing 방법을 이용한 지렁이 장내미생물의 다양성 분석 (Comparative Analysis of Bacterial Diversity in the Intestinal Tract of Earthworm (Eisenia fetida) using DGGE and Pyrosequencing)

  • 김은성;홍성욱;정건섭
    • 한국미생물·생명공학회지
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    • 제39권4호
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    • pp.374-381
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    • 2011
  • 미생물과의 상호작용을 통하여 토양의 특성을 변화시킬 수 있는 지렁이 Eisenia fetida의 장내미생물 군집을 조사하기 위하여, 배양방법과 비배양방법인 DGGE와 pyrosequencing을 이용하여 8주와 16주 사육 지렁이의 장내미생물 군집을 분석하였다. 배양방법에서는 L. fusiformis(51%), B. cereus(30%), E. aerogenes(21%), 그리고 L. sphaericus (15%) 등이 우점미생물로 확인되었다. DGGE 분석에서는 B. cereus(15.1%), Enterobacter sp.(13.6%), uncultured bacterium (13.1%), 그리고 B. stearothermophilus(7.8%)가 우점미생물로 확인되었다. Pyrosequencing 분석에서는 Microbacterium soli(26%), B. cereus(10%), M. esteraromaticum(6%), 그리고 Frigoribacterium sp.(6%)가 우점미생물로 확인되었다. 그외에도 Aeromonas sp., Pseudomonas sp., Borrelia sp., Cellulosimicrobium sp., Klebsiella sp., and Leifsonia sp. 등의 미생물도 확인이 되었으며, 비배양 방법을 이용한 장내 미생물 군집 조사는 배양이 불가능한 미생물을 확인할 수 있을 뿐만 아니라 더 다양한 미생물 군집도 확인할 수 있었다.

Interactions between Entodinium caudatum and an amino acid-fermenting bacterial consortium: fermentation characteristics and protozoal population in vitro

  • Tansol Park;Zhongtang Yu
    • Journal of Animal Science and Technology
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    • 제65권2호
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    • pp.387-400
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    • 2023
  • Ruminal protozoa, especially entodiniomorphs, engulf other members of the rumen microbiome in large numbers; and they release oligopeptides and amino acids, which can be fermented to ammonia and volatile fatty acids (VFAs) by amino acid-fermenting bacteria (AAFB). Studies using defaunated (protozoa-free) sheep have demonstrated that ruminal protozoa considerably increase intraruminal nitrogen recycling but decrease nitrogen utilization efficiency in ruminants. However, direct interactions between ruminal protozoa and AAFB have not been demonstrated because of their inability to establish axenic cultures of any ruminal protozoan. Thus, this study was performed to evaluate the interaction between Entodinium caudatum, which is the most predominant rumen ciliate species, and an AAFB consortium in terms of feed degradation and ammonia production along with the microbial population shift of select bacterial species (Prevotella ruminicola, Clostridium aminophilum, and Peptostreptococcus anaerobius). From an Ent. caudatum culture that had been maintained by daily feeding and transfers every 3 or 4 days, the bacteria and methanogens loosely associated with Ent. caudatum cells were removed by filtration and washing. An AAFB consortium was established by repeated transfers and enrichment with casamino acids as the sole substrate. The cultures of Ent. caudatum alone (Ec) and AAFB alone (AAFB) and the co-culture of Ent. caudatum and AAFB (Ec + AAFB) were set up in three replicates and incubated at 39℃ for 72 h. The digestibility of dry matter (DM) and fiber (NDF), VFA profiles, ammonia concentrations, pH, and microscopic counts of Ent. caudatum were compared among the three cultures. The co-culture of AAFB and Ent. caudatum enhanced DM degradation, VFA production, and Ent. caudatum cell counts; conversely, it decreased acetate: propionate ratio although the total bacterial abundance was similar between Ec and the Ec + AAFB co-culture after 24 h incubation. The ammonia production and relative abundance of C. aminophilum and P. anaerobius did not differ between AAFB alone and the Ec + AAFB co-culture. Our results indicate that Ent. caudatum and AAFB could have a mutualistic interaction that benefited each other, but their interactions were complex and might not increase ammoniagenesis. Further research should examine how such interactions affect the population dynamics of AAFB.

A report of 44 unrecorded bacterial species isolated from Nakdong River in Korea

  • Ju-Hyung Jeon;Sanghwa Park;Ja Young Cho;Soo-Yeong Lee;Seoni Hwang;Jun Sung Kim;Eui-Jin Kim ; Ji Young Jung
    • 환경생물
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    • 제41권3호
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    • pp.308-324
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    • 2023
  • This study investigated unrecorded freshwater bacterial species in Korea. Water and sediment samples were collected from the Nakdong River basin from 2020-2022. Bacterial isolates obtained through the conventional culture method with commercial media were subjected to 16S rRNA gene sequencing to identify unrecorded bacterial species. Results of 16S rRNA gene sequencing of the bacterial isolates revealed that a total of 44 bacterial isolates shared 16S rRNA gene sequence similarities of more than 98.65%, with validly published bacterial species not reported in Korea yet. These isolates were phylogenetically assigned to 4 phyla, 7 classes, 21 orders, 33 families, and 42 genera. A total of 2, 6, 12, and 24 species belonged to phyla Bacillota, Bacteroidota, Actinomycetota, and Pseudomonadota, respectively. Here, we provide details of these 44 unrecorded bacterial species, including Gram staining, colony and cellular morphologies, biochemical properties, and phylogenetic position.

Changes in the ruminal fermentation and bacterial community structure by a sudden change to a high-concentrate diet in Korean domestic ruminants

  • Lee, Mingyung;Jeong, Sinyong;Seo, Jakyeom;Seo, Seongwon
    • Asian-Australasian Journal of Animal Sciences
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    • 제32권1호
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    • pp.92-102
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    • 2019
  • Objective: To investigate changes in rumen fermentation characteristics and bacterial community by a sudden change to a high concentrate diet (HC) in Korean domestic ruminants. Methods: Major Korean domestic ruminants (each of four Hanwoo cows; $545.5{\pm}33.6kg$, Holstein cows; $516.3{\pm}42.7kg$, and Korean native goats; $19.1{\pm}1.4kg$) were used in this experiment. They were housed individually and were fed ad libitum with a same TMR (800 g/kg timothy hay and 200 g/kg concentrate mix) twice daily. After two-week feeding, only the concentrate mix was offered for one week in order to induce rapid rumen acidosis. The rumen fluid was collected from each animals twice (on week 2 and week 3) at 2 h after morning feeding using an oral stomach tube. Each collected rumen fluid was analyzed for pH, volatile fatty acid (VFA), and $NH_3-N$. In addition, differences in microbial community among ruminant species and between normal and an acidosis condition were assessed using two culture-independent 16S polymerase chain reaction (PCR)-based techniques (terminal restriction fragment length polymorphism and quantitative real-time PCR). Results: The HC decreased ruminal pH and altered relative concentrations of ruminal VFA (p<0.01). Total VFA concentration increased in Holstein cows only (p<0.01). Terminal restriction fragment length polymorphism and real-time quantitative PCR analysis using culture-independent 16S PCR-based techniques, revealed rumen bacterial diversity differed by species but not by HC (p<0.01); bacterial diversity was higher in Korean native goats than that in Holstein cows. HC changed the relative populations of rumen bacterial species. Specifically, the abundance of Fibrobacter succinogenes was decreased while Lactobacillus spp. and Megasphaera elsdenii were increased (p<0.01). Conclusion: The HC altered the relative populations, but not diversity, of the ruminal bacterial community, which differed by ruminant species.

볏짚 조사료에 대한 효모 배양물 첨가가 반추위 소화율 및 섬유소 분해균의 군락 변화에 미치는 영향 (Effects of Yeast Culture Supplementation on Rice Straw Digestibility and Cellulolytic Bacterial Community in the Rumen)

  • 성하균
    • Journal of Animal Science and Technology
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    • 제55권1호
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    • pp.41-49
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    • 2013
  • 본 연구는 효모(Saccharomyces cerevisiae)가 조사료의 반추위 미생물 분해 및 섬유소 분해 박테리아의 군집 변화에 미치는 관계를 검증하고자, 생효모 배양물 첨가 및 급여가 볏짚 소화율에 미치는 영향을 in vitro 및 in situ 실험을 통해 측정하였고, 볏짚 분해 시 볏짚 표면에 부착한 섬유소분해 박테리아 군집의 변화를 real-time PCR 방법을 이용하여 비교 분석하였다. 생효모 배양물의 첨가 수준(0.0, 0.2, 0.4, 0.6, 0.8 및 1.0%)에 따른 볏짚의 in vitro 건물 소화율을 비교 하였을 때 첨가 수준이 증가 할수록 소화율도 점진적으로 높아 졌다. 특히, 생효모 배양물의 첨가 수준을 0.6% 이상으로 하였을 때 0.0, 0.2 및 0.4% 첨가보다도 확연한 소화율 증가를 보이기 시작하였다(p<0.05). 또한 효모배양물의 0.6% 첨가를 NaOH 4% 처리 볏짚 및 무처리 볏짚에 적용하여 소화율을 재평가하였을 때 두 가지 볏짚 모두에서 효모배양물 첨가에 의하여 볏짚 소화율이 유의적으로 증가하였다(p<0.05). 생효모 배양물을 한국재래산양에 실제 급여에 의한 볏짚의 in situ 건물소화율을 24 및 48 시간 후에 관측하였을 때 NaOH 무처리 볏짚의 효모배양물 비급여구의 소화율이 가장 낮았고, NaOH 무처리 볏짚의 효모배양물 급여구, 4% NaOH 처리 볏짚의 효모배양물 비급여구, 그리고 4% NaOH 처리 볏짚의 효모배양물 급여구 순으로 소화율이 유의적으로 각각 증가하였다(p<0.05). 이는 NaOH 무처리 볏짚 및 4% NaOH 처리 볏짚 모두에서 생효모 배양물 급여에 의하여 in situ 건물 소화율이 유의적으로 증가함을 보여준 것이다(p<0.05). 그리고 볏짚을 효모배양물의 두 가지 농도로 볏짚을 분무 처리하여 in situ 소화율과 볏짚 표면 부착 섬유소분해 박테리아(F. succiongenes, R. flavefaciens, R. albus)의 군집 변화를 측정하였을 때 효모배양물 처리 농도가 증가함에 따라 소화율도 유의적으로 높게 나타났고(p<0.05), 동시에 이들 박테리아의 볏짚 표면 부착 군집도 효모배양물의 처리 농도 증가에 따라 증가 하였다. F. succiongenes과 R. flavefaciens는 배양 12 및 24시간 모두 처리농도에 따라 군락의 수가 유의적으로 증가하였다(p<0.05). 그리고 R. albus도 배양 12 시간 처리농도에 따라 군락의 수가 유의적으로 증가하였고(p<0.05), 24 시간도 처리농도에 따라 증가 경향을 나타냈다. 따라서 본 연구는 효모 첨가는 조사료 소화율을 증진에 확실히 좋은 영향을 주며, 이것은 조사료 표면 부착 섬유소분해 박테리아의 군락 형성 증가에서 기인함을 보여준다고 사료된다.

Different Profiles of the Negatively Stained Citrus Canker Bacterium Xanthomonas citri pv. citri Depending on Culture Media and Heavy Metal Stains

  • Kim, Ki-Woo;Lee, In-Jung;Hyun, Jae-Wook;Lee, Yong-Hoon;Park, Eun-Woo
    • The Plant Pathology Journal
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    • 제26권1호
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    • pp.90-92
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    • 2010
  • Staining profiles and bacterial morphology were compared in Xanthomonas citri pv. citri by a transmission electron microscopy. Four types of negative staining regimes were employed depending on culture media and heavy metal stains. The bacterial cells grown on LB agar media often appeared clustered on the supporting film. Meanwhile, individual bacterial cells could be readily found on the preparations from LB broth media. Typical rod-shaped cells (ca. $1\;{\mu}m$ in length) and their flagella were observed in either 2% uranyl acetate (UA) or 2% neutralized potassium phosphotungstate (PTA) staining. The UA-stained bacteria often showed relatively intact cell morphology and rather positively stained cells with a thin electron-dense stain depth around bacteria. The PTA-stained bacteria were characterized by the wrinkled cell surface where the stain was entrapped in grooves. In addition, distinct electron-dense stain depth was evident around the PTA-stained preparations. Numerous fimbriae could be mostly observed from the PTA-stained preparations of the two culture media, but not from the UA-stained preparations.

Isolation of a Lipolytic and Proteolytic Bacillus licheniformis from Refinery Oily Sludge and Optimization of Culture Conditions for Production of the Enzymes

  • Devi, Sashi Prava;Jha, Dhruva Kumar
    • 한국미생물·생명공학회지
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    • 제48권4호
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    • pp.515-524
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    • 2020
  • With the increasing demand for enzymes in industrial applications there is a growing need to easily produce industrially important microbial enzymes. This study was carried out to screen the indigenous refinery bacterial isolates for their production of two industrially important enzymes i.e. lipase and protease. A total of 15 bacterial strains were isolated using Soil Extract Agar media from the oil-contaminated environment and one was shown to produce high quality lipase and protease enzymes. The culture conditions (culture duration, temperature, source of nitrogen, carbon, and pH) were optimized to produce the optimum amount of both the lipase (37.6 ± 0.2 Uml-1) and the protease (41 ± 0.4 Uml-1) from this isolate. Productivity of both enzymes was shown to be maximized at pH 7.5 in a medium containing yeast extract and peptone as nitrogen sources and sucrose and galactose as carbon sources when incubated at 35 ± 1℃ for 48 h. Bacterial strain SAB06 was identified as Bacillus licheniformis (MT250345) based on biochemical, morphological, and molecular characteristics. Further studies are required to evaluate and optimize the purification and characterization of these enzymes before they can be recommended for industrial or environmental applications.

활성탄 및 세라믹 재질에의 세균 부착성에 대한 연구 (An Experimental Study on Bacterial Adhesion onto Activated Carbon and Ceramic)

  • 권성현;조대철;이인형
    • 한국환경과학회지
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    • 제14권12호
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    • pp.1163-1170
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    • 2005
  • The microbial adsorption characteristics of two different media for biological treatment were studied using attached diverse microbes onto activated carbon and ceramic. The results in the experiments of the characteristics of physical adhesion on two different media with addition of high and low concentrated substrate in the culture were observed that the efficient of adhesion onto F-400 activated carbon was higher over that of ceramic due to the surface area of media. The irradiation treatment by ultrasonication with 400 W power and 3 min retention time on the media without addition substrate conditions and subsequent mixing throughly the culture showed the highest efficiency of cell detachment on the media. Three different microbes, P. ovalis, A calcoaceticus, and B. subtillis were used for the study of the characteristics of microbial adhesion on the media. p ovalis showed the highest adhesion capability while B. subtillis showed the lowest capability adhesion onto media either addition of substrate in the culture. The mixed bacterial culture showed $10\%$ lower removal efficiency of DOC in the low concentrated substrate culture compared to the single pure culture. Whileas, it did not show significant difference between two cultures at high concentrated substrate. It was also observed same population density of microorganism by counting of microbes adhered to microbial media with an ultrasound treatment.