• 제목/요약/키워드: Microbial Biotechnology

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사람 및 가축 유래 분변 미생물 군집과 항생제 내성 유전자 간 상관 관계에 대한 연구 (Co-occurrence Analyses of Antibiotic Resistance Genes and Microbial Community in Human and Livestock Animal Feces)

  • 정지원;반다리 아프라지타;운노 타쯔야
    • 한국환경농학회지
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    • 제41권4호
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    • pp.335-343
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    • 2022
  • BACKGROUND: Antibiotics used in animal husbandry for disease prevention and treatment have resulted in the rapid progression of antibiotic resistant bacteria which can be introduced into the environment through livestock feces/manure, disseminating antibiotic resistant genes (ARGs). In this study, fecal samples were collected from the livestock farms located in Jeju Island to investigate the relationship between microbial communities and ARGs. METHODS AND RESULTS: Illumina MiSeq sequencing was applied to characterize microbial communities within each fecal sample. Using quantitative PCR (qPCR), ten ARGs encoding tetracycline resistance (tetB, tetM), sulfonamide resistance (sul1, sul2), fluoroquinolone resistance (qnrD, qnrS), fluoroquinolone and aminoglycoside resistance (aac(6')-Ib), beta-lactam resistance (blaTEM, blaCTX-M), macrolide resistance (ermC), a class 1 integronsintegrase gene (intI1), and a class 2 integrons-integrase gene (intI2) were quantified. The results showed that Firmicutes and Bacteroidetes were dominant in human, cow, horse, and pig groups, while Firmicutes and Actinobacteria were dominant in chicken group. Among ARGs, tetM was detected with the highest number of copies, followed by sul1 and sul2. Most of the genera belonging to Firmicutes showed positive correlations with ARGs and integron genes. There were 97, 34, 31, 25, and 22 genera in chicken, cow, pig, human, and horse respectively which showed positive correlations with ARGs and integron genes. In network analysis, we identified diversity of microbial communities which correlated with ARGs and integron genes. CONCLUSION(S): In this study, antibiotic resistance patterns in human and livestock fecal samples were identified. The abundance of ARGs and integron genes detected in the samples were associated with the amount of antibiotics commonly used for human and livestocks. We found diverse microbial communities associated with antibiotics resistance genes in different hosts, suggesting that antibiotics resistance can disseminate across environments through various routes. Identifying the routes of ARG dissemination in the environment would be the first step to overcome the challenge of antibiotic resistance in the future.

Two Bacterial Entophytes Eliciting Both Plant Growth Promotion and Plant Defense on Pepper (Capsicum annuum L.)

  • Kang, Seung-Hoon;Cho, Hyun-Soo;Cheong, Hoon;Ryu Choong-Min;Kim, Ji-Hyun;Park, Seung-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제17권1호
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    • pp.96-103
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    • 2007
  • Plant growth-promoting rhizobacteria (PGPR) have the potential to be used as microbial inoculants to reduce disease incidence and severity and to increase crop yield. Some of the PGPR have been reported to be able to enter plant tissues and establish endophytic populations. Here, we demonstrated an approach to screen bacterial endophytes that have the capacity to promote the growth of pepper seedlings and protect pepper plants against a bacterial pathogen. Initially, out of 150 bacterial isolates collected from healthy stems of peppers cultivated in the Chungcheong and Gyeongsang provinces of Korea, 23 putative endophytic isolates that were considered to be predominating and representative of each pepper sample were selected. By phenotypic characterization and partial 16S rDNA sequence analysis, the isolates were identified as species of Ochrobacterium, Pantoea, Pseudomonas, Sphingomonas, Janthinobacterium, Ralstonia, Arthrobacter, Clavibacter, Sporosarcina, Acidovorax, and Brevundimonas. Among them, two isolates, PS4 and PS27, were selected because they showed consistent colonizing capacity in pepper stems at the levels of $10^6-10^7CFU/g$ tissue, and were found to be most closely related to Pseudomonas rhodesiae and Pantoea ananatis, respectively, by additional analyses of their entire 16S rDNA sequences. Drenching application of the two strains on the pepper seedlings promoted significant growth of peppers, enhancing their root fresh weight by 73.9% and 41.5%, respectively. The two strains also elicited induced systemic resistance of plants against Xanthomonas axonopodis pv. vesicatoria.

Biocatalytic Production of Aldehyde by a Methanol Utilizing Yeast, Hansenula nonfermentans KYP-l Grown in Methanol-limited Continuous Culture

  • Yoon, Byung-Dae;Kim, Hee-Sik;Kwon, Tae-Jong;Yang, Ji-Won;Kwon, Gi-Seok;Lee, Hyun-Sun;Ahn, Jong-Seog;Mheen, Tae-Ick
    • Journal of Microbiology and Biotechnology
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    • 제2권4호
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    • pp.278-283
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    • 1992
  • Aldehyde production by cells of a methanol utilizing yeast, Hansenula nonfermentans KYP-1 was improved when they were grown in a methanol-limited continuous culture, in comparison with cells grown in a batch culture. A higher cell yield was also obtained in continuous culture than in batch culture. This could be due to the fact that a lower methanol concentration was maintained in the jar fermentor to minimize growth inhibition by methanol. A maximum cell productivity of 0.219 g.$liter^{-1}.hr^{-l}$ and a cell yield of 47% were obtained at dilution rates of 0.1 $hr{-1}$ and 0.06 hr{-1}, respectively. The greatest amount of aldehyde was measured at a dilution rate of 0.08 $hr{-1}$. Under optimum reaction conditions, 915.7 mM of acetaldehyde was produced from 1.5 M ethanol after 21 hours reaction, with a conversion rate of 61%. Propionaldehyde and acrolein were produced with conversion rates of 32.7% and 44%, respectively.

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Humic Substances Act as Electron Acceptor and Redox Mediator for Microbial Dissimilatory Azoreduction by Shewanella decolorationis S12

  • Hong, Yi-Guo;Guo, Jun;Xu, Zhi-Cheng;Xu, Mei-Ying;Sun, Guo-Ping
    • Journal of Microbiology and Biotechnology
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    • 제17권3호
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    • pp.428-437
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    • 2007
  • The potential for humic substances to serve as terminal electron acceptors in microbial respiration and the effects of humic substances on microbial azoreduction were investigated. The dissimilatory azoreducing microorganism Shewanella decolorationis S12 was able to conserve energy to support growth from electron transport to humics coupled to the oxidation of various organic substances or $H_2$. Batch experiments suggested that when the concentration of anthraquinone-2-sulfonate (AQS), a humics analog, was lower than 3 mmol/l, azoreduction of strain S12 was accelerated under anaerobic condition. However, there was obvious inhibition to azoreduction when the concentration of the AQS was higher than 5 mmol/l. Another humics analog, anthraquinone-2-sulfonate (AQDS), could still prominently accelerate azoreduction, even when the concentration was up to 12 mmol/l, but the rate of acceleration gradually decreased with the increasing concentration of the AQDS. Toxic experiments revealed that AQS can inhibit growth of strain S12 if the concentration past a critical one, but AQDS had no effect on the metabolism and growth of strain S12 although the concentration was up to 20 mmol/l. These results demonstrated that a low concentration of humic substances not only could serve as the terminal electron acceptors for conserving energy for growth, but also act as redox mediator shuttling electrons for the anaerobic azoreduction by S. decolorationis S12. However, a high concentration of humic substances could inhibit the bacterial azoreduction, resulting on the one hand from the toxic effect on cell metabolism and growth, and on the other hand from competion with azo dyes for electrons as electron acceptor.

Distribution of Toxin Genes and Enterotoxins in Bacillus thuringiensis Isolated from Microbial Insecticide Products

  • Cho, Seung-Hak;Kang, Suk-Ho;Lee, Yea-Eun;Kim, Sung-Jo;Yoo, Young-Bin;Bak, Yeong-Seok;Kim, Jung-Beom
    • Journal of Microbiology and Biotechnology
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    • 제25권12호
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    • pp.2043-2048
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    • 2015
  • Bacillus thuringiensis microbial insecticide products have been applied worldwide. Although a few cases of B. thuringiensis foodborne illness have been reported, little is known about the toxigenic properties of B. thuringiensis isolates. The aims of this study were to estimate the pathogenic potential of B. thuringiensis selected from microbial insecticide products, based on its possession of toxin genes and production of enterotoxins. Fifty-two B. thuringiensis strains selected from four kinds of microbial insecticide products were analyzed. PCR assay for detection of toxin genes and immunoassay for detection of enterotoxins were performed. The hemolysin BL complex as a major enterotoxin was produced by 17 (32.7%), whereas the non-hemolytic enterotoxin complex was detected in 1 (1.9%) of 52 B. thuringiensis strains. However, cytK, entFM, and ces genes were not detected in any of the tested B. thuringiensis strains. The potential risk of food poisoning by B. thuringiensis along with concerns over B. thuringiensis microbial insecticide products has gained attention recently. Thus, microbial insecticide products based on B. thuringiensis should be carefully controlled.

High ${\beta}$-Glucosidase Secretion in Saccharomyces cerevisiae Improves the Efficiency of Cellulase Hydrolysis and Ethanol Production in Simultaneous Saccharification and Fermentation

  • Tang, Hongting;Hou, Jin;Shen, Yu;Xu, Lili;Yang, Hui;Fang, Xu;Bao, Xiaoming
    • Journal of Microbiology and Biotechnology
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    • 제23권11호
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    • pp.1577-1585
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    • 2013
  • Bioethanol production from lignocellulose is considered as a sustainable biofuel supply. However, the low cellulose hydrolysis efficiency limits the cellulosic ethanol production. The cellulase is strongly inhibited by the major end product cellobiose, which can be relieved by the addition of ${\beta}$-glucosidase. In this study, three ${\beta}$-glucosidases from different organisms were respectively expressed in Saccharomyces cerevisiae and the ${\beta}$-glucosidase from Saccharomycopsis fibuligera showed the best activity (5.2 U/ml). The recombinant strain with S. fibuligera ${\beta}$-glucosidase could metabolize cellobiose with a specific growth rate similar to the control strain in glucose. This recombinant strain showed higher hydrolysis efficiency in the cellulose simultaneous saccharification and fermentation, when using the Trichoderma reesei cellulase, which is short of the ${\beta}$-glucosidase activity. The final ethanol concentration was 110% (using Avicel) and 89% (using acid-pretreated corncob) higher than the control strain. These results demonstrated the effect of ${\beta}$-glucosidase secretion in the recombinant S. cerevisiae for enhancing cellulosic ethanol conversion.

Cloning, Expression, and Characterization of a New Phytase from the Phytopathogenic Bacterium Pectobacterium wasabiae DSMZ 18074

  • Shao, Na;Huang, Huoqing;Meng, Kun;Luo, Huiying;Wang, Yaru;Yang, Peilong;Yao, Bin
    • Journal of Microbiology and Biotechnology
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    • 제18권7호
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    • pp.1221-1226
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    • 2008
  • The soft rot bacterium Pectobacterium wasabiae is an economically important pathogen of many crops. A new phytase gene, appA, was cloned from P. wasabiae by degenerate PCR and TAIL-PCR. The open reading frame of appA consisted of 1,302 bp encoding 433 amino acid residues, including 27 residues of a putative signal peptide. The mature protein had a molecular mass of 45 kDa and a theoretical pI of 5.5. The amino acid sequence contained the conserved active site residues RHGXRXP and HDTN of typical histidine acid phosphatases, and showed the highest identity of 48.5% to PhyM from Pseudomonas syringae. The gene fragment encoding the mature phytase was expressed in Escherichia coli BL21 (DE3), and the purified recombinant phytase had a specific activity of 1,072$\pm$47 U/mg for phytate substrate. The optimum pH and temperature for the purified phytase were pH 5.0 and 50$^{\circ}C$, respectively. The $K_m$ value was 0.17 mM, with a $V_{max}$ of 1,714 $\mu$mol/min/mg. This is the first report of the identification and isolation of phytase from Pectobacterium.

막걸리 유래 미생물의 활용을 위한 연구 동향 (Application of Functional Microbial Strains Isolated from Traditional Rice Wine in Korea)

  • 이영석;설정만;정덕열;김수린
    • 한국미생물·생명공학회지
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    • 제44권3호
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    • pp.229-235
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    • 2016
  • 우리나라는 예전부터 전통적인 방법으로 누룩을 제조하고, 이를 발효제로 막걸리를 만들어 왔다. 막걸리는 여과 또는 살균 과정없이 다양한 미생물을 살아 있는 상태로 섭취하기 때문에 영양학적으로나 기능적인 측면에서 가치가 높다. 최근 많은 연구에서 막걸리로부터 미생물을 분리동정하고 다양한 기능성에 대해 스크리닝한 결과, 높은 프로바이오틱스 활성과 다양한 스펙트럼의 항균활성을 가진 균주들이 선별되었다. 특히 일부 유산균들은 GABA와 EPS 등의 기능성 물질을 생성하기도 했다. 또한, 일부 유산균과 효모의 경우 각각 bacteriocin 및 killer toxin을 통해 항균활성을 나타내는 것으로 여겨진다. 이러한 막걸리 유래 기능성 미생물과 그 대사산물은 기능성 식품 소재나 안전한 식품첨가물 및 다양한 산업분야에서 활용될 수 있을 것이다.

Antimicrobial Effect of Calcium Chloride Alone and Combined with Lactic Acid Injected into Chicken Breast Meat

  • Alahakoon, Amali U.;Jayasena, Dinesh D.;Jung, Samooel;Kim, Hyun Joo;Kim, Sun Hyo;Jo, Cheorun
    • 한국축산식품학회지
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    • 제34권2호
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    • pp.221-229
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    • 2014
  • Chicken breast meat was injected with calcium chloride alone and in combination with lactic acid (0.01% and 0.002%, respectively). The inhibitory effects of the treatments on microbial growth were determined in the injected chicken breast meat stored at $4^{\circ}C$ under aerobic packaging condition for 0, 3, and 7 d. Calcium chloride combined with 0.002% and 0.01% lactic acid reduced microbial counts by 0.14 and 1.08 Log CFU/g, respectively, however, calcium chloride alone was unable to inhibit microbial growth. Calcium chloride combined with 0.01% lactic acid was the most effective antimicrobial treatment and resulted in the highest initial redness value. Calcium chloride alone and combined with lactic acid suppressed changes in pH and the Hunter color values during storage. However, injection of calcium chloride and lactic acid had adverse effects on lipid oxidation and sensory characteristics. The higher TBARS values were observed in samples treated with calcium chloride and lactic acid when compared to control over the storage period. Addition of calcium chloride and lactic acid resulted in lower sensory scores for parameters tested, except odor and color, compared to control samples. Therefore, the formulation should be improved in order to overcome such defects prior to industrial application.

HaCaT 세포주에서 캐모마일 (Matricaria chamomilla) 추출물의 항병원 성 및 항염 효과에 관한 연구 (Study of Anti-microbial Activities and Anti-inflammatory Effects of Chamomile (Matricaria chamomilla) Extracts in HaCaT cells)

  • 임은경;김근태;김보민;김은지;김상용;한남규;하재순;김영민
    • KSBB Journal
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    • 제32권1호
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    • pp.9-15
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    • 2017
  • Chamomile (Matricaria chamomilla), a member of the Asteraceae family, is a well-known for medicinal plant and can be found in India and Europe. Chamomile is an effective sedative and various medical effects. But, the effects of acne treatment by chamomile were not investigated. Therefore, we assessed the anti-oxidant effects, anti-microbial activity and anti-inflammatory effects by chamomile extracts in HaCaT keratinocyte cells. Anti-oxidant effects of chamomile extracts were investigated by DPPH assay. Also, results of MTT assay was demonstrated that chamomile extracts did not have a cytotoxic effect in HaCaT cells. To assess the antimicrobial activity, we determined formation of inhibition zone of Propionibacterium acnes by extracts from chamomile. Tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) induces production of inflammatory cytokines such as interleukin-$1{\beta}$ (IL-$1{\beta}$), IL-6 and IL-8 and expression of COX-2. Chamomile extracts could inhibit TNF-${\alpha}$-induced mRNA expression levels of IL-$1{\beta}$, IL-6, IL-8 and COX-2 gene. These results demonstrated the possibility of chamomile for prevention and treatment of skin inflammatory diseases such as acne.