• Preventive Nutrition and Food Science
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• 제7권1호
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• pp.28-32
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• 2002

A Micrococcus sp. producing catalase was isolated from soil, and a commercial-scathe cultivation and purification of catalase were conducted. The maximum catalase activity was about 103 BU/mL obtained after 46 hr of cultivation in a 30 L fermenter containing 2% glucose, 2% peptone, 4% yeast extract, and 0.5% NaCl. Soybean sauce, CSL (corn steep liquor), and yeast extract were also studied as media substitutes in the media 30 L fermenter. The optimum medium components for the production catalase were found to be 2% glucose, 4% soybean sauce, and 16% CSL. In a 18 kL fermenter, the stationary phase in the cell growth and maximum catalase activity (112 BU/mL) were reached after 46 hr of cultivation, which was the same result as in the 30 L fermenter. The catalase activity was purified with over 17 folds in four steps with a 33.6% yield. From 104,250 mg of protein after cell lysis, 1,966 mg of the purified enzyme with a specific activity of 192.7 kBU/mg was obtained. The residual activity with the addition of 10% NaCl exhibited more than 100%. The use of just NaCl produced a higher residual activity than combination of bencol (benzyldimethyl ammoniumchloride) and PG (propyleneglycol).

• Biotechnology and Bioprocess Engineering:BBE
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• 제3권1호
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• pp.44-47
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• 1998

Maximum cellulase production was sought by comparing the activities of the cellulases produced by different Trichoderma reesei strains and Aspergillus niger. Trichoderma reesei Rut-C30 showed higher cellulase activity than other Trichoderma reesei stains and Aspergillus niger that was isolated from soil. By optimizing the cultivation conditions during shake flask culture, higher cellulase production could be achieved. The FP(filter paper) activity of 3.7U/ml and CMCase (Carboxymethylcellulase) activity of 60U/ml were obtained from shake flask culture. When it was grown in 2.5L fermentor, where pH and DO levels are controlled, the enzyme activities were 133.35U/ml (CMCase) and 11.67U/ml(FP), respectively. Ammonium sulfate precipitation method was used to recover enzymes from fermentation broth. The dried cellulase powder showed 3074.9U/g of CMCase activity and 166.7U/g of FP activity with 83.5% CMCase recovery.

• 생명과학회지
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• 제8권5호
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• pp.614-621
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• 1998

열안정성 inulinase를 분비하는 Penicillium sp.를 돼지 감자 서식지의 토양으로부터 분리하여 이 균주의 inulinase를 50%-80% 염석, DEAE-Sephacel column chromatography, Toyopearl HW 65 F column chromatography에 의해 정제하여 inulinase 활성을 가진 단일 band를 얻었다. 이 band의 단백질을 polya-cryamide gel electrophoresis 후 추출하여 inulinase 활성을 가지는 것으로 확인되었다. 이 단백질의 분자량은 SDS-PAGE에 의해 약 77,000 dalton으로 추정되었고, 최종 정제도는 53.3배이었다. 정제된 효소의 효소학적 성질을 조사한 결과, 최적온도는 약 6$0^{\circ}C$이었고, 열 안정성은 30-5$0^{\circ}C$에서 비교적 안정하였다. pH 4에서 가장 높은 활성을 나타냈으며, pH 4-5에서는 비교적 안정했고 염기성에서는 아주 낮은 활성을 나타내었다. 금속이온과 다른 화학물질의 영향을 조사한 결과 1mM의 $MnCl_2$와 $CaCl_2$에 의해 활성이 증가했으며 특히, $MnCl_2$는 1.7배까지 활성을 증가시켰다. 그러나, 1mM의 $CuCl_2$,$HgCl_2$와 1mM EDTA 시에는 활성이 저해되었다. TLC분석결과 모든 산물이 monosaccharide였으므로 이 효소는 exo-acting inu-linase로 추정되었고, inulin에 대한 이 효소의 $K_M$치는 $2.2\times10^{-3}$이었다. 이 효소의 결정된 N-terminal 아미노산 배열은 $NH_2$-X-Glu-Tyr-Thr-Glu-Lys/Leu-Tyr-Arg-Pro이다.

• Journal of Microbiology and Biotechnology
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• 제16권3호
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• pp.408-413
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• 2006

A strain, P821, with phospholipase D activity was isolated from soil and identified as a Streptomyces species. The phospholipase D enzyme was purified from a culture broth of the isolated strain using ammonium sulfate precipitation and DEAE-Sepharose, phenyl-Sepharose, and Superose 12 HR column chromatographies. The purified enzyme exhibited an optimum temperature and pH of $55^{\circ}C$ and 6.0, respectively, in the hydrolysis of phosphatidylcholine and remained stable up to $60^{\circ}C$ within a pH range of 3.5-8.0. The enzyme also catalyzed a transphosphatidylation reaction to produce phosphatidylserine with phosphatidylcholine and serine substrates. The optimum conditions for the transphosphatidylation were $30^{\circ}C$ and pH 5.0, indicating quite different optimum conditions for the hydrolysis and transphosphatidylation reactions. The gene encoding the enzyme was cloned by Southern hybridization and colony hybridization using a DNA probe designed from the conserved regions of other known phospholipase D enzymes. The resulting amino acid sequence was most similar to that of the PLD enzyme from Streptomyces halstedii (89.5%). Therefore, the enzyme was confirmed to be a phospholipase D with potential use in the production of phosphatidylserine.

• 한국미생물·생명공학회지
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• 제15권6호
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• pp.408-413
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• 1987

토양으로부터 생 쌀보리 분해 효소 생산능이 있는 170주의 곰팡이와 4주의 방선균을 분리하여 이중 생 쌀보리 분해 효소 생산능이 가장 높은 No. 281 균주를 선정하여 동정한 결과 Rhizopus sp.로 밝혀졌으며, 밀기울을 기본 배지로 한 효소 생산 조건을 검토한 결과 효소 생산 최적 온도는 3$0^{\circ}C$, pH는 4.5-5.0 이고 배양 시간 5일 후에 최고의 역가를 보였다. 생쌀보리를 이용한 무증자 알코올 발효 실험에서는 분리한 Rhizopus sp. No. 281의 효소를 이용했을 때가 시판 효소보다도 에탄올 생성이 2% 증가된 결과를 나타내었다.

• 약학회지
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• 제37권2호
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• pp.129-135
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• 1993

Serratia sp. Y-4 was isolated from soil. Many characteristics of the strain and optimum cultivation condition for protease production were investigated.,The protease from Serratia sp. Y-4 was purified and studied for the properties of the enzyme. The isolated strain was identified to the genus Serratia. The strain was cultivated in 1%-casein, 0.5%-Na$_{3}$PO$_{4}$.7H$_{2}$O, 0.1%-NaCl, 0.05%-KCI, 0.02%-MgSO$_{4}$.7H$_{2}$O, 0.02%-CaCl$_{2}$.2H$_{2}$O, 0.02%-ZnSO$_{4}$.7H$_{2}$O, 0.02%-MnCl$_{2}$.4H$_{2}$O and 0.5%-soy bean oil at pH 7.0 for 35 hrs. The enzyme was purified about 5.89 fold from the culture broth with 31.1% recovery and 19,613 u/mg through ultrafiltration, ammonium sulfate, DEAE-sephacel and Superose-12 chromatography. The purified enzyme was identified as one band by isoelectric focusing, SDS- and native-PAGE. It has maxium activity at $37^{\circ}C$ and pH 9.0. Molecular weight of it is approx. 50 kD and pl is about 6.70. Its Km value for casein was 20 mg/ml. 5 mM-EDTA, 5mM-SDS, Ag$^{+1}$, Cu$^{+2}$, Hg$^{+2}$ and Pb$^{+2}$ inhibited the enzyme.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 1979년도 춘계학술대회
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• pp.113.2-114
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• 1979

$\beta-Galactosidase$ is an enzyme which catalizes hydrolysis of lactose, a natural substrate, to glucose and galctose and transferring some monosac-charide units to active acceptors as sugar or alcohol. The occurence of $\beta-Galactosidase$ is known in various microorganisms, animals and higher plants and has been studied by many investigatigators. Especially, a great deal of articles for the enzyme of E. coli have been presented in genetic control mechanism and induction-repression effects of proteins, On the other hand, in the dairly products industry, it is important to hydrolyes lactosd which is the principal sugar of milk and milk products. During the last few years, the interest in enzymatic hydrolysis of milk lactose has teen increased, because of the lactose intolerence in large groups of the population. Microbial $\beta-Galactosidases$ are considered potentially most suitable for processing milk to hydrolyse lactose and, in recent years, the immobilized enzyme from yeast has been examined. Howev, most of the microbial $\beta-Gal$ actosidase are intracellular enzymes, except a few fungal $\beta-Gala-$ ctosidases, and extracellular $\beta-Galactosidase$ which may be favorable to industrial applieation is not so well investigated. On this studies, a mold producing a potent extracellular $\beta-Galactosidase$ was isolated from soil and identified as an imperfect fungus, Beauveria bassians. In this strain, both extracellular and intracellular $\beta-Galactosidases$ were produced simultaneously and a great increase of the extracellular production was acheved by improving the cultural conditions. The extracellular enzyme was purified more than 1, 000 times by procedures including Phosphocellulose and Sephadex G-200 chromatographies. Several characteristics of the enzymewas clarified with this preparation. The enzyme has a main subunit of molecular weight of 80, 000 which makes an active aggregate. And at neutral pH range, it has optimum pH for activity and stability. The Km value was determined to be 0.45$\times$10$^{-3}$ M for $o-Nitrophenyl-\beta-Galactoside.$ In any event, it is interesting to sttudy the $\beta-Galactosidase$ of B. bassiana for the mechanism of secretion and conformational structure of enzyme.

• 한국유기농업학회지
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• 제25권2호
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• pp.311-328
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• 2017

두둑과 고랑을 재활용한 한국형 무경운 농업에서 유기물 투입과 관수 효과를 구명하고자 무경운 토양에서 시험을 수행하였다. 1. 토양 미생물상 1회 전량관수 조건에서 대두박 투입 처리구의 토양 세균과 곰팡이 수는 대두박 무 투입구에 비하여 많았다. 그리고 유기질비료 투입량이 표준시비량 66%까지 증가되면 세균과 곰팡이 수는 증가되었으나, 그 이상에서는 세균과 곰팡이 수가 감소되는 경향이었다. 곰팡이/세균 비율은 관수 방법과 관계없이 대두박 투입 처리에서 0.6과 1.1로, 무투입 처리의 0.2와 0.5보다 2배 이상 높았다. 1회 전량 관수 조건에서 유기질 비료 시비량이 증가되면 대두박을 투입한 처리는 방선균 수는 감소되는 경향이었으나, 대두박 무투입에서는 증가되었다. 2회 분할 관수는 1회 전량관수에 비하여 대두박 무 투입 조건에서 세균과 곰팡이 수가 증가되었으나, 대두박 투입조건에서는 방선균 수가 증가되었다. 2. 토양 효소 유기질 비료의 시비량이 증가되면 토양 내 Chitinase 활성은 대두박 투입 토양에서 감소되고, 대두박 무 투입에서는 증가되는 경향이었다. 그러나 대두박을 투입에 관계없이 2회 분할 관수는 1회 전량관수에 비하여 Chitinase 활성이 증가되었다. 1회 전량관수 조건에서 대두박 투입 처리구의 ${\beta}$-Glucosidase 활성은 무투입에 비하여 높았으며, 유기질 비료 투입량이 증가되면 표준시비량의 66%까지는 ${\beta}$-Glucosidase 활성이 증가되었으나, 표준시비량에서는 감소되었다. 대두박 무투입 조건에서 2회 분할관수 토양 내 ${\beta}$-Glucosidase 활성은 1회 전량관수에 비하여 현저하게 증가되었다. 1회 전량관수 조건에서 대두박을 투입한 처리의 N-acetyl-${\beta}$-D-glucosaminidase의 활성은 무투입구에 비하여 높았다. 대두박 투입 처리에서 유기질 비료 투입량이 표준시비량의 66%까지 증가되면 N-acetyl-${\beta}$-D-glucosaminidase의 활성은 증가되었으나, 표준시비량에서는 유의적인 차이가 없었다. 대두박 무투입 조건에서 2회 분할관수는 1회 전량관수에 비하여 N-acetyl-${\beta}$-D-glucosaminidase의 활성은 증가되었다. 대두박 무투입 조건에서 유기질 비료 시비량이 표준량의 66% 수준에서는 토양 내 산성인산가수분해효소(Acid phosphatase)의 활성 높았다. 대두박 투입 조건에서는 유기질 비료 시비량이 증가되면 산성인산가수분해효소(Acid phosphatase)의 활성은 증가되는 경향이었다. 3. 토양 AMF 대두박 무투입 조건에서 유기질 비료의 투입량이 표준시비량의 66%까지 증가되면 토양의 내생균근균의(AMF) 포자수는 증가되었으나, 유기질 비료 투입량이 표준시비량에서는 근균의 포자수는 감소되었다. 그러나 대두박 투입에서 근균의 포자수는 유기질 비료 투입량에 따른 유의적인 차이가 없었다. 그리고 내생 근균의 고추 뿌리에 정착률은 대두박 투입량에 따른 유의적인 차이가 없었으며, 2회 분할 관수도 같은 경향이었다.

• 한국지하수토양환경학회:학술대회논문집
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• 한국지하수토양환경학회 2003년도 총회 및 춘계학술발표회
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• pp.188-191
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• 2003

Sing]e-well-push-pull tests were developed for use in assessing the feasibility of in-situ aerobic cometabolism of chlorinated aliphatic hydrocarbons (CAHs). The series includes Transport tests, Biostimulation tests, and Activity tests. Transport tests are conducted to evaluate the mobility of solutes used in subsequent tests. These included bromide or chloride (conservative tracers), propane (growth substrate), ethylene, propylene (CAH surrogates), dissolved oxygen (electron acceptor) and nitrate (a minor nutrient). Tests were conducted at an experimental well field of Oregon State University. At this site, extraction phase breakthrough curves for all solutes were similar, indicating apparent conservative transport of the dissolved gases and nitrate prior to biostimulation. Biostimulation tests were conducted to stimulate propane-utilizing activity of indigenous microorganisms and consisted of sequential injections of site groundwater containing dissolved propane and oxygen. Biostimulation was detected by the increase in rates of propane and oxygen utilization after each injection. Activity tests were conducted to quantify rates of substrate utilization and to confirm that CAH-transforming activity had been stimulated. In particular, the transformation of injected CAH surrogates ethylene and propylene to the cometabolic byproducts ethylene oxide and propylene oxide provided evidence that activity of the monooxygenase enzyme system, responsible for aerobic cometabolic transformations of CAHs had been stimulated. Estimated zero-order transformation rates decreased in the order propane > ethylene > propylene. The series of push-pu3l tests developed and field tested in this study should prove useful for conducting rapid, low-cost feasibility assessments for in situ aerobic cometabolism of CAHs.

• Journal of the Korean Wood Science and Technology
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• 제29권3호
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• pp.112-122
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• 2001

Laccase enzymes from Cerrena unicolor and Trametes versicolor were immobilized on the activated glass beads (CPG), silica gel (SG) and soil (SL). The heterogeneous matrices were activated by ${\gamma}$-aminopropyltriethoxysilane (APTES) and glutaraldehyde (GA), and their surfaces were coated by keratin (KER) on activated or non-activated CPG, SG and SL. The laccase activities were tested in the aqueous solution for the native and immobilized preparations using different pH and temperature conditions. By keratin coating on supports, in the cases of CPG-KER and SL-KER, the immobilization yield was increased from about 80% to 90%. Moreover, much less protein was immobilized in keratin coated matrices than in inorganic ones alone (e.g. on CPG-KER 57.6%, whereas on CPG alone 80.6%). Laccase immobilization on keratin coated inorganic matrices was generally more effective than that of non-coated matrices. Concerned to pH dependency, the optima pH for immobilized laccases generally shifted towards to higher values, 5.5-5.8 and even 5.9 in the case of keratin for C. unicolor and from 5.3 to 5.7 for T. versicolor, respectively, and decreased less gradually both in acidic and alkaline regions. The immobilized laccase was more stable against thermal denaturation. This seems particularly true at $75^{\circ}C$ in the case of C. unicolor, where the activity of immobilized enzyme is > 50% higher than that of the free enzyme. For T. versicolor the respective values were $65^{\circ}C$, and 50%.