• Title/Summary/Keyword: soil enzymes

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Properties of Chorismate Mutase from intrasporangium sp. (Intrasporangium속 방선균의 Chorismate Mutase 성질)

  • 조원대;신광순;최용진;양한철
    • Microbiology and Biotechnology Letters
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    • v.16 no.4
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    • pp.310-315
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    • 1988
  • Two isoenzymes of chorismate mutase(E.C.5.4.99.5) designated as chorismate mutase I(CM I) and chorismate mutase II(CM II), were detected and partially purified from a sp. of intrasporangium isolated from soil. CM I and CM II had pH optima of pH 6.5 and 8.0, respectively and showed the same temperature optimum of 45$^{\circ}C$. The activation energy of the enzymatic reaction was estimated to be 14.7kcal/ mole with CM I and 10.8kcal/mole with CM II. The affinity of isoenzyme CM I for substrate(Km= 1.35mM) was almost the same level as that of CM II(Km = 1.22mM). Both isoenzymes were stable at pH values ranged from pH 6.5 to 9.0, but rapidly denaturated at temperatures above 45$^{\circ}C$. CM II was activated about 7$^{\circ}C$ of its activity by $Ba^{++}$ or $Mg^{++}$ while CM I was slightly inhibited by the same metal ions. Thiol compounds were found not to be necessary for stability of the two enzymes but Co$^{++}$ and EDTA had a little stabilizing effect on CM II only. p-Chloromercuribenzoate strongly inactivated the activities of both enzymes but the reducing agents such as dithiothreitol and L-cysteine protected them against the pCMB inhibition.

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Characterization of Nitrile-hydrolyzing Enzymes Produced from Rhodococcus erythropolis (니트릴 분해효소 생산균인 Rhodococcus erythropolis의 발굴 및 효소 특성 연구)

  • Park Hyo-Jung;Park Ha-Joo;Uhm Ki-Nam;Kim Hyung-Kwoun
    • Microbiology and Biotechnology Letters
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    • v.34 no.3
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    • pp.204-210
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    • 2006
  • Ethyl (S)-4-chloro-3-hydroxybutyrate is a useful intermediate for the synthesis of Atorvastatin, a chiral drug to hypercholesterolemia. In this research, two 4-chloro-3-hydroxybutyro-nitrile-degrading strains were isolated from soil sample. They were identified as Rhodococcus erythropolis strains by 16S rRNA analysis. The nitrile-degrading enzyme(s) were suggested to be nitrile hydratase and amidase rather than nitrilase from the result of thin layer chromatography analysis. The corresponding genes were obtained by PCR cloning method. The predicted protein sequences had identities more than 96% with nitrile hydratase ${\alpha}-subunit$, nitrile hydratase ${\beta}-subunit$, and amidase of R. erythropolis. The 4-chloro-3-hydroxybutyronitrile-hydrolyzing activities in both strains were increased dramatically by ${\varepsilon}-caprolactam$ which was known as good inducer for nitrile hydratase. Both intact cells and cell-free extract could hydrolyze the nitrile compound. So, the intact cell and the enzymes could be used as potential biocatalyst for the production of 4-chloro-3-hydroxybutyric acid.

Effects of Hypoxia on Root Growth and Anaerobic Fermentative Enzymes in Winter Cereal Seedlings (저산소 조건하에서 맥류 유묘의 근생장 및 혐기발효 효소의 반응)

  • Park Myoung Ryoul;Lim Jeong Hyun;Yoo Nam Hee;Kwon In Sook;Kim Jung Gon;Choi Kyung Gu;Yun Song Joong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.50 no.6
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    • pp.400-405
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    • 2005
  • Wet-injury often occurs in upland cereals growing in the paddy field due to oxygen deficiency in the rhizosphere caused by excessive water in the soil. Under hypoxia, energy metabolism is diminished causing non­reversible damage to root cells. This study was conducted to investigate effects of hypoxia on root growth and enzymes involved in the fermentative energy metabolism in upland cereals including barley, wheat, rye and triticale. Young seedlings were subject to hypoxia for up to 7 days. Root fresh weight and dry weight were decreased significantly by hypoxia for 5 to 7 days in all cereal seedlings. Root growth retardation under hypoxia was lowest in barley. Hypoxia-induced increases in activity and isozyme expression of alcohol dehydrogenase (ADH) and lactate dehydrogenase (LDH) were commonly observed in roots of all cereal seedlings. The inherent ADH activity levels were higher in barley but the hypoxia-induced increases in ADH activities were lowest in barley than other cereals. The inherent LDH activity levels were lower in barley and the hypoxia-induced increases in LDH activities were lower in barley than other cereals. The results suggest the importance of the rapid enhancement of fermentative enzyme systems for increased tolerance to hypoxia.

Importance of Extracellular Enzyme Activities in Northern Peatland Biogeochemistry-Possible Coupling with Trace Gas Emission and DOC Dynamics (북구 이탄습지의 생지화학적 반응에 있어서 체외효소의 중요성-미량기체 발생량 및 용존유기탄소 동태와의 연관성에 대하여)

  • Freeman, Chris;Park, Seok-Soon;Kang, Ho-Jeong
    • Korean Journal of Ecology and Environment
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    • v.39 no.4 s.118
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    • pp.435-444
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    • 2006
  • A suite of extracellular enzyme activities involved in organic carbon decomposition were determined in three northern peatlands (a bog, a fen, and a swamp) over a 12 month period along with trace gas ($CO_2$ and $N_2O$) flux and DOC dynamics in the wetlands. The activities varied $0.008-0.066\;{\mu}mole\;g^{-1}\;min^{-1}$, $0.003-0.021\;{\mu}mole\;g^{-1}\;min^{-1}$, $0.003-0.016\;{\mu}mole\;g^{-1}\;min^{-1}$, $0.004-0.047\;{\mu}mole\;g^{-1}\;min^{-1}$, for ${\beta}-glucosidase$, cellobiohydrolase, ${\beta}-xylosidase$, and N-acetylglucosaminidase, respectively. In general, the activities were highest in the forested swamp followed by the fen and the bog. When the data from three wetlands are combined, the enzyme activities exhibited significant positive correlations with trace gas emission and available carbon. Further, the average activity of 4 enzymes explained about 20-40% of the variations of trace gas emssion and available carbon. The results indicate that enzymes related to the mineralization of organic carbon may play an important role in trace gas flux and DOC dynamics in northern peatlands.

Protective Mechanism of Salidroside Isolated from Acer termentosum Max on Cyclosporine-Induced Nephrotoxicity in Rats (흰쥐에서 Cyclosporine 유도가 산겨릅나무에서 분리한 salidroside의 신독성 경감기전에 관한 효과)

  • Kim, Sung-Hoon;Park, Hee-Juhn;Choi, Jong-Won
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.1
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    • pp.166-173
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    • 2009
  • Cyclosporine(CsA) is an immunosupressant drug widely used in post-allogeneic organ transplant to reduce the activity of the patient's immune system and so the risk of organ rejection. It has been studied in transplants of skin, heart, kidney, liver, lung, pancreas, bone marrow and small intestine. Initially isolated from a Norwegian soil sample, Both kidney and liver dysfunction are prominent side effects of CsA. The present study was designed to determine the possible protective effect of salidroside(Sal) isolated from the BuOH extract of Acer termentosum Max against oxidative damage in CsA-treated(50 mg/kg, ip) nephrotoxicity rats. Results showed oral administration of methanol and butanol extact of Acer termentosum Max(200 mg/kg, po) significantly reduced activities of marker enzymes(BUN, Creatinine) and LDH activity in serum to CsA induced experimental kidney injured rats. And significantly decrcease of protein amount level in urine and activities of free radical formation enzyme were significantly improved by the treatment of Sal. And significantly decrcease of MDA level in kidney and activities of calalase, glutathione peroxidation and SOD were significantly improved by the treatment of Sal(20 mg/kg, po). But glutathione concentration and glutathione S-transferase actitity was not affected. Results of this study revealed that Sal could afford a significant protection in the alleviation of CsA-induced nephrotoxicity injury.

R-Stereoselective Amidase from Rhodococcus erythropolis No. 7 Acting on 4-Chloro-3-Hydroxybutyramide

  • Park, Ha-Ju;Uhm, Ki-Nam;Kim, Hyung-Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.552-559
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    • 2008
  • Ethyl (S)-4-chloro-3-hydroxybutyrate is an intermediate for the synthesis of Atorvastatin, a chiral drug used for hypercholesterolemia. A Rhodococcus erythropolisstrain (No.7) able to convert 4-chloro-3-hydroxybutyronitrile into 4-chloro-3-hydroxybutyric acid has recently been isolated from soil. This activity has been regarded as having been caused by the successive actions of the nitrile hydratase and amidase. In this instance, the corresponding amidase gene was cloned from the R. erythropolis strain and expressed in Escherichia coli cells. A soluble active form of amidase enzyme was obtained at $18^{\circ}C$. The Ni column-purified recombinant amidase was found to have a specific activity of 3.89 U/mg toward the substrate isobutyramide. The amidase was found to exhibit a higher degree of activity when used with mid-chain substrates than with short-chain ones. Put differently, amongst the various amides tested, isobutyramide and butyramide were found to be hydrolyzed the most rapidly. In addition to amidase activity, the enzyme was found to exhibit acyltransferase activity when hydroxyl amine was present. This dual activity has also been observed in other enzymes belonging to the same amidase group (E.C. 3.5.1.4). Moreover, the purified enzyme was proven to be able to enantioselectively hydrolyze 4-chloro-3-hydroxybutyramide into the corresponding acid. The e.e. value was measured to be 52% when the conversion yield was 57%. Although this e.e. value is low for direct commercial use, molecular evolution could eventually result in this amidase being used as a biocatalyst for the production of ethyl (S)-4-chloro-3-hydroxybutyrate.

STRATEGIES TO REDUCE ENVIRONMENTAL POLLUTION FROM ANIMAL MANURE: PRINCIPLES AND NUTRITIONAL MANAGEMENT - A REVIEW -

  • Paik, I.K.;Blair, Robert;Jacob, Jacqueline
    • Asian-Australasian Journal of Animal Sciences
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    • v.9 no.6
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    • pp.615-635
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    • 1996
  • The animal industry must be environmentally sound to ensure its long-term sustainable growth. Livestock wastes mostly manure, can be a valuable resource as well as a potential hazard to environment. The first option of manure management is developing an 'environmentally sound' feeding program and feeds so there are less excreted nutrients that need to be managed. Once the manure is produced it can be best utilized as a fertilizer of a soil conditioner. In many countries the amount of manure that can be spread on land depends on the nutrient requirements of the crop being grown. The laws specify maximum application rates and not animal stocking rates. Farmer who reduce the N and P component of manure can release pressure on the environment without having to reduce the number of animals. There are alternative system for housing and manure treatment which generate manure that are easier to handle and have less pollutants or more economic value. Treated animal waste may also be used as a feedstuff or fuel source. Most of the options of waste management result in increased costs to implement. It is necessary to assess the economics in order to find an acceptable compromise between the increased costs and the benefit to the environment. Animal welfare is also becoming more and more of an issue and it will lead to systems where animals are kept in less confined environment. The new system will have a great impact in the waste management system in the future.

Keratinase Production by Recalcitrant Feather Degrading Pseudomonas Geniculata and Its Plant Growth Promoting Activity (난분해성 우모분해 Pseudomonas geniculata에 의한 케라틴 분해효소 생산 및 식물성장 촉진 활성)

  • Go, Tae-Hun;Lee, Sang-Mee;Lee, Na-Ri;Jeong, Seong-Yun;Hong, Chang-Oh;Son, Hong-Joo
    • Journal of Environmental Science International
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    • v.22 no.11
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    • pp.1457-1464
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    • 2013
  • We investigated the optimal conditions for keratinase production by feather-degrading Pseudomonas geniculata H10 using one variable at a time (OVT) method. The optimal medium composition and cultural condition for keratinase production were determined to be glucose 0.15% (w/v), beef extract 0.08% (w/v), $KH_2PO_4$ 0.12% (w/v), $K_2HPO_4$ 0.02% (w/v), NaCl 0.07% (w/v), $MgSO_4{\cdot}7H_2O$ 0.03%, $MgCl_2{\cdot}6H_2O$ 0.04% along with initial pH 10 at 200 rpm and $25^{\circ}C$, respectively. The production yield of keratinase was 31.6 U/ml in an optimal condition, showing 4.6-fold higher than that in basal medium. The strain H10 also showed plant growth promoting activities. This strain had ammonification activity and produced indoleacetic acid (IAA), siderophore and a variety of hydrolytic enzymes such as protease, lipase and chitinase. Therefore, this study showed that P. geniculata H10 could be not only used to upgrade the nutritional value of feather wastes but also useful in situ biodegradation of feather wastes. Moreover, it is also a potential candidate for the development of biofertilizing agent applicable to crop plant soil.

Alleviation of Salt Stress in Pepper (Capsicum annum L.) Plants by Plant Growth-Promoting Rhizobacteria

  • Hahm, Mi-Seon;Son, Jin-Soo;Hwang, Ye-Ji;Kwon, Duk-Kee;Ghim, Sa-Youl
    • Journal of Microbiology and Biotechnology
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    • v.27 no.10
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    • pp.1790-1797
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    • 2017
  • In the present study, we demonstrate that the growth of salt-stressed pepper plants is improved by inoculation with plant growth-promoting rhizobacteria (PGPR). Three PGPR strains (Microbacterium oleivorans KNUC7074, Brevibacterium iodinum KNUC7183, and Rhizobium massiliae KNUC7586) were isolated from the rhizosphere of pepper plants growing in saline soil, and pepper plants inoculated with these PGPR strains exhibited significantly greater plant height, fresh weight, dry weight, and total chlorophyll content than non-inoculated plants. In addition, salt-stressed pepper plants that were inoculated with B. iodinum KNUC7183 and R. massiliae KNUC7586 possessed significantly different total soluble sugar and proline contents from non-inoculated controls, and the activity of several antioxidant enzymes (ascorbate peroxidase, guaiacol peroxidase, and catalase) was also elevated in PGPR-treated plants under salt stress. Overall, these results suggest that the inoculation of pepper plants with M. oleivorans KNUC7074, B. iodinum KNUC7183, and R. massiliae KNUC7586 can alleviate the harmful effects of salt stress on plant growth.

Investigation of gene encoding catechol 1,2-dioxygenase from Phenol-degrading, Rhodococcus sp. EL-GT

  • Lee, Hui-Jeong;Han, Chang-Min;Jo, Sun-Ja;Park, Geun-Tae;Park, Jae-Rim;Lee, Sang-Jun
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.549-551
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    • 2001
  • The heavy use of petroleum products in modern livings has brought ubiquitous environmental contaminants of aromatic compounds, which persist in aquatic and geo-environment without the substantial degradation. The persistence and accumulation of the aromatic compounds, which include xylene, phenol, toluene, phthalate, and so on are known to cause serious problems in our environments. Some of soil and aquatic microorganisms facilitate their growth by degrading aromatic compounds and utilizing degrading products as growth substrates, the biodegradation helps the reentry of carbons of aromatic compounds, preventing their accumulation in our environments. The metabolic studies on the degradation of aromatic compounds by microoganisms were extensively carried out along with their genetic studies. A Rhodococcus sp. EL-GT isolated in activated sludges has shown the excellent ability to grow on phenol as a sole carbon source. In the present study investigated a gene encoding phenol-degrading enzymes from a Rhodococcus sp. EL-GT.

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