• Title/Summary/Keyword: bacterial production

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Production of a hypothetical polyene substance by activating a cryptic fungal PKS-NRPS hybrid gene in Monascus purpureus (홍국Monascus purpureus에서 진균 PKS-NRPS 하이브리드 유전자의 발현 유도를 통한 미지 polyene 화합물의 생성)

  • Suh, Jae-Won;Balakrishnan, Bijinu;Lim, Yoon Ji;Lee, Doh Won;Choi, Jeong Ju;Park, Si-Hyung;Kwon, Hyung-Jin
    • Journal of Applied Biological Chemistry
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    • v.61 no.1
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    • pp.83-91
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    • 2018
  • Advances in bacterial and fungal genome mining uncover a plethora of cryptic secondary metabolite biosynthetic gene clusters. Guided by the genome information, targeted transcriptional derepression could be employed to determine the product of a cryptic gene cluster and to explore its biological role. Monascus spp. are food grade filamentous fungi popular in eastern Asia and several genome data belong to them are now available. We achieved transcription activation of a cryptic fungal polyketide synthase-nonribosomal peptide synthase gene Mpfus1 in Monascus purpureus ${\Delta}MpPKS5$ by inserting Aspergillus gpdA promoter at the upstream of Mpfus1 through double crossover gene replacement. The gene cluster with Mpfus1 show a high similarity to those for the biosynthesis of conjugated polyene derivatives with 2-pyrrolidone ring and the mycotoxin fusarin is the representative member of this group. The ${\Delta}MpPKS5$ is incapable of producing azaphilone pigment, providing an excellent background to identify chromogenic and UV-absorbing compounds. Activation of Mpfus1 resulted in a yellow hue on mycelia and its methanol extract exhibit a maximum absorption at 365 nm. HPLC analysis of the organic extracts indicated the presence of a variety of yellow compounds in the extract. This implies that the product of MpFus1 is metabolically or chemically unstable. LC-MS analysis guided us to predict the MpFus1 product and to propose that the Mpfus1-containing gene cluster encode the biosynthesis of a desmethyl analogue of fusarin. This study showcases the genome mining in Monascus and the possibility to unveil new biological activities embedded in it.

Microbiological and Sensory Characteristics of Vacuum Packed Korean Chilled Pork Loins for Export (한국산 수출용 진공포장 냉장 돈육 등심의 미생물학적 및 관능적 품질특성)

  • Choi, Y.S.;Park, B.Y.;Lee, J.M.;Kim, I.S.;Lee, S.K.;Kim, B.C.
    • Journal of Animal Science and Technology
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    • v.44 no.3
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    • pp.351-360
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    • 2002
  • The Korean fresh pork loins in vacuum packaged were obtained from three different Korean export companies and investigated for microbiological and sensory properties. The fresh pork loins were stored at 2$^{\circ}C$ for 50 days and analyzed with an interval of 5$\sim$10 days. The results were as follows: The overall numbers of total plate counts and coliform bacteria were higher in swab method than in meat sampling method. The total plate counts in the loins from the company I were maintained low levels ($\prec$10$^5$ cfu/$cm^2$ or $\prec$10$^5$ cfu/g) for entire storage periods(50 days at 2$^{\circ}C$), whereas the loins from the company III had high levels when they were compared to the domestic standard for the allowance limit. The samples from the company III showed that total plate counts were over 106 after about 30 days when determined by meat sampling method and total plate counts were over 106 after 15 days when determined by swab method. The overall numbers of coliform bacteria were also significantly lowest in the samples from the company I, whereas they were highest in the company III. Therefore, all meat companies will have to make an effort to prevent bacterial contamination in each stage such as slaughtering, marketing and consumer in order to ensure the production of safe meat and the extension of shelf-life. For fresh products, scores of intramuscular fat were higher in samples form the companies II and III than those from the company I when visibly evaluated with the standard. There were no significant differences in scores of meat color, drip and fresh meat flavor. However, the samples from the company I had the lowest score of off-flavor and highest score of overall acceptability. For cooked products, there were no significant differences in meat flavor, off-flavor, juiciness and overall acceptability.

A Domestic Outbreak of Bacterial Dysentery Caused by Extended-Spectrum β-Lactamase(ESBL)-producing Shigella sonnei (집단 발생한 Extended-spectrum β-lactamase(ESBL) 생산 Shigella sonnei 감염에 의한 세균성 이질에 관한 연구)

  • Lim, Hyun Taek;Lee, So Hee;Lee, Jung Hwa;Kim, Jeong Eun;Kim, Kyo Sun;Jeong, Eun Ju;Lee, Seung Hyun;Kang, Chang Gyu;Hong, Seong Jin
    • Clinical and Experimental Pediatrics
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    • v.48 no.10
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    • pp.1107-1115
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    • 2005
  • Purpose : An outbreak of ESBL-producing Shigella sonnei enteritis was unprecedented not only in Korea but throughout the world in the past. We intended to devise a management guideline for ESBL-producing shigellosis based on analysis of clinical manifestations and response to therapy. Methods : We analyzed 103 patients who were admitted to the hospital with acute GI symptoms and were shown positive result for S. sonnei on stool culture. We performed sensitivity test to the antibiotics and DNA sequencing of ESBL gene in the isolated S. sonnei colonies. In addition, we retrospectively analyzed their clinical characteristics, laboratory results, and clinical and microbiological responses to the antibiotics. Results : Among the clinical manifestations, fever was the most frequent(96.1%), followed by diarrhea(93.2%), abdominal pain(76.7%), headache(71.8%), vomiting(65.0%), and nausea(41.7%). The fever was sustained for average of 2.0 days and diarrhea for 3.9 days. Watery diarrhea was the most common(69%) followed by mucoid(26%), and bloody stool(5%). On peripheral blood smear, leukocytosis was noted in 53.4% of patients, and 78.6% of patients tested positive for serum CRP response. On stool direct smear, 11.7% of patients showed more than 50 WBCs/HPF, and 9.7% of patients between 5 to 20 WBCs/HPF. Stool occult blood was positive in 71% of patients. Production of CTX-M-14 type ESBL was reported for all S. sonnei strains isolated from this outbreak. Microbiological eradication rates to various antibiotics were as follows : 100%(9/9) to ciprofloxacin, 100% 5/5) to azithromycin, 6.9%(5/72) to cefdinir, 0%(0/8) to ceftriaxone, 12.5%(1/8) to ceftizoxime, 0%(0/ 8) to TMP/SMX, 42.9%(3/7) to ampicillin/sulbactam, 20%(1/5) to amoxicillin/clavulanic acid, and 68.8 %(11/16) to imipenem/cilastatin. Conclusion : It is presumed that azithromycin can be an attractive option for the treatment of ESBL-producing S. sonnei enteritis in pediatric population, given its cost-effectiveness and safety. Although ciprofloxacin is another cost-effective agent, its use in pediatric population may be a bit too premature.

Immunomodulatory effect of the water extract of Aster tataricus through mitogen-activated protein kinase signaling pathway (Aster tataricus 물 추출물의 mitogen-activated protein kinase 신호 전달 경로를 통한 면역 조절 효과)

  • Lee, Chea Yeon;Park, Hyo Sung;Kong, Deok-Hoon;Kim, Young Kwan;Cho, Whajung
    • Journal of Nutrition and Health
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    • v.53 no.5
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    • pp.452-463
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    • 2020
  • Purpose: Aster tataricus (AT) is one of the Asteraceae perennial herbs used in traditional Chinese medicine. The herb contains various bioactive substances, such as flavonoids, isoflavonoids, and phenolic compounds in the roots, and exhibits a range of effects including anti-bacterial, anti-oxidant, and anti-inflammatory activities. This study compared the immunomodulatory effects of ethanol and water extracts of whole AT, except the roots, and analyzed the molecular mechanisms for the regulatory effects on cytokine secretion from THP-1 cells. Methods: The effects of AT extract on the cell viability and proliferation of THP-1 cells were analyzed using the Cell Counting Kit-8 method. The concentrations of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in the cell culture supernatant of the AT-treated THP-1 cells were measured using an enzyme-linked immunosorbent assay. The protein levels of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), inhibitor of nuclear factor kappa B (IκBα), and mitogen-activated protein kinase (MAPK) phosphorylation in the cell lysates were determined by western blotting. Results: The water extract and the ethanol extract of AT did not affect the cell viability, and increased the proliferation of THP-1 cells significantly compared to the vehicle. The water extract increased the secretion of IL-1β from THP-1 cells in a dose-dependent manner, but the ethanol extract had no effect. The expression of COX-2 and iNOS protein and the phosphorylation of MAPK and Akt were induced in AT-treated cells. In addition, IκBα was degraded by AT in a concentration-dependent manner. IL-1β secretion by AT was reduced by extracellular-signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) inhibitors, while TNF-α secretion was decreased by inhibitors of ERK, p38 MAPK, and JNK. Interestingly, the p38 MAPK inhibitor increased the production of IL-1β by AT further. Conclusion: The water extract of the above-ground parts of AT contains immunomodulatory bioactive substances that stimulate immune cells through the MAPK signaling pathway.

Effects of Cover Plants on Soil Microbial Community in Organic Apple Orchards (피복작물이 유기 사과과원 토양미생물상에 미치는 영향)

  • Oh, Young-Ju;Kang, Seok-Boem;Song, Yang-Ik;Choi, Jin-Ho;Paik, Woen-Ki
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.5
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    • pp.822-828
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    • 2012
  • Organic fruit production has increased due to consumer's interest and government's political support for environmentally-friendly agriculture. The aim of this study was to investigate the effects of cover plants on soil microbial community and establish the fruit cultivation method by organic farming techniques. Cover plants used as an organic nutrient source in an apple orchard were rye and barley, the Gramineae and red clover and hairy vetch, the Leguminosae. In the effects of cover plants on the soil chemical characteristics, the soil pH values were higher than that of conventional organic pear orchard. The content of P showed no significant difference between control and cover plant plots. Organic matter level was similar in control and Gramineae cover plant plots, while organic matter content in cover plants belong to Leguminosae was lower than that of control plot. K content was lower in the plots treated with rye and red clover than control plot, while K content in hairy vetch treated plot was higher than control plot. Ca content was lower in control plot than in cover plant treated plots. Concentrations of Mg in the plots treated with barley and hairy vetch was lower than control plot. In August rye and red clover covered soil showed higher bacterial community density than that of control soil and barley treated soil showed highest Actinomycetes community density among treatments. Barley and hairy vetch soils showed higher level of fungi community density than that of control soil in August. In pyrosequencing analysis barley treated soil showed highest distribution ratio of Actinomycetes among treatment. Our findings might be used as basic data for choosing cover plant with effective organic matter decomposition and nutrition supply capacity.

Elimination and Utilization of Pollutants - Part I Microbiological Clarification of Industrial Waste and Its Utilization as Feed Resources - (환경오염원(環境汚染源)의 제거(除去)와 그 이용성(利用性)에 관(關)한 연구(硏究) - 제(報I)1보(第). 미생물(微生物)에 의(依)한 산업폐수(産業廢水)의 정화(淨化) 및 사료자원개발(飼料資源開發)에 개(開)하여 -)

  • Lee, Ke-Ho;Lee, Kang-Heup;Park, Sung-O
    • Applied Biological Chemistry
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    • v.23 no.1
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    • pp.64-72
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    • 1980
  • Industrial wastes from pulp and food plants were treated with microorganisms to clarify organic waste-water and to produce cells as animal feed, and results were summarized as follows. (1) Waste-water from pulp, beer, bread yeast, and ethanol distillation plants contained $1.4{\sim}1.5%$ of total sugar, $0.25{\sim}0.35%$ nitrogen, and biological oxygen demand (BOD) was $400{\sim}25,000$, chemical oxygen demand (COD), $500{\sim}28,000$, and pH, $3.8{\sim}7.0$. The BOD and COD were highest in waste-water from ethanol distillation plants among others. (2) Bacterial and yeast counts were $4{\times}10^4-1{\times}10^9,\;2{\times}10^2-7{\times}10^4/ml$ in waste-water. (3) Bacteria grew better in pulp waste and yeasts in beer, bread yeast, and ethanol distillation waste. (4) Saccharomyces cerevisiae SAFM 1008 and Candida curvata SAFM 70 were the most suitable microorganisms for clarification of ethanol distillation waste. (5) When liquid and solid waste from ethanol distillation were treated with microbial cellulase, xylanase, and pectinase, solid waste was reduced by 36%, soluble waste was increased, and recuding sugar content was increased by 1.3 times which provided better medium than untreated waste for cultivation of yeasts. (6) Optimum growth conditions of the two species of yeast in ethanol distillation waste were pH 5.0, $30^{\circ}C$, and addition of 0.2% of urea, 0.1% of $KH_2PO_4$ and 0.02% of $MgSO_4$. (7) Minimum number of yeast for proper propagation was $1.8{\times}10^5/ml$. (8) C. curvata70 was better than cerevisae for the production of yeast cells from ethanol distillation waste treated with microbial enzymes. (9) S. cerevisiae produced 16 g of dried cell per 1,000ml of ethanol distillation waste and reduced BOD by 46%. C. curvata produced 17.6g of dried cell and reduced BOD by 52% at the same condition. (10) Yeast cells produced from the ethanol distillation waste contained 46-52% protein indicating suitability as a protein source for animal feed.

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Identification of Novel Psychrotolerant Bacterial Strain and Production of $\beta-Galactosidase$ (새로운 저온 내성세균의 동정과 $\beta-Galactosidase$ 생산)

  • Park, Jeong-Woon;Yoo, Jae-Soo;Roh, Dong-Hyun
    • Korean Journal of Microbiology
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    • v.42 no.1
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    • pp.40-46
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    • 2006
  • Galactose joined to glucose by a $\beta(1\rightarrow4)$ glycosidic bond makes lactose and this disaccharide is rich in milk. It is known that lacotse is hydrolyzed to each monomeric sugar by either lactase in human or $\beta-galactosidase$ in bacteria. Ingestion of milk by lactase-deficient persons causes a temporary diarrhea and subsequent chronic diarrhea results in colitis with chronic inflammation. We isolated a $\beta-galactosidase$ producing psycrotolerant strain AS-20 from near cattle shed and investigated the growth at various temperature conditions. Whereas Escherichia coli strains did not grow at $10^{\circ}C$, the AS-20 strain could grow well at this low temperature and showed optimal growth at $30^{\circ}C$. The isolated strain was identified as 97% Hafnia alvei by biochemical properties. This strain could ferment glucose, lacotse, maltose, mannitol, xylose, ONPG, rhamanose and L-arabinose, and decarboxylate lysin and ornithine. To confirm the identity of isolated strain we amplified 16S rDNA by PCR and searched similarity of the 1426 bp DNA sequcence with Genbank database. The strain AS-20 showed 99% similarity with Hafnia alvei. The activity of $\beta-galactosidase$ was 1.5 times higher when the cell was grown at 10 or $20^{\circ}C$ than at $30^{\circ}C$. The highest enzyme activity of AS-20 was also much higher than that of E. coli, which was grown at $30^{\circ}C$.

Physicochemical Characteristics of Seafood-Added Kimchi during Fermentation and Its Sensory Properties (수산물 첨가 김치의 이화학적 특성 변화 및 관능성)

  • Woo, Minji;Choi, Jung Ran;Kim, Mijeong;Jang, Mi-Soon;Cho, Eun Ju;Song, Yeong Ok
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.12
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    • pp.1771-1777
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    • 2012
  • This study was conducted to investigate the physicochemical characteristics of seafood added kimchi (SAK) during fermentation and its sensory properties. Korean cabbage kimchi (KCK) and four different SAKs were prepared and stored at $5^{\circ}C$ for eight weeks. The SAKs contained pre-treated octopus, squid, abalone, and webfoot octopus added at 12% (w/w) to the brined Korean cabbage. The fermentation patterns of SAKs were similar to those of KCK, indicating that the SAKs followed a typical fermentation process. Comparison of the physicochemical characteristics of SAKs with KCK revealed that the pH and acidity of SAKs was higher. The maximum concentrations of Lactobacillus spp. and Leuconostoc spp. for SAKs ranged from 8.31~8.85 and 7.60~8.14 log CFU/mL, respectively, which were higher than those for KCK. Therefore, the production of organic acids by microorganisms was greater in SAKs, which explained the higher acidities of the SAKs. Nitrogenous compounds hydrolyzed during fermentation, as well as reducing sugars and other nutritious compounds in SAKs might provide a good medium for lactic acid bacterial growth. Sensory evaluation was carried out using optimally ripened kimchi (pH $4.3{\pm}0.1$, acidity $0.7{\pm}0.1$), and the scores for sour taste, sour smell, and carbonated taste were significantly lower for SAKs than KCK. In the preference test, texture and overall acceptability were significantly higher for SAKs than KCK. Significant differences were not observed among SAKs upon subjective and preference evaluations. In conclusion, the fermentation patterns of SAKs were normal, regardless of seafood sources, and their sensory characteristics were comparable to or superior than those of KCK due to free amino acids, nitrogenous compounds produced during the fermentation, and reducing sugar present in the seafood.

Anti-inflammatory Activity of Antimicrobial Peptide Papiliocin 3 Derived from the Swallowtail Butterfly, Papilio xuthus (호랑나비 유래 항균 펩타이드 파필리오신 3의 항염증 활성)

  • Shin, Yong Pyo;Lee, Joon Ha;Kim, In-Woo;Seo, Minchul;Kim, Mi-Ae;Lee, Hwa Jeong;Baek, Minhee;Kim, Seong Hyun;Hwang, Jae Sam
    • Journal of Life Science
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    • v.30 no.10
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    • pp.886-895
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    • 2020
  • The development of novel peptide antibiotics with potent antimicrobial activity and anti-inflammatory activity is urgently needed. In a previous work, we performed an in-silico analysis of the Papilio xuthus transcriptome to identify putative antimicrobial peptides and identified several candidates. In this study, we investigated the antibacterial and anti-inflammatory activities of papiliocin 3, which was selected bioinformatically based on its physicochemical properties against bacteria and mouse macrophage Raw264.7 cells. Papiliocin 3 showed antibacterial activities against E. coli and S. aureus without inducing hemolysis and decreased the nitric oxide production of the lipopolysaccharide-induced Raw264.7 cells. Moreover, ELISA and Western blot analysis revealed that papiliocin 3 reduced the expression levels of pro-inflammatory enzymes, such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and prostaglandin E2 (PGE2). In addition, we examined whether papiliocin 3 could inhibit the expression of pro-inflammatory cytokines (interleukin-6 and interleukin-1β) in LPS-induced Raw264.7 cells. We found that papiliocin 3 markedly reduced the expression level of cytokines through the regulation of mitogen-activated protein kinases (MAPK) and nuclear factor kappa B (NF-κB) signaling. We also confirmed that papiliocin 3 binds to bacterial cell membranes via a specific interaction with lipopolysaccharides. Collectively, these findings suggest that papiliocin 3 could be a promising molecule for development as a novel peptide antibiotic.

Effect of the Fish Meat Hydrolysate on the Growth of Lactic Acid Bacteria (유산균(乳酸菌) 성장(成長)에 미치는 어육단백질분해물(魚肉蛋白質分解物)의 영향(影響))

  • Lee, Eun Bo;Kim, Jong Woo
    • Korean Journal of Agricultural Science
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    • v.11 no.1
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    • pp.120-132
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    • 1984
  • In order to clarify the effect of the fish meat hydrolysate on the growth of lactic acid bacteria(Str. lactis, Str. thermophibus, L. bulgaricus, L. acidophilus, and L. helveticus), the optimum conditions for hydrolyzing the fish meat were examined, and changes of the acid production, viable cell count of lactic acid bacteria and the charge of pH of the culture medium by addition of the fish meat hydrolysate were tested. The results were as follows: 1. When the hydrolysis of back muscle of mackerel was proceeded at $50^{\circ}C$ and at pH 8, for 48 hours adding 6% pancreatin of the protein content in the substrate, the best result was obtained. 2. The composition of the fish meat hydrolysate were 53.6% moisture, 32.4% protein, 1.0% fat, 10.7% carbohydrate, and 3.2% ash. 3. Above 0.1% of the fish meat hydrolysate in the culture medium, the acidity of the culture medium by Sir. lactis and Str. thermophilus were increased remarkably. The acidity of the culture medium by L. acidophilus and L. helveticus were increased in above 0.2% fish meat hydrolysate in the culture medium. but L. bulgaricus was not effected by the fish meat hydrolysate. 4. The pH of the culture medium during incubating Str. laclis and Sir. thermophilus failed obviously by adding the fish meat hydrolysate. But in the cases of L. bulgaricus, L. acidophilus, and L. helveticus, the pH were not changed clearly. 5. The viable cell count in all bacterial strains tested here were elevated by increasing the concentration of the fish meat hydrolysate.

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