• Title/Summary/Keyword: 분해균주

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Evaluation of Syringaresinol Content and MC3T3E1 Osteoblast Differentiation of Fermented Extracts of Eleutherococcus senticosus Using Lactobacillus ssp. with High 𝛽-Glucosidase Activity (𝛽-Glucosidase 활성이 높은 유산균을 이용한 한국가시오갈피 발효 추출물의 Syringaresinol의 함량 및 MC3T3E1조골세포 분화 평가)

  • Minji Kang;Minkyoung Kang;Sangnam Oh
    • Journal of Dairy Science and Biotechnology
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    • v.42 no.2
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    • pp.48-63
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    • 2024
  • This study investigated the 𝛽-glucosidase activity of lactic acid bacteria and specifically eleutheroside E and B from Acanthopanax senticosus after bioconverting them into syringaresinol (SYR). Out of 125 lactic acid bacteria strains isolated from kimchi and other sources, 46 exhibiting both extracellular and internal 𝛽-glucosidase activity were identified. Notably, strains LFFR 20-011 (Lactobacillus curvatus) and LFFR 20-043/LFR20-050 (Levilactobacillus brevis) enhanced SYR production by more than two-fold during Acanthopanax senticosus fermentation. Further investigation revealed that SYR significantly promoted osteoblast differentiation, as evidenced by the increased mRNA expression levels of early and mature osteoblast markers, including Runx2, type I collagen, and osteocalcin. These findings suggested that the enhanced presence of SYR through bioconversion by Acanthopanax senticosus may improve bone health. These results provide foundational data supporting the development of lactic-acid-bacteria-fermented Acanthopanax senticosus as a functional food aimed at promoting skeletal health in older adults.

Microbial Population Diversity of the Mud Flat in Suncheon Bay Based on 16S rDNA Sequences and Extracellular Enzyme Activities (남해안 갯벌 미생물의 세포외효소 활성 및 16S rDNA 분석에 의한 다양성 조사)

  • Kim, Yu-Jeong;Kim, Sung-Kyum;Kwon, Eun-Ju;Baik, Keun-Sik;Kim, Jung-Ho;Kim, Hoon
    • Applied Biological Chemistry
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    • v.50 no.4
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    • pp.268-275
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    • 2007
  • Diversity of the mud flat microbial population in Suncheon Bay was investigated by studying extracellular enzyme activities and 16S rDNA sequences. Four culturable bacterial strains with CMCase, xylanase and protease activities were isolated from the wetland and the mud flat. All the strains produced more xylanase activity than CMCase or protease activity, and the properties of the isolate enzymes from the wetland were similar to those from the mud flat. About 2,000 clones were obtained with the 16S rDNA amplified from the metagenomic DNA isolated from the mud samples. Based on the restriction pattern(s), seventeen clones were selected for base sequence analysis. Of the 17 clones, only 35% (6 clones) were found to be cultured strains and 65% (11 clones) to be uncultured strains. The similarities in the base sequences of the clones ranged from 91.0% to 99.9% with an average similarity of 97.3%. The clones could be divided into 7 groups, Proteobacteria (9 clones, 52.9%), Firmicutes (3 clones, 17.6%), Bacteroidetes (1 clone), Flavobacteria (1 clone), Verrucomicrobia (1 clone), Acidobacteria (1 clone), and Chloroflexi (1 clone). Most of the Proteobacteria clones were gamma Proteobacteria associated with oxidation-reduction of sulfur.

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 $30^{\circ}C$.

Biological Control of Sesame Soil-born Disease by Antifungal Microorganisms (참깨 토양전염성병(土壤傳染性病)의 생물학적방제(生物學的防除))

  • Shin, G.C.;Im, G.J.;Yu, S.H.;Park, J.S.
    • Korean journal of applied entomology
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    • v.26 no.4 s.73
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    • pp.229-237
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    • 1987
  • In order to study the biological control of soil-borne disease of sesame, antagonistic isolates of Trichoderma , Bacillus sand streptomyces to Fusarium oxysporum and Rhizoctonia solani were isolated from the rhizosphere soils of sesame plants and some other habitats. Out of the isolates of microorganisms collected a strain of Trichoderma viride was selected as a biological control agent for the study and its effect on the control of damping-off and the seedling growth of sesame was investigated. The results obtained are as follows: 26 percents of Bacillus spp. isolated from the rhizosphere soil of sesame plants showed antagonism to two pathogenic fungi. Important species were B. Subtilis and B. polymyxa. Streptomyces species isolated from the rhizosphere soils of sesame lysed the cell wall of hyphae and conidia of F. oxysporum and reduced conspicuously the formation of macroconidia and chlamydospores of the fungus. 84 percents of Trichoderma spp. isolated from the rhizosphere soil of sesame plants were antagonistic to F. oxysporum and 60 percents of the isolates were antagonistic to both F. oxysporum and R. solani. Trichoderma viride TV-192 selected from antagonistic isolates of Trichoderma spp. was highly antagonistic to F. oxysporum and soil treatment with the isolate reduced notably damping-off of sesame. T. viride TV-192 showed better growth in crushed rice straw, barley straw and sawdust media than F. oxysporum. Sawdust was selective for the growth of T. viride. Supplementation of wheat bran and mixtures of wheat bran and sawdust inoculated with T. viride TV-192 in the soil reduced remarkably damping-off of sesame by F. oxysporum but high density of the fungus TV-192 caused the inhibition of seed germination and seedling growth of sesame. Inhibitory effects of Trichoderma species on seed germination and seedling growth of sesame were different according to the isolates of the fungus. Normal sesame seedlings on the bed treated with the fungus showed better growth than not treated seedlings.

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The Synergy Effects of Mixed Treatment with Tannic Acid and Bacillus thuringiensis subsp. kurstaki KB100 against Spodoptera exigua (Lepidoptera: Noctuidae) (Tannic acid와 Bacillus thuringiensis subsp. kurstaki KB100균주의 혼합처리에 의한 파밤나방 살충활성의 상승효과)

  • Jin, Na-Young;Jung, Sun-Young;Park, Chan;Paek, Seung-Kyoung;Seo, Mi-Ja;Youn, Young-Nam;Yu, Yong-Man
    • Korean journal of applied entomology
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    • v.48 no.4
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    • pp.519-526
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    • 2009
  • Bacillus thuringiensis subsp. kurstaki KB100 isolated from the domestic soil have the most effective activity against the beet armyworm, Spodoptera exigua larva. The tannic acid as protease inhibitor might be increased the efficacy of sublethal concentrations of B. thuringiensis. The tannic acid was identified as a protease inhibitor that could increased the efficacy of sublethal concentrations of B. thuringiensis. Mixture of B. thuringiensis and tannic acid was investigated the mortality of S. exigua larva in the laboratory and field. When B. thuringiensis treated to 2nd larva of S. exigua, mortality was shown 54.4%. However, mixtures of B. thuringiensis with 4 and 40 mM tannic acid were increased mortalities to 2nd larva of S. exigua as 64.0 and 95.5%, respectively. Also, synergy effect of mixture of B. thuringiensis and 40 mM tannic acid was increased the mortality of S. exigua 3rd larva to 93.3%, even though 60.0% mortality with only B. thuringiensis treatment. On the other hand, the mortality of mixture with B. thuringiensis and 80 mM tannic acid was 53.3% lower than B. thuringiensis single treatment. In the welsh onion field, the accumulated mortalities of 3 times replicated with mixture of B. thuringiensis and 40 mM tannic acid were 83.9, 89.4 and 66.8% compare with 61.8, 80.4 and 47.3% as only B. thuringiensis treatment, respectively.

Characterization of Catechol l,2-Dioxygenase Purified from the Benzoate Degrading Bacterium, Pseudomonas sp. NFQ-l Isolated from Dead Coal Pit Areas (폐광지역에서 분리한 Benzoate 분해세균 Pseudomonas sp. NEQ-1에서 정제된 Catechol 1,2-Dioxygenase의 특성)

  • Joo Jung-Soo;Yoon Kyung-Ha
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.275-281
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    • 2004
  • Our previous research has demonstrated that the bacterium, Pseudomonas sp. NFQ-l capable of utilizing quin­oline (2,3-benzopyridine) as the sole source of carbon, nitrogen, and energy was isolated and characterized [Yoon et ai. (2003) Kor. J. Biotechnol. Bioeng. 18(3):174-179]. In this study, we have found that Pseudomonas sp. NFQ-l could degrade quinoline as well as benzoate, and extended this work to characterize the catechol 1,2­dioxygenase (C1,2O) purified from the bacterium cultured in benzoate media. Initially, C1,2O has been purified by ammonium sulfate precipitation, gel permeation chromatography, and Source 15Q. After Source 15Q, puri­fication fold was increased to approximately 14.21 unit/mg. Molecular weight of C1,2O was about 33 kDa. Physicochemical characteristics (e.g., substrate specificity, Km, Vmax, pH, temperature and effect of inhibitors) of purified C1,2O were examined. C1,2O demonstrated the activity for catechol, 4-methylcatechol and 3-meth­ylcatechol as a substrate, respectively. The Km and Vmax value of C1,2O for catechol was 38.54 ${\mu}M$ and $25.10\;{\mu}mol{\cdot}min^{-1}{\cdot}mg^{-1}.$ The optimal temperature of C1,2O was $30^{\circ}C$ and the optimal pH was approximately 8.5. Metal ions such as $Ag^+,\;Hg^+,\;Ca^{2+},\;and\;Cu^{2+}$ show the inhibitory effect on the activity of C1,2O. N-terminal amino sequence of C1,2O was analyzed as ^1TVKISQSASIQKFFEEA^{17}.$ In this work, we found that the amino acid sequence of NFQ-l showed the sequence homology of 82, 71, 59 and $53\%$ compared with C1,2O from Pseudomonas aeruginosa PA0l, Pseudomonas arvilla C-1., P. putida KT2440 and Pseudomonas sp. CA10, respectively.

Protoplast Formation, Regeneration and Reversion in Pleurotus ostreatus and P. sajor-caju (느타리버섯과 여름느타리버섯의 원형질체(原形質體) 나출(裸出)과 재생(再生))

  • Go, Seung-Joo;Shin, Gwan-Chull;Yoo, Young-Bok
    • The Korean Journal of Mycology
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    • v.13 no.3
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    • pp.169-177
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    • 1985
  • The studies were carried out to obtain the basic data for maximizing the protoplast yields from the mycelia of P. ostreatus and P. sajor-caju. Some factors affecting the regeneration of the protoplast of both species and the productivity of their reversion were also examined. The maximum yields of protoplasts were obtained from four days cultured mycelia of both species on cellophan membrane placed on the surface of PSA or MCM media in a petri dish. The optimal concentration of lytic enzyme Novozym 234 for protoplast releasing was 5 mg per ml of 0.5 M phosphate buffer solution with 0.6 M sucrose or 0.6 M $MgSO_4$ at pH 6.0. The greatest number of protoplasts was released 3 hours after incubation of the mycelia of P. ostreatus and after 4 hours for the P. sajor-caju in the lytic enzyme solution. Among the osmotic stabilizer solutions tested 0.6 M sucrose and 0.6 M KCl showed the best regeneration rates of the protoplasts of both species. When 0.75 % agar solution was over-layed on the regeneration media immediately after inoculation of the protoplast the regeneration rates were greatly enhanced. The ampicillin added to the agar solution prevented bacteria from infection. The reverted isolates produced the sporophores and basidial spores just like their parents without any mutations when they were cultivated in a broad mouth bottle with sawdust substrates.

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Studies on Microbial Inulase (Part I) -A Study on the Isolation of an Inulase Producing strain and the Optimum Cultural Conditions for the Enzyme Production- (미생물(微生物) Inulase에 관한 연구(硏究) 제1보(第一報) -Inulase생산균주(生産菌株)의 분리(分離)와 효소생산(酵素生産)을 위한 배양조건(培養條件)의 검토(檢討)-)

  • Kim, Ki-Choul
    • Applied Biological Chemistry
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    • v.18 no.1
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    • pp.42-51
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    • 1975
  • Penicillium sp I which produces a powerful hydrolysing enzyme was isolated from putrefid and dry Jerusalem artichoke medium. The strain was used to study on the optimum culture conditions for enzyme production. The results obtained are as follows: 1. Penicillium sp I was a vigorous strain to produce inulase. 2. The optimum culture conditions of the strain was examined in the Jerusalem artichoke extract medium and the synthetic medium. 3. Inulase productivity in the Jerusalem artichoke extract medium was higher than that of the synthetic medium. 4. The optimum culture period of the Jerusalem artichoke extract medium was four days, whereas that of the synthetic medium was five days. 5. The optimum temperature, pH and concentration in the Jerusalem artichoke extract medium were $30^{\circ}C$, 5.0 and 4.0% (W/V), respectively. Meanwhile, the optimum temperature, pH and concentration in the synthetic medium were $30{\sim}33^{\circ}C$, $5.0{\sim}6.0$, and $1.0{\sim}1.5%$ (W/V), respectively. 6. Corn steep liquor, peptone, $(NH_4)_2HPO_4,\;NH_4H_2PO_4,\;(NH_4)_2SO_4$, etc. were favorable as nitrogen sources. Of these, especially, Corn steep liquor and peptone as organic nitrogen sources caused an increase in inulase production in the synthetic medium. 7. All sugars except for inulin have no effect upon the inulase production. 8. KCl, $MgSO_4\;and\;FeSO_4$ were favourable mineral sources for inulase production.

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The Antimicrobial Characteristics of McSSP-31 Purified from the Hemocyte of the Hard-shelled Mussel, Mytilus coruscus (참담치(Mytilus coruscus) 혈구(hemocyte)에서 분리한 McSSP-31의 항균 특성 분석)

  • Oh, Ryunkyoung;Lee, Min Jeong;Kim, Young-Ok;Nam, Bo-Hye;Kong, Hee Jeong;Kim, Joo-Won;Park, Jung-Youn;Seo, Jung-Kil;Kim, Dong-Gyun
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1276-1289
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    • 2017
  • This study isolated and purified the antimicrobial peptide McSSP-31 from an acidified hemocyte extract of a Mytilus coruscus. The antimicrobial peptide was purified by using a $C_{18}$ reversed-phase high-performance liquid chromatography (HPLC). The peptide was determined to be 3330.549 Da by matrix assisted-laser desorption ionization time-of-flight mass spectrophotometer (MALDI-TOF/MS). The N-terminus of a 14 amino-acid sequence was identified as P-S-P-T-R-R-S-T-S-R-S-K-S-R by Edman degradation method. The acquired sequence showed a 93% similarity with the sperm-specific protein Phi-1, which is from M. californianus. The identified open-reading frame (ORF) of peptide was 306 bp encoding 101 amino acids, which was analyzed by rapid amplification of cDNA ends (RACE), cloning and sequencing analysis. We compared the full sequence with other known proteins that reveal the sperm-specific protein Phi-1 (93.5%) of M. californianus. Synthesized antimicrobial peptide (McSSP-31) showed antibacterial activity against gram-positive bacteria including B. subtilis, S. mutans, S. aureus and gram-negative bacteria including E. coli, K. pneumoniae, P. mirabilis, P. aeruginosa and fungi, C. albicans. Also, synthesized peptide showed strong antibacterial activity against antibiotic-resistant strains, including S. aureus. The cytotoxicity of the peptide was determined by using the HUVEC human cell line. The peptide did not exhibit any significant cytotoxic effects on the normal human cell line, and it had very low hemolytic activity with flounder hemoglobin. The results demonstrated that peptide purified from the hemocyte of a M. coruscus exhibits antibacterial activity against various bacteria and has the potential to be an alternative antibiotic agent.

Cold shock sensitive growth of Bacillus subtilis mutants deleted for genes involved in fatty acid synthesis (지방산 생합성 관련 유전자 결손 Bacillus subtilis 균주들의 저온충격 민감성 생장)

  • Kim, Do Hyung;Lee, Sang Soo
    • Korean Journal of Microbiology
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    • v.54 no.1
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    • pp.9-17
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    • 2018
  • To investigate the role of bkdR, sigL, yplP, and des genes which were known to be involved in fatty acid synthesis and sensitive at low temperature, deletion mutants of Bacillus subtilis CU1065 and JH642 were constructed. To determine the low temperature sensitivity of these genes, we compared the growth curves of cells at $37^{\circ}C$ and $15^{\circ}C$. At $37^{\circ}C$, wild type and deletion mutants showed almost similar growth but only bkdR deletion strain at $15^{\circ}C$ showed very slow growing compared with wild type. At $15^{\circ}C$ sigL and yplP deletions were somewhat slower or similar to those of wild type strain. Double and triple mutants for bkdR, sigL, yplP deletions were constructed and grown at $20^{\circ}C$ in LB agar to investigate cold sensitive growth. Double or triple deletions including bkdR deletion showed cold sensitive growing. In order to identify more clearly cold sensitive growth, the experiments were carried out under cold shock conditions in which the temperature was lowered from $37^{\circ}C$ to $15^{\circ}C$ at the point of 0.4 optical densities at 600 nm. In these cold shock experiments, only bkdR deletion showed significantly lower growing and additional des deletion increases cold sensitivity. The bkdR activates the bkd operon, which catabolized isoleucine, valine and leucine, amino acids and produce precursors for the synthesis of branched fatty acids. At cold shock growing of bkdR deletion strain, isoleucine recovered cold sensitivity of bkdR deletion but valine did not restore cold sensitivity. Isoleucine is used as a precursor for the synthesis of anteiso-branched fatty acids. On the other hand, valine is used as a precursor for the synthesis of iso-branched fatty acids. This indicates that anteiso-branched fatty acid plays an important role at the cold shock condition.