• Title/Summary/Keyword: hydrolytic activity

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Isolation and Identification of Low Molecular Weight Compounds Produced by Bacillus subtilis HJ927 and Their Biocontrol Effect on the Late Blight of Pepper (Capsicum annuum L.) (Bacillus subtilis HJ927에 의해 생성된 화합물의 분리, 동정 및 고추(Capsicum annum L.) 역병방제 효과)

  • Lee, Hyun-Jin;Park, Keun-Hyung;Shim, Jae-Han;Park, Ro-Dong;Kim, Yong-Woong;Hwang-Bo, Hoon;Cho, Jeung-Yong;Kim, Young-Cheol;Kim, Kil-Yong
    • Korean Journal of Soil Science and Fertilizer
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    • v.38 no.1
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    • pp.25-31
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    • 2005
  • A soil bacterium, Bacillus subtilis HJ927, exhibiting strong antagonistic property against pathogenic fungi was isolated from pepper fields infested with Phytophthora capsici. Pepper plants inoculated with P. capsici revealed severe root mortality while plants co-inoculated with B. subtilis HJ927 and P. capsici showed drastically reduced root mortality. Low molecular weight substances released by B. subtilis HJ927 mediated the plant protective effect. The anti-fungal compounds released by B. subtilis HJ927 were identified as 3-methylbutyric acid, 2-methylbutyric acid, and methyl 2-hydroxy, 3-phenylpropanoate by high-performance liquid chromatography and gas chromatography-mass spectrometry. In addition to these compounds, B. subtilis HJ927 also produced ${\beta}$-1,3-glucanase, a hydrolytic enzyme implicated in antifungal activity.

Processing and Biological Activity of Gelatin Hydrolysate from Branchiostegus japonicus Scales (옥돔(Branchiostegus japonicus) 비늘 유래 젤라틴의 가수분해 및 가수분해물의 기능성)

  • Ahn, Yong-Seok;Lee, Won-Woo;Lee, Seung-Hong;Ahn, Gin-Nae;Ko, Chang-Ik;Oh, Chang-Kyung;Oh, Myung-Cheol;Kim, Dong-Woo;Jeon, You-Jin;Kim, Soo-Hyun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.42 no.5
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    • pp.417-425
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    • 2009
  • The potential utility of fish scales to the functional food industry has been investigated due to its antioxidant and antihypertensive characteristics. In this study, we report on the reactive oxygen species (ROS) scavenging and angiotensin I converting enzyme (ACE) inhibitory activities of gelatin hydrolysates processed from Branchiostegus japonicus scales, which are also high in protein content (about 46.1%). We prepared the enzymatic gelatin hydrolysates with four proteases (${\alpha}$-chymotrypsin, Alcalase, Neutrase and trypsin) from B. japonicus scale gelatin, which was prepared according to different reaction times, substrate/enzyme ratios and substrate concentrations. The enzymatic hydrolytic degrees of the gelatin increased time-dependently up to 6 hrs, while the Alcalase gelatin hydrolysates showed the highest hydrolysis degrees compared to the others. Furthermore, gelatin hydrolysates of Neutrase and ${\alpha}$-chymotrypsin showed the highest DPPH radical and $H_2O_2$ scavenging activities ($IC_{50}$ value; 9.18 mg/mL and 9.74 mg/mL), respectively. However, the activities were not significant (P<0.05). We also observed that the four gelatin hydrolysates significantly increased ACE inhibitory activities from approximately 20% to 60% (P<0.05), Among them, the Alcalase gelatin hydrolysates showed the higher ACE inhibitory activity ($IC_{50}$ value; 0.73 mg/mL) compared to the others. These results suggest that the enzymatic gelatin hydrolysates prepared from B. japonicus scales may possess a potentially useful function as an ACE inhibitory agent. As such, the utility of B. japonicus scales should be given due consideration for application in the functional food industry.

Characterization of Single Stranded DNA-Dependent ATPase Activities of Deinococcus radiodurans RecA Protein (Deinococcus radiodurans RecA 단백질의 외가닥 DNA-의존성 ATPase 활성 분석)

  • Kim, Jong-Il
    • Korean Journal of Microbiology
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    • v.43 no.4
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    • pp.250-255
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    • 2007
  • The RecA protein of Deinococcus radiodurans is essential for the extreme radiation resistance of this organism. The central steps involved in recombinational DNA repair require DNA-dependent ATP hydrolysis by recA protein. Key feature of RecA protein-mediated activities is the interactions with ssDNA and dsDNA. The ssDNA is the site where RecA protein filament formation nucleates and where initiation of DNA strand exchange takes place. The effect of sequence heterogeneity of ssDNA was examined in this experiment. The rate of homopolymeric synthetic ssDNA-dependent ATP hydrolysis was constant or nearly so over a broader range of pHs. For poly(dT)-dependent ATP or dATP hydrolysis, rates were generally faster, with a broader optimum between pH 7.0 and 8.0. Activities of RecA protein were affected by the ionic environment. The ATPase activity was shown to have different sensitivity to anionic species. The presence of glutamate seemed to slimulate the hydrolytic activity. Dr RecA protein was shown to require $Mg^{2+}$ ion greater than 2 mM for binding to etheno ssDNA and the binding stoichiometry of 3 nucleotide for RecA protein monomer.

Molecular Cloning and Characterization of a Novel Exo-β-1,3-Galactanase from Penicillium oxalicum sp. 68

  • Zhou, Tong;Hu, Yanbo;Yan, Xuecui;Cui, Jing;Wang, Yibing;Luo, Feng;Yuan, Ye;Yu, Zhenxiang;Zhou, Yifa
    • Journal of Microbiology and Biotechnology
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    • v.32 no.8
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    • pp.1064-1071
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    • 2022
  • Arabinogalactans have diverse biological properties and can be used as pharmaceutical agents. Most arabinogalactans are composed of β-(1→3)-galactan, so it is particularly important to identify β-1,3-galactanases that can selectively degrade them. In this study, a novel exo-β-1,3-galactanase, named PoGal3, was screened from Penicillium oxalicum sp. 68, and hetero-expressed in P. pastoris GS115 as a soluble protein. PoGal3 belongs to glycoside hydrolase family 43 (GH43) and has a 1,356-bp gene length that encodes 451 amino acids residues. To study the enzymatic properties and substrate selectivity of PoGal3, β-1,3-galactan (AG-P-I) from larch wood arabinogalactan (LWAG) was prepared and characterized by HPLC and NMR. Using AG-P-I as substrate, purified PoGal3 exhibited an optimal pH of 5.0 and temperature of 40℃. We also discovered that Zn2+ had the strongest promoting effect on enzyme activity, increasing it by 28.6%. Substrate specificity suggests that PoGal3 functions as an exo-β-1,3-galactanase, with its greatest catalytic activity observed on AG-P-I. Hydrolytic products of AG-P-I are mainly composed of galactose and β-1,6-galactobiose. In addition, PoGal3 can catalyze hydrolysis of LWAG to produce galacto-oligomers. PoGal3 is the first enzyme identified as an exo-β-1,3-galactanase that can be used in building glycan blocks of crucial glycoconjugates to assess their biological functions.

Structure activity relationship on the herbicidal activity by the N-phenyl substituents of 2-(4-(6-chloro-2-benzoxazolyloxy)phenoxy)-N-Phenylpropionamide derivatives in down land (수답에서 2-(4-(6-chloro-2-benz-oxazolyloxy)phenoxy)-N-phenylpropionamide 유도체 중 N-phenyl 치환체들의 제초활성)

  • Sung, Nack-Do;Lee, Sang-Ho;Ko, Young-Kwan;Lee, Kyung-Mo;Kim, Dae-Whang;Kim, Tae-Joon
    • The Korean Journal of Pesticide Science
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    • v.4 no.2
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    • pp.21-28
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    • 2000
  • A new fourty six 2-(4-(6-chloro-2-benzoxazolyloxy)phenoxy)-N-phenylpro- pionamide derivatives were synthesized and the herbicidal activities against rice plant and barnyard grass with pre-emergence in down land were measured. The structure activity relationships (SAR) between the activities and physicochemical parameters of the substituted(X) N-phenyl group in substrates were analyzed and discussed by Free- Wilson and Hansch method from the basis on the former study (Sung. et. al., 1999). The conditions of selective herbicide activity both the barnyard grass and rice plant are shown that the optimal hydrophobicity, $({\pi})_{opt.}=1.34$ and electron donating with field effect (F<0) of meta and ortho, para-substituted mono or disubstituent on the N-phenyl ring were found to contribute significantly. The herbicidal activities against barnyard grass are roughly the same as the results in up land whereas damage to rice plant in down land more increase than that of up land. Degradation products in water are 2-(4-(6-chloro-2-benzoxazolyloxy)phenoxy)propionic acid ((A)) (obs. pKa=4.35 & obs. logP=4.77) and 6-chloro-2-benzoxazolone (B) (obs. pKa=8.40 & obs. logP=2.90). These results were supposing that the hydrolysis product of substrates, (A) is comparatively absorbed in rice plant but not in barnyard grass. And it is assumed from the SAR equations that the 2,6-dimethyl-4-methoxymethyl group substituent ($pI_{50}=5.41$, 3g/ha) is selected as the most highest herbicidal activity against barnyard grass in green house.

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A novel cold-active lipase from Psychrobacter sp. ArcL13: gene identification, expression in E. coli, refolding, and characterization (새로운 Psychrobacter sp. ArcL13 유래 저온활성 지질분해효소 : 유전자 분리동정, 대장균에서의 발현, refolding 및 특성 연구)

  • Koo, Bon-Hun;Moon, Byung-Hern;Shin, Jong-Suh;Yim, Joung-Han
    • Korean Journal of Microbiology
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    • v.52 no.2
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    • pp.192-201
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    • 2016
  • Recently, Psychrobacter sp. ArcL13 strain showing the extracellular lipase activity was isolated from the Chuckchi Sea of the Arctic Ocean. However, due to the low expression levels of the enzyme in the natural strain, the production of recombinant lipase is crucial for various applications. Identification of the gene for the enzyme is prerequisite for the production of the recombinant protein. Therefore, in the present study, a novel lipase gene (ArcL13-Lip) was isolated from Psychrobacter sp. ArcL13 strain by gene prospecting using PCR, and its complete nucleotide sequence was determined. Sequence analysis showed that ArcL13-Lip has high amino acid sequence similarity to lipases from bacteria of some Psychrobacter genus (84-90%) despite low nucleotide sequence similarity. The lipase gene was cloned into the bacterial expression plasmid and expressed in E. coli. SDS-PAGE analysis of the cells showed that ArcL13-Lip was expressed as inclusion bodies with a molecular mass of about 35 kDa. Refolding was achieved by diluting the unfolded protein into refolding buffers containing various additives, and the highest refolding efficiency was seen in the glucose-containing buffer. Refolded ArcL13-Lip showed high hydrolytic activity toward p-nitrophenyl caprylate and p-nitrophenyl decanoate among different p-nitrophenyl esters. Recombinant ArcL13-Lip displayed maximal activity at $40^{\circ}C$ and pH 8.0 with p-nitrophenyl caprylate as a substrate. Activity assays performed at various temperatures showed that ArcL13-Lip is a cold-active lipase with about 40% and 73% of enzymatic activity at $10^{\circ}C$ and $20^{\circ}C$, respectively, compared to its maximal activity at $40^{\circ}C$.

Behaviour of the Soil Residues of the Acaricide-Insecticide, [$^{14}C$]Acrinathrin;I. Behaviour during Crop(Maize) Cultivation (살비살충제 [$^{14}C$Acrinathrin 토양 잔류물의 행적 규명;I. 농작물(옥수수) 재배시의 행적)

  • Lee, Jae-Koo;Kyung, Kee-Sung;Kwon, Jeong-Wook;Ahn, Ki-Chang;Jung, In-Sang
    • Korean Journal of Environmental Agriculture
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    • v.14 no.2
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    • pp.186-201
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    • 1995
  • In order to elucidate the fate of the residues of the pyrethroid acaricide-insecticide, acrinathrin in soil, maize plants were grown for one month on the specially-made pots filled with two different types of soils containing fresh and one-month-aged residues of [$^{14}C$]acrinathrin, respectively. The mineralization of [$^{14}C$]acrinathrin to $^{14}CO_2$ during the one-month period of aging and of maize cultivation amounted to $23{\sim}24%$ and $24{\sim}33%$, respectively, of the original $^{14}C$ activities. At harvest after one-month growing, the shoots and roots contained less than 0.1% and 1% of the originally applied $^{14}C$ activity, respectively, whereas the $^{14}C$ activity remaining in soil was $65{\sim}80%$ in both soils. Three degradation products with m/z 198(3-phenoxybenzaldehyde), m/z 214(3-phenoxybenzoic acid), and m/z 228(methyl 3-phenoxybenzoate) besides an unknown were identified from acetone extracts of both soils without and with maize plants after treatment of [$^{14}C$]acrinathrin, by autoradiography and GC-MS, and those with m/z 225(3-phenoxybenzaldehyde cyanohydrin) and m/z 198 (3-phenoxybenzaldehyde) from acetone extract of the Soil A treated with 50 ppm acrinathrin and grown with maize plants for 30 days were identified by mass spectrometry. These results suggested that the hydrolytic cleavage of the ester linkage adjacent to the $^{14}C$ with a cyano group, forming 3-phenoxybenzaldehyde cyanohydrin. The removal of hydrogen cyanide therefrom leads to the formation of 3-phenoxybenzaldehyde as one of the major products. The subsequent oxidation of the aldehyde to 3-phenoxybenzoic acid, followed by decarboxylation would evolve $^{14}CO_2$. Solvent extractability of the soils where maize plants were grown for 1 month and/or [$^{14}C$]acrinathrin was aged for 1 month was less than 31% of the original $^{14}C$ activity and over 95% of the total $^{14}C$ activity in soil extracts was distributed in the organic phase. Accordingly, acrinathrin turned out to be degraded rapidly in both soils and be bound to soil constituents as well, not being available to crops.

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Activities of the Hydrolytic Enzymes Produced by Plant Pathogenic Fungi, Sclerotium rolfsii, Sclerotinia Sclerotinia and Sclerotiorum, and Helminthosporium sigmoideum var. irregulare (수종의 식물병원균(흰비단병균$\cdot$균핵병균 및 좀검은 균핵병균)이 생산하는 가수분해효소의 활성)

  • Cho B. H.;Kim K.
    • Korean journal of applied entomology
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    • v.16 no.4 s.33
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    • pp.199-208
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    • 1977
  • Activities of various hydrolytic enzymes produced by three plant pathogenic fungi, Sclerotium rolfsii Sacc., Sclerotinia sclerotiorum (Lieb.) deBary and Helminthosporium sigmoideum var. irregulare Crallery et Tullius, were measured. Activties and amounts of the enzymes in mycelia, cultural filtrates, and sclerotia(except of sclerotia of H. sigmoideum var. irregulare) were estimated at various pH levels in order to find out optimal pH for their enzymatic activities. Enzymes such as cellulase (ex), invertase, xylanase, $\beta-amylase$, polymethylgalacturonase, polygalacturonase, phosphatase and protease were estimated. Culture solution for production of enzymes was prepared by adding of 10g, D-glucose, 1.3g $NH_4NO_3,\; 0.5g\; MgSO_4,\;7H_2O,\; and\; 1.0g\; KH_2PO_4$ into 1 liter of potato decoction plus 2ml of micro element solution consisting of 0.2mg. Fe, 0.2mg Zn, and 0.1mg Mn as the sulphates into 1 liter of distilled water. All tested mycelia and cultural filtrates were obtained from the cultures incubarted in previous solution for ten days at $25^{\circ}C$, and sclerotia were harvested from PDA plates of 3. days old, The crude enzyme solutions were prepared according to the method of Miyazaki etal. Ten days after incubation, activities of Cx produced by Scl. sclerotiorum were higher than those of the other fung and each of Cx from three fungi showed different pH optima, such as S. rolfsii and Scl. schlerotiorum in acid side (around pH 3.0), H. sigmoideum var. irregulare in neutral side (around pH 6.3). Invertase activities of S. rolfsii were 20 times higher than those of the other fungi in all samples. All tested fungi, however, showed no significant difference between the enzymatic activities of their cultural filtrate and mycelia and the activities in sclerotia of S. rolfsii and Scl. sclerotiorum were hardly recognized. There were multiple peaks on the xylanase activity curves of three fungi in terms of pH values. High activities of the xylanase were revealed in sclerotia of S. rolfsii and Scl. sclerotiorum, and in mycelia of H. sigmoideum var. irregulare. The highest activities of $\beta-amylase$ were shown both in mycelia and cultural filtrate of H. sigmoideum var. irregulae among the tested fungi, and their optimal pH was 6.2 in both mycelia and cultural filtrate. In the S. rofsii and Sel. sclerotiorum, however, the activities of cultural filtrates were higher than those of the other fungi, and optimal pH was 3.0 and 6.2 for cultural filtrate and both mycelia and sclerotia, respectively. Activities of PMG were high in cultural filtrates of all tested fungi, especially in Scl. sclerotiorum and H. sigmoideum var. irregulare. Mycelia of themalso showed the considerable activities. Optimal pH for enzymatic activities were variable with thekind of fungi or with the samples measured. The highest activities of PG were presented by mycelia of S. rolfsii and Scl. sclerotiorum. $9.l\mu /min.\; and\; 9.5\mu g/min.$, respectively. Optimal pH for activity of PG in mycelia was around 4.5 in S. rolfsii and around 3.0 in Scl. sclerotiorum. Phosphatase of S. rolfsii and Scl. sclerotiorum was more active in acid side (optimal PH3. 5) and that of H. sigmoideum var. irregulare showed one peak each in acid, neutral and alkaline side. But the highest peak was at pH 9.5. Protease of all tested fungi was more active at pH 10.0, especially that of the cultural filtrate of H. sigmoideum var. irregualre.

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(1-3, 1-4)-$\beta$-Glucan and Starch Contents and Their Hydrolytic Enzyme Activities in Developing Barley Kernels (등숙 중인 보리 종실중 (1-3, 1-4)-$\beta$-Glucan과 전분 함량 및 이들의 가수분해효소 활성)

  • 윤성중;박상래;유남희
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.42 no.4
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    • pp.403-409
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    • 1997
  • To obtain information on the accumulation of (1-3, 1-4)-$\beta$-glucans during kernel maturation, (1-3, 1-4)-$\beta$-glucan contents and (1-3, 1-4)-$\beta$-glucanase activities were determined in developing kernels of the two Korean cooking barley varieties, Neulssalbori and Saessalbori. (1-3, 1-4)-$\beta$-Glucan contents in kernels at 5 and 10 days after anthesis(DAA) were very low and the contents increased rapidly in kernels at 15 to 25 DAA. (1-3, 1-4)-$\beta$-Glucan content in kernels at harvest was about 3.5 to 4% of kernel dry matter. (1-3, 1-4)-$\beta$-Glucanase activities were relatively higher in younger kernels but the levels of the activity were very low compared with those in germinating kernels. A significant negative correlation was observed between (1-3, 1-4)-$\beta$-glucan contents and (1-3, 1-4)-$\beta$-glucanase activities. Low levels of (1-3, 1-4)-$\beta$-glucanase activites in kernels at 15 to 30 DAA, however, may indicate that (1-3, 1-4)-$\beta$-glucanases have little effect on the final content of (1-3, 1-4)-$\beta$-glucans in barley kernels. Starch contents and $\alpha$-amylase activities were also determined in developing barley kernels. Starch contents increased rapidly as kernels matured and the content at harvest was about 60% of kernel dry matter. Relativley higher levels of $\alpha$-amylase activities in kernels at the earlier developmental stage decreased rapidly as kernels matured.

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Probiotic Properties of Lactobacillus strains Isolated from Kimchi (김치로부터 분리된 Lactobacillus strains의 probiotic 특성)

  • Choi, Hye Jung;Lim, Bo Ram;Kim, Dong Wan;Kwon, Gi-Seok;Joo, Woo Hong
    • Journal of Life Science
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    • v.24 no.11
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    • pp.1231-1237
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    • 2014
  • The objective of this study was to evaluate the safety and functional properties of four potential probiotic strains isolated from Kimchi, traditional Korean fermented vegetables. Based on being higher tolerance to bile salts and showing higher acid resistance or hydrophobic properties, one Lactobacillus arizonensis strain (BCNU 9032) and three L. brevis strains (BCNU 9037, BCNU 9098 and BCNU 9101) were selected in the screening experiment. All strains can survived up to 99% after 3h culture in pH 2.5 and resistant to 1% bile salts. These strains also showed good antimicrobial activities against a number of food borne pathogens, especially against Escherichia coli and Shigella sonnei. The ability to lower cholesterol levels of L. arizonensis BCNU 9032 and L. brevis 9037 were demonstrated by bile salt hydrolytic activity and cholesterol assimilation tests. Moreover, L. brevis BCNU 9098 and BCNU 9101 showed higher adherence to Caco-2 cells (12.76 and 11.86%, respectively) than Lactobacillus rhamnosus GG, a commercial probiotic strain used worldwide. The results suggest that these strains could be used as probiotics.