• Title/Summary/Keyword: Artificial enzyme

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Isolation, Identification, and Probiotic Properties of Lactobacillus reuteri HY701 from Human Feces

  • Kim, Jun-Tae;Jung, Hwang-Young;Lee, Na-Kyoung;Rhim, Seong-Lyul;Paik, Hyun-Dong
    • Food Science and Biotechnology
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    • v.15 no.5
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    • pp.677-682
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    • 2006
  • Strain HY701 was isolated from human feces for probiotic use by selecting highly resistant isolates to artificial gastric acid and bile acid. Strain HY701 was identified as Lactobacillus reuteri using 16S rDNA sequencing, and tentatively named L. reuteri HY701. The resistance of L. reuteri HY701 to artificial gastric acid (PH 2.5) was high with a survival rate of over 90%. L. reuteri HY701 also showed high tolerance to artificial bile acid after incubation in artificial gastric acid. Using the API ZYM test kit, the carcinogenic enzymes (${\beta}$-glucuronidase and (${\beta}$-glucosidase were not detected with L. reuteri HY70l, while the beneficial enzyme (${\beta}$-galactosidase was weakly detected. L. reuteri HY701 was sensitive to $100\;{\mu}g/mL$ nisin, $20\;{\mu}g/mL$ roxithromycin, $15\;{\mu}g/mL$ erythromycin, but resistant to $20\;{\mu}g/mL$ streptomycin, $10\;{\mu}g/mL$ tetracycline, $20\;{\mu}g/mL$ ciprofloxacin, $20\;{\mu}g/mL$ nystatin, $20\;{\mu}g/mL$ gentamycin, $10\;{\mu}g/mL$ doxycycline, $10\;{\mu}g/mL$ chloramphenicol, and $20\;{\mu}g/mL$ ampicillin. L. reuteri HY701 was shown to possess bactericidal activity as it inhibited the growth of Listeria monocytogenes ATCC 19111 and Escherichia coli JM109 completely within 24 hr of incubation. These results indicate that L. reuteri HY701 could be used as a probiotic strain.

Manipulation of Mini-Yeast Artificial Chromosome Containing Xylan Metabolism Related Genes and Mitotic Stability Analysis in Yeast (Xylan 대사유전자를가진미니효모인공염색체의가공및 Mitotic Stability 분석)

  • Da-In Kang;Yeon-Hee Kim
    • Microbiology and Biotechnology Letters
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    • v.50 no.3
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    • pp.436-440
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    • 2022
  • In this study, yeast artificial chromosome Insert (YAC) harboring genes which related xylan metabolism was constructed by using chromosome manipulation technique. For efficient chromosome manipulation, each splitting fragment (DNA module) required for splitting process was prepared and these DNA modules were transformed into Saccharomyces cerevisiae strain YKY164. By two-rounds chromosome splitting, yeast chromosome VII (1,124 kb) was split 887 kb-YAC, 45 kb-mini YAC and 198 kb-YAC and YKY183 strain containing 18 chromosomes was constructed. Splitting efficiency for chromosome manipulation was 50- 78% and expression level of foreign genes on 45 kb-mini YAC and enzyme activity were indistinguishable from that of the YKY164 strain. Furthermore, xylan-degraded products by recombinant enzymes were confirmed and mini-yeast artificial chromosome maintained stable mitotic stability without chromosome loss during 160 generations.

Production of Glutaminase (E.C. 3.2.1.5) from Zygosaccharomyces rouxii in Solid-State Fermentation and Modeling the Growth of Z. rouxii Therein

  • Iyer, Padma;Singhal, Rekha S.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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    • pp.737-748
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    • 2010
  • Glutaminase production in Zygosaccharomyces rouxii by solid-state fermentation (SSF) is detailed. Substrates screening showed best results with oatmeal (OM) and wheatbran (WB). Furthermore, a 1:1 combination of OM:WB gave 0.614 units/gds with artificial sea water as a moistening agent. Evaluation of additional carbon, nitrogen, amino acids, and minerals supplementation was done. A central composite design was employed to investigate the effects of four variables (viz., moisture content, glucose, corn steep liquor, and glutamine) on production. A 4-fold increase in enzyme production was obtained. Studies were undertaken to analyze the time-course model, the microbial growth, and nutrient utilization during SSF. A logistic equation ($R^2$=0.8973), describing the growth model of Z. rouxii, was obtained with maximum values of ${\mu}_m$ and $X_m$ at $0.326h^{-1}$ and 7.35% of dry matter weight loss, respectively. A goodfit model to describe utilization of total carbohydrate ($R^2$=0.9906) and nitrogen concentration ($R^2$=0.9869) with time was obtained. The model was used successfully to predict enzyme production ($R^2$=0.7950).

Expression and Purification of an ACE-Inhibitory Peptide Multimer from Synthetic DNA in Escherichia coli

  • OH, KWANG-SEOK;YONG-SUNG PARK;HA-CHIN SUNG
    • Journal of Microbiology and Biotechnology
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    • v.12 no.1
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    • pp.59-64
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    • 2002
  • An angiotensin I-converting enzyme (EC 3.4.15.1) (ACE), which can convert inactive angiotensin I into angiotensin II, a vasoconstrictor, is one of the key enzymes in controlling hypertension. It is suggested that the inhibition of ACE prevents hypertension, and many inhibitory peptides have already been reported. In the current study, oligonucleotides encoding ACE inhibitory peptides (IY, VKY) were chemically synthesized and designed to be multimerised due to isoschizomer sites (BamHI, BglII). The cloned gene named AP3 was multimerised up to 6 times in pBluescript and expressed in BL2l containing pGEX-KG. The fusion protein (GST-AP3) was easily purified with a high recovery by an affinity resin, yielding 38 mg of synthetic AP3 from a 1-1 culture. The digestion of AP3 by chymotrypsin exhibited an $IC_50$ value of $18.53{\mu}M$. In conclusion, the present experiment indicated that AP3 could be used as a dietary antihypertensive drug, since the potent ACE inhibitory activity of AP3 could be activated by chymotrypsin in human intestine.

Indirect enzyme linked immunosorbent assay for the diagnosis of brucellosis in cattle

  • Rahman, Siddiqur;Huque, Fazlul;Ahasan, Shamim;Song, Hee-Jong
    • Korean Journal of Veterinary Service
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    • v.33 no.2
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    • pp.113-119
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    • 2010
  • Brucellosis is a major zoonosis caused by Gram negative facultative intracellular bacterial organisms of the genus Brucella that are pathogenic for a wide variety of animals and human beings. Because of its economic impact on animal health and the risk to the human population,most countries have a brucellosis control program. Brucellosis is also an economically important andprevalent disease in Bangladesh. The accurate and prompt diagnosis is very important in controlling and eradicating of the disease in animals. The present study was undertaken to determine the seroprevalence of brucellosis in cattle in Mymensingh and Patuakhali district of Bangladesh. A total of 120 serum samples were collected from the two districts along with a questionnaire related to the epidemiology of the disease. The sampleswere screened by using slow agglutination test and conformed by indirect enzyme linked immunosorbent assay. The overall seroprevalence of brucellosis in cattle was 5% and it was observed that, a higher prevalence of Brucella was found in female than male, through natural breeding than artificial insemination (AI) and animal above 4 years old are highly susceptible than younger ones. Higher prevalence was found in aborted animals in comparison with non aborted animal. Finally, the study revealed that the female animal has more susceptible to brucellosis and healthy semen should be used for AI.

Application of Response Surface Methodology and Plackett Burman Design assisted with Support Vector Machine for the Optimization of Nitrilase Production by Bacillus subtilis AGAB-2

  • Ashish Bhatt;Darshankumar Prajapati;Akshaya Gupte
    • Microbiology and Biotechnology Letters
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    • v.51 no.1
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    • pp.69-82
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    • 2023
  • Nitrilases are a hydrolase group of enzymes that catalyzes nitrile compounds and produce industrially important organic acids. The current objective is to optimize nitrilase production using statistical methods assisted with artificial intelligence (AI) tool from novel nitrile degrading isolate. A nitrile hydrolyzing bacteria Bacillus subtilis AGAB-2 (GenBank Ascension number- MW857547) was isolated from industrial effluent waste through an enrichment culture technique. The culture conditions were optimized by creating an orthogonal design with 7 variables to investigate the effect of the significant factors on nitrilase activity. On the basis of obtained data, an AI-driven support vector machine was used for the fitted regression, which yielded new sets of predicted responses with zero mean error and reduced root mean square error. The results of the above global optimization were regarded as the theoretical optimal function conditions. Nitrilase activity of 9832 ± 15.3 U/ml was obtained under optimized conditions, which is a 5.3-fold increase in compared to unoptimized (1822 ± 18.42 U/ml). The statistical optimization method involving Plackett Burman Design and Response surface methodology in combination with an AI tool created a better response prediction model with a significant improvement in enzyme production.

Characteristics of Mycelial Growth and Enzyme Activities of Mattirolomyces terfezioides Collected from Robinia pseudoacacia Forest in Korea (국내 아까시나무 숲에서 수집한 감자덩이버섯(Mattirolomyces terfezioides)의 균사 생장 특성과 효소 활성)

  • Jeon, Sung-Min;Wang, Eun-Jin;Ka, Kang-Hyeon
    • The Korean Journal of Mycology
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    • v.43 no.3
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    • pp.165-173
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    • 2015
  • Mattirolomyces terfezioides is a type of sweet truffles that predominantly form ectomycorrhizae with Robinia pseudoacacia. It is also worthy of artificial cultivation. This is the first report on characteristics of mycelial growth and enzyme activities of M. terfezioides collected from R. pseudoacacia forest in Korea. M. terfezioides showed the highest mycelial growth when cultured on potato dextrose agar (PDA) at $30^{\circ}C$ or in modified Melin-Norkran's liquid medium (pH 8.0). The biomass of M. terfezioides was higher in liquid medium containing nitrate-nitrogen than ammonium-nitrogen by 1.8 fold. The mycelia of M. terfezioides showed both carboxymethylcellulase and laccase activities on solid media for enzyme screening.

Differences of Soil Enzyme Activity after Incorporation with Chinese Milk Vetch Litter Cut at Different Growth Stages

  • Lee, Ji-Hyun;Shim, Sang-In
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.52 no.3
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    • pp.341-347
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    • 2007
  • Chinese milk vetch (CMV) is a winter legume that is commonly used as cover crop in Korea. Kill date of cover crop for addition into soil affects N content in cover crop and N availability in soil. This study was conducted to evaluate the effect of CMV as green manure cover crop according to kill dates before growing corn without artificial fertilizer. Top of CMV cut three times on 13 April, 27 April, and 11 May were added into soil at a rate of 600 kg per 10a. Sugar content in CMV litter was persistently decreased from mid-April to late-May. The decrease of sugar content might be due to the transformation into starch and/or other storage or structural constituents. The decreased amount of sugars was greater than 12% and the increased amount of starch was less than 0.2%. Concentration of $NH_4^+$ in soil treated by CMV litter cut on May 11 was slightly higher than that in the treatment with early-cut (April 13) CMV, the concentration at 28 and 49 DAT (days after treatment) was higher in the treatment with late-cut CMV litter. Regardless of cut (kill) date of CMV, the phosphatase activity in the treatment of CMV litter was higher compared to the untreated control. Soil dehydrogenase activity was increased steadily by addition of CMV litter implying total microbial activities in the soil were increased. Our results demonstrate that the status of cover crop species at kill date is an important factor influencing soil enzyme activities derived from microorganisms. Therefore, the optimal kill date of cover crop should be examined to improve the efficiency of cover crop as green manure crop regarding the practical sequence in cropping system.

The Activity and Structure of Bacterial Community within Artificial Vegetation Island (AVI) (인공 수초재배섬에서 세균의 활성과 세균 군집 구조)

  • Jeon, Nam-Hui;Park, Hae-Kyung;Byeon, Myeong-Seop;Choi, Myung-Jae
    • Journal of Korean Society on Water Environment
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    • v.23 no.5
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    • pp.676-682
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    • 2007
  • The bacterial number, extracellular enzyme activities and structure of bacterial community which are major constituent of aquatic ecosystem within the artificial vegetation island (AVI) were compared to those of the nearby pelagic lake waters in order to evaluate the possibility of the AVI as a eco-technological measure for water quality improvement and restoration of littoral zone in man-made reservoirs. There was not a significant difference in the total number of bacteria, but the number of active (viable) bacteria within the AVI was about 0.7 to 4.1 times higher than nearby pelagic lake water. The ratio of the number of active bacteria versus the total number of bacteria was also higher in the AVI than nearby pelagic lake water. The activities of ${\beta}$-glucosidase and phosphatase were 1.0 to 13.1 and 0.8 to 7.3 times higher respectively in the AVI than nearby pelagic lake water, showing that microorganisms were more active within the AVI. The bacterial communities of the two waters, examined by FISH method, did not indicate a clear difference in the springtime when the growth of macrophytes was immature, but during summer and fall it showed a clear difference indicating the formation of distinct bacterial community within the AVI compared to nearby lake water. From the results of this study, we conclude that AVI can contribute to make up the littoral ecosystem which show rapid cycling of matters through active detritus food chain in the dam reservoirs which have unstable aquatic ecosystem due to short hydraulic residence time and to strengthen the self-purification capacity of the lake.

Activity of Mixed Function Oxidase in a few Insect Species in Relation to Their Food Source (먹이종류에 따른 몇가지 곤충의 MFO활성 비교)

  • 이정호;부경생
    • Korean journal of applied entomology
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    • v.32 no.3
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    • pp.291-299
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    • 1993
  • Midgut tissues from 4 insect specIes were exammed for the activity of cytochrome P-450 monooxygenases, a major enzyme involved in chemical detoxification. When Helicoverpa assulta larvae were reared on an artificial d;et, the specific activity of the midgut cytochrome P-450 monooxygenases (MFO) was :3 times higher than that of the fat body, The specific activity of the midgut cytochrome P-450 monooxygenases was higher in H. assul/a larvae when reared on Nicotiana tabacum leaves than when on CapsIcum annuum fruits or an artificial diet. In the case of Hyphantria cunea larvae, Tilia megaphyllo leaves were the best in inducing midgut cytochrome P-450 monooxygenases activity. When larvae of H. assulta, Spodoptera exigua, H. cunea and Spodoptera litura were reared on their own artificial diet, the highest activity was seen in S. exigua larvae which is a polyphagous and insecticide-resistant strain.

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