• Title/Summary/Keyword: Microbial Biotechnology

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Bioinformatics based Identification and Characterization of Epoxide Hydrolase of Gordonia westfalica for the Production of Chiral Epoxides (Bioinformatics를 활용한 토양미생물인 Gordonia westfalica Epoxide Hydrolase 생촉매 개발 및 Chiral Epoxides 제조 특성 분석)

  • Lee Soo Jung;Lee Eun Jung;Kim Hee Sook;Lee Eun Yeol
    • KSBB Journal
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    • v.20 no.4
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    • pp.311-316
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    • 2005
  • Epoxide hydrolases (EHs) are versatile biocatalysts for the preparation of chiral epoxides by enantioselective hydrolysis from racemic epoxides. Various microorganisms were identified to possess a EH activity by multiple sequence alignment and analysis of conserved domain sequence from genomic and megaplasmid sequence data. We successfully isolated Gordonia westfalica possessing EH activity from various microbial strains from culture type collections. G. westfalica exhibited (R)-styrene oxide preferred enantioselective hydrolysis activity. Chiral (S)-styrene oxide with high optical purity $(>\;99\%)\;ee)$ and yield of $36.5\%$ was obtained from its racemate using whole-cell of G. westfalica.

Induction of Arabidopsis thaliana Chitinase by Ethylene and Elicitor Treatment (에틸렌 및 Elicitor처리에 의한 아기장대풀의 키틴 가수분해 효소 유도)

  • Kyung Hee PAEK;Seok Yoon KWON;Hye Sun CHO;Jin Sam YOU
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.6
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    • pp.357-362
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    • 1994
  • Chitinases and $\beta$-1,3-glucanases are believed to be important in defending plane against pathogens. Here, we investigated the expression of chitinase(s) in Arabidopsis thaliana cell suspension culture system in response to ethephon (2-chloroethyl phosphonic acid) which produces ethylene or a microbial elicitor, a bacterial pectin-degrading enzyme, $\beta$-1, 4-endopolygalactronic acid Iyase (PGA Iyase), treatment. Chitinase activity was measured either by radio chemical assay using $^3$H-labeled regenerated chitin as substrate or western blot analysis using antibody raised against tobacro chitinase(S). With 1 mg/mL of ethephon or 100 m units/mL of elicitor treatment, maximum levels of activity were reached after 48h. We also investigated distribution of chitinase activity in seedlings, leaves, and root of A. thaliana and found that root have the highest chitinase activity.

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Acrolein with an α,β-unsaturated Carbonyl Group Inhibits LPS-induced Homodimerization of Toll-like Receptor 4

  • Lee, Jeon-Soo;Lee, Joo Young;Lee, Mi Young;Hwang, Daniel H.;Youn, Hyung Sun
    • Molecules and Cells
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    • v.25 no.2
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    • pp.253-257
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    • 2008
  • Acrolein is a highly electrophilic ${\alpha},{\beta}$-unsaturated aldehyde present in a number of environmental sources, especially cigarette smoke. It reacts strongly with the thiol groups of cysteine residues by Michael addition and has been reported to inhibit nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) activation by lipopolysaccharide (LPS). The mechanism by which it inhibits $NF-{\kappa}B$ is not clear. Toll-like receptors (TLRs) play a key role in sensing microbial components and inducing innate immune responses, and LPS-induced dimerization of TLR4 is required for activation of downstream signaling pathways. Thus, dimerization of TLR4 may be one of the first events involved in activating TLR4-mediated signaling pathways. Stimulation of TLR4 by LPS activates both myeloid differential factor 88 (MyD88)- and TIR domain-containing adapter inducing $IFN{\beta}$ (TRIF)-dependent signaling pathways leading to activation of $NF-{\kappa}B$ and IFN-regulatory factor 3 (IRF3). Acrolein inhibited $NF-{\kappa}B$ and IRF3 activation by LPS, but it did not inhibit $NF-{\kappa}B$ or IRF3 activation by MyD88, inhibitor ${\kappa}B$ kinase $(IKK){\beta}$, TRIF, or TNF-receptor-associated factor family member-associated $NF-{\kappa}B$ activator (TANK)-binding kinase 1 (TBK1). Acrolein inhibited LPS-induced dimerization of TLR4, which resulted in the down-regulation of $NF-{\kappa}B$ and IRF3 activation. These results suggest that activation of TLRs and subsequent immune/inflammatory responses induced by endogenous molecules or chronic infection can be modulated by certain chemicals with a structural motif that enables Michael addition.

Quality Characterization of Red Bean Gochujang prepared with Different Ratios of Meju (고추장용 메주 배합비를 달리하여 제조한 팥고추장의 품질 특성)

  • Kwon, Nu Ri;Yoon, Hyang-Sik;Kim, Ik Jei;Hong, Seong Taek;Kim, So-Young;Gil, Na Young;Han, Nam Soo;Eom, Hyun-Ju
    • The Korean Journal of Food And Nutrition
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    • v.31 no.5
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    • pp.751-759
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    • 2018
  • This study was conducted to investigate quality characteristics of red bean gochujang prepared with different ratios of meju for eight weeks. We determined quality characteristics such as pH, moisture content, total microbial flora counts, amino-type nitrogen content, total polypenol content, and antioxidant activity. The pH decreased by fermentation period, whereas total acidity was 0.36~0.39% in the early stage of fermentation, 0.63~1.16% at four weeks of fermentation, and decreased to 0.43~0.65% after the eighth week of fermentation. Moisture content of the control (no red bean, no rice) decreased 62.13% to 50.93%, but in case of the treatment, it slightly decreased. Total cell counts revealed similar tendency at the beginning of fermentation, and at eight weeks of fermentation. In case of lactic acid bacteria, all samples except RB-1.5 (gochujang added meju made of mixing ratio of rice:soybean:red bean (1:2:1.5) slightly increased. Amino nitrogen and total polyphenol of all samples increased in the fermentation period, especially RB-1.5 sample had the highest levels than other samples. By increasing the amount of red bean, DPPH radical scavenging activities increased. In taste sensing analysis, all treat samples except RB-1.5 revealed lower sourness than the control. Also, in case of bitterness, all samples revealed low content than the control. So, by adding red bean, it can enhance taste and function of gochujang.

Rumen Microbes, Enzymes and Feed Digestion-A Review

  • Wang, Y.;McAllister, T.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.11
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    • pp.1659-1676
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    • 2002
  • Ruminant animals develop a diverse and sophisticated microbial ecosystem for digesting fibrous feedstuffs. Plant cell walls are complex and their structures are not fully understood, but it is generally believed that the chemical properties of some plant cell wall compounds and the cross-linked three-dimensional matrix of polysaccharides, lignin and phenolic compounds limit digestion of cell wall polysaccharides by ruminal microbes. Three adaptive strategies have been identified in the ruminal ecosystem for degrading plant cell walls: production of the full slate of enzymes required to cleave the numerous bonds within cell walls; attachment and colonization of feed particles; and synergetic interactions among ruminal species. Nonetheless, digestion of fibrous feeds remains incomplete, and numerous research attempts have been made to increase this extent of digestion. Exogenous fibrolytic enzymes (EFE) have been used successfully in monogastric animal production for some time. The possibility of adapting EFE as feed additives for ruminants is under intensive study. To date, animal responses to EFE supplements have varied greatly due to differences in enzyme source, application method, and types of diets and livestock. Currently available information suggests delivery of EFE by applying them to feed offers the best chance to increase ruminal digestion. The general tendency of EFE to increase rate, but not extent, of fibre digestion indicates that the products currently on the market for ruminants may not be introducing novel enzyme activities into the rumen. Recent research suggests that cleavage of esterified linkages (e.g., acetylesterase, ferulic acid esterase) within the plant cell wall matrix may be the key to increasing the extent of cell wall digestion in the rumen. Thus, a crucial ingredient in an effective enzyme additive for ruminants may be an as yet undetermined esterase that may not be included, quantified or listed in the majority of available enzyme preparations. Identifying these pivotal enzyme(s) and using biotechnology to enhance their production is necessary for long term improvements in feed digestion using EFE. Pretreating fibrous feeds with alkali in addition to EFE also shows promise for improving the efficacy of enzyme supplements.

Antimicrobial Characteristic of Prunus mune extract (매실추출물의 항균특성)

  • Ha Myung-Hee;Park Woo-Po;Lee Seung-Cheol;Choi Sung-Gil;Cho Sung-Hwan
    • Food Science and Preservation
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    • v.13 no.2
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    • pp.198-203
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    • 2006
  • Prunus mume extract showed antimicrobial efface remarkably against the wide spectrum of putrefactive and food spoilage microorganisms above 250 ppm of concentration. Their thermal and pH stabilities were effective under the range of temperature $(40^{\circ}C{\sim}120^{\circ}C)$ and $pH(3{\sim}11)$. Prunus mume extract seemed to be a natural antimicrobial ideally with the view of their effectiveness and thermal & pH stabilities. In addition, their action modes suggested that their hydrophillic components would perturb the fucntions of microbial cell membranes synergistically.

Biocatalytic Production of Chiral Epoxides (생촉매를 이용한 광학활성 에폭사이드 생산)

  • 이은열;최원재;윤성준;김희숙;최차용
    • KSBB Journal
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    • v.14 no.3
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    • pp.291-296
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    • 1999
  • Chiral epoxides are key intermediates for the production of chiral pharmaceuticals, agrochemicals, and functional food additives. Chiral epoxides can be produced by either chemical or biological method. In biocatalytic production routes, chiral epoxides can be produced via epoxidations of prochiral alkenes by monooxygenase or peroxidase. Kinetic resolution of racemic epoxides using whole cells of bacteria or fungi might be commercially useful, since it is possible to obtain chiral epoxides with high optical purities from relatively cheap and readily avaiable racemic epoxides. Some bioprocesses already are commercially developed: the biocatalytic production of chiral epichlorohydrin via microbial stereospecific dehalogenation, and lipase-catalyzed enantioselective hydrolysis in a hollow fiber membrane bioreactor for the production of chiral methyl trans-3-(4-methoxyphenyl)glycidate. the intermediate for calcium antagonist diltiazem. The importance of biocatalytic production of chiral epoxides with several examples from literature are presented.

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Clearance of Cervical Human Papillomavirus Infection by Topical Application of Curcumin and Curcumin Containing Polyherbal Cream: A Phase II Randomized Controlled Study

  • Basu, Partha;Dutta, Sankhadeep;Begum, Rakiba;Mittal, Srabani;Dutta, Paromita Das;Bharti, Alok Chandra;Panda, Chinmay Kumar;Biswas, Jaydip;Dey, Bindu;Talwar, Gursaran Prashad;Das, Bhudev Chandra
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.5753-5759
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    • 2013
  • Curcumin and curcumin containing polyherbal preparations have demonstrated anti-microbial and antiviral properties in pre-clinical studies. Till date no therapeutic intervention has been proved to be effective and safe in clearing established cervical human papillomavirus (HPV) infection. The present study evaluated the efficacy of Basant polyherbal vaginal cream (containing extracts of curcumin, reetha, amla and aloe vera) and of curcumin vaginal capsules to eliminate HPV infection from cervix. Women were screened by Pap smear and HPV DNA test by PCR. HPV positive women without high grade cervical neoplasias (N=287) were randomized to four intervention arms to be treated with vaginal Basant cream, vaginal placebo cream, curcumin vaginal capsules and placebo vaginal capsules respectively. All subjects were instructed to use one application of the assigned formulation daily for 30 consecutive days except during menstruation and recalled within seven days of the last application for repeat HPV test, cytology and colposcopy. HPV clearance rate in Basant arm (87.7%) was significantly higher than the combined placebo arms (73.3%). Curcumin caused higher rate of clearance (81.3%) than placebo though the difference was not statistically significant. Vaginal irritation and itching, mostly mild to moderate, was significantly higher after Basant application. No serious adverse events were noted.

Bypass Fat Production Using Acid Oil, Its Effect on In Vitro Rumen Fermentation and Effect of Its Feeding on In Sacco DM Disappearance in Sheep

  • Garg, M.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.10 no.6
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    • pp.571-574
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    • 1997
  • Attempts were made in the laboratory to produce bypass fat using acid oil by precipitation and fusion methods. The degree of saponification by both of these methods was above 80 percent. Where heating facilities are not available, precipitation method could be used, otherwise, fusion method of bypass fat production is found to be more convenient, especially for commercial scale operations as handling of large volume of solutions is eliminated. Bypass fat thus produced was tested in vitro for rumen fermentation. Incorporation of acid oil in the incubation medium reduced TVFA conc. from 127.06 to 124.09 mM/l SRL and increased ammonia-N levels from 210.50 to 223 mg/l SRL indicating that the microbial activity was affected on incorporation of acid oil in the incubation medium. However, incorporation of bypass fat in the incubation medium did not significantly affect TVFA conc. as well as ammonia-N levels. In another experiment, nine rumen fistulated sheep in three groups of three each were fed bypass fat at two different levels. Dry matter disappearance in 24 h from the nylon bags suspended in the rumen of animals under different groups was found to be $47.74{\pm}1.10$, $47.55{\pm}0.21$ and $50.74{\pm}1.11$ in group I (control), group II (fed bypass fat 50 g/day) and group III (fed bypass fat 100 g/day), respectively. These studies indicated that it is possible to produce bypass fat from acid oils, a by-product of oil refining process, and its feeding did not affect rumen fermentation.

Characteristic of whole cell benzoylformate decarboxylase from Pseudomonas putida (Pseudomonas putida에서 생산된 전세포 benzoylformate decarboxylase의 활성특성 및 고정화 캡슐 제조)

  • 정재용;하태욱;홍진혁;오창엽;박중곤
    • KSBB Journal
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    • v.14 no.3
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    • pp.264-272
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    • 1999
  • Benzoylformate was converted to benzaldehyde by whole cell enzyme from Pseudomonas putida KCTC 1751. We investigated the effect of the composition of the growth medium on th accumulation of benzoylformate decarboxylase in the microbial cell. We prepared a calcium alginate capsule containing Pseudomonas putida cells to develop a reusable whole cell enzyme. Pseudomonas putida cells were inoculated in the capsule and cultured in M1 medium for 1 day followed by cultivation in M3 medium for 3 days. The dry cell density reached 77.75 g/L on the basis of the inner volume of the capsule. The specific activity of encapsulated whole cell benzoylformate decarboxylase was half as high as that of free whole cell enzyme. The activity of encapsulated whole cell benzoylformate decarboxylase was half as high as that of free whole cell enzyme. The activity of encapsulated whole cell benzoylformate decarboxylase decreased 20 % after use for 20 batches and 40% after use for 30 batches. The dry cell density reduced about 10 % after 30 trials.

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