• 미생물학회지
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• 제38권1호
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• pp.1-6
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• 2002

전분가공폐수처리장 활성슬러지와 논, 밭 토양에서 전분성분을 분해하는 $\alpha$-amylase를 생성하는 균주들을 분리 선별하였다. 이중 효소활성이 우수한 AIV 19를 형태학적 분석 및 API system 과 지방산분석(MIDI) 방법을 이용한 결과 Bacillus 속으로동정되었다. $\alpha$-amylase 생성능을 향상시키기 위해 nitrosoguanidine (NTG)처리를 한 균주 중 선별된 AIV 1940균주의 효소활성능이 원 균주에 비해 1.8배 향상되었다. 분리균주는 그람양성가균으로 길이가 2.3-5 $\mu$ m폭이 0.8-1$\mu$m이었고, Bioscreen C 시스템을 이용한 균주성장능 평가에서는 전분이 3 g/ι농도에서 성장능이 증가되었다. 전분합성폐수를 이용한 유기물제거실험에서는 초기 CODcr가 4,455 mg/ι인 고농도 합성폐수를 1일 3일 반응 후 각각 40.2% 72.3%의 유기물 제거율을 나타내었다.

• 한국동물학회지
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• 제17권3호
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• pp.117-126
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• 1974

한국산 초파리의 생화학적 집단유전학의 체계를 수립하기 위하여 한국의 4개 지역으로 부터 채집, 사육한 초파리(Drosophila melanogaster) 5계통, 즉 여수, 충주, 제주, 신촌 III 및 신촌 IV의 amylase isozyme을 polyacrylamide gel 박층 전기영동법으로 분석, 검토한 결과는 아래와 같다. 1. 신촌 IV를 제외한 대부분의 초파리 계통의 전기영동상에 단 하나의 $band(Amy^1)$가 나타난 경우가 많은데, 이것은 그들 초파리 계통이 anylase에 관하여 homogeneous하다는 것을 의미한다. 2. 신촌 IV 계통은 영동상에 변동이 많은데 (대부분 $Amy^1,2$와 $Amy^1,4$), 이 계통은 amylase에 관하여 heterogeneous 하다는 것을 의미한다. 3. Amylase 계통 중 $Amy^1$이 가장 보편적으로 분포하고 있다. 4. 잡종실험의 결과는 일정한 결론을 내리기가 매우 곤란하다. 5. 한국의 다른 여러 지역으로 부터도 많은 계통의 초파리를 채집하여 더욱 광범위하게 이 문제를 다룰 계획이다.

• 한국미생물·생명공학회지
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• 제49권1호
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• pp.45-56
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• 2021

Improved amylases were developed from protoplast fusants of two amylase-producing Aspergillus species. Twenty regenerated fusants were screened for amylase production using Remazol Brilliant Blue agar. Crude enzyme extracts produced by solid state fermentation of rice bran were assayed for activity. Three variable factors (temperature, pH and enzyme type) were optimized to increase the amylase activity of the parents and selected fusants using rice bran medium and solid state fermentation. Analysis of this optimization was completed using the Central Composite Design (CCD) of the Response Surface Methodology (RSM). Amylase activity assays conducted at room temperature and 80℃ demonstrated that Aspergillus designates, T5 (920.21 U/ml, 966.67 U/ml), T13 (430 U/ml, 1011.11 U/ml) and T14 (500.63 U/ml, 1012.00 U/ml) all exhibited improved function making them the preferred fusants. Amylases produced from these fusants were observed to be active over the entire pH range evaluated in this study. Fusants T5 and T14 demonstrated optimal activity under acidic and alkaline conditions, respectively. Fusants T13 and T14 produced the most amylase at 72 h while parents TA, TC and fusant T5 produced the most amylase after 96 h of incubation. Response surface methodology examinations revealed that the enzyme from fusant T5 was the optimal enzyme demonstrating the highest activity (1055.17 U/ml) at pH 4 and a temperature of 40℃. This enzyme lost activity with further increases in temperature. Starch hydrolysis using fusant T5 gave the highest yield of glucose (1.6158 g/100 ml). The significant activities of the selected fusants at 28 ± 2℃ and 80℃ and the higher sugar yields from cassava starch hydrolysis over their parental strains indicate that it is possible to improve amylase activity using the protoplast fusion technique.

• Journal of Microbiology and Biotechnology
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• 제7권5호
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• pp.293-298
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• 1997

A 4.6-kb HindIII fragment encompassing the complete 140-kDa ${\alpha}$-amylase gene of Lactobacillus amylovorus B 4540 was cloned into pBR322 by the shot gun method. Southern blotting and restriction mapping for the insert were performed. The recombinant 9.0-kb plasmid, pFML1, conferred ${\alpha}$-amylase activity to E. coli and Lactococcus lactis hosts when introduced by electroporation. SDS-PAGE and zymography confirmed the production of 140-kDa ${\alpha}$-amylase and its proteolytic degradation products with enzyme activity in transformants. Total ${\alpha}$-amylase activity of E. coli $DH5{\alpha}$ cells harboring pFML1 was 1.8 units and most activity was detected from cell pellets. Total enzyme activity of L. lactis subsp. lactis MG1363 transformant was five to ten-fold lower than that of E. coli cell but more than half of the activity was detected in the culture supernatant.

• 한국작물학회지
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• 제45권2호
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• pp.108-111
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• 2000

Osmoconditioning and hardening effects for the seed germination of normal and naturally aged rice seeds were studied through analyzing the total sugars and $\alpha$-amylase activity. The normal seeds which used to have high germination rate accelerated germination with the osmoconditioning at the suboptimal temperature of 17$^{\circ}C$. On the other hand, the aged seeds did not affect germination rate at $25^{\circ}C$, while increased germination rate and accelerated germination a little at 17$^{\circ}C$, Hardening of aged seeds increased germination rate by 10-15% compared with control seeds at both 17 and $25^{\circ}C$ and accelerated germination. Total sugar content and $\alpha$-amylase activity of normal seeds were higher than did aged seeds. The aged seeds with treatment of osmoconditioning and hardening increased total sugar content and $\alpha$--amylase activity, but hardening was more effective than osmoconditioning. The $\alpha$--amylase activity was positively correlated with the total sugar content and germination rate.

• 한국작물학회지
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• 제38권3호
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• pp.275-282
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• 1993

본 연구는 한국 옥수수의 $\alpha$-amylase의 유전자 클로닝을 주된 목표로 하여 수행되었다. 이를 위하여 여러 식물체의 $\alpha$-amylase 염기서열로 부터 잘 보존된 부분을 참고로 oligonucleotlde probe 및 PCR primer를 설계, 합성하고, 옥수수의 유묘로부터 전체 RNA를 분리하여 northern blot analysis를 통하여 확인한 다음, 이로부터 첫 번째 가닥 cDNA를 만든 후, 여기서 얻은 RNA : DNA hybrid를 주형으로 한 polymerase chain reaction을 통하여 길이 가 약 500bp되 는 PCR 산물을 얻었다. 이를 클로닝하기 위해 pUC19을 클로닝 백터로 사용하여 재조합 플라스미드인 $\ulcorner$pZM$\alpha$'$\lrcorner$를 만들었다. 합성 probe를 이용, Southern blot analysis한 결과, $\ulcorner$pZM$\alpha$'$\lrcorner$가 옥수수 mRNA로 부터 증폭된 DNA의 일부분을 갖고 있음을 확인하였으며, 그 길이는 PCR 산물과 같은 500bp가량 되는 것으로 나타났다.

• 대한약리학회지
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• 제19권1호
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• pp.71-76
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• 1983

Aminoglycoside antibiotics are reported to enhance the amylase release from isolated slices of pancreas in vitro and the mode of action of aminoglycosides on amylase release is considered different from those of acetylcholine or cholecystokinin(CCK), i.e., electronmicroscopically intact zymogen granules are appeared in the lumen of pancreatic acini by treatment of aminoglycosides. It is known that atropine blocks the secretagogue effect of acetylcholine, and phenoxybenzamine is reported to block the effects of CCK or its analogue caerulein. Present study was undertaken to investigate the mode of action of aminoglycosides on the amylase release using atropine, phenoxybenzamine and propranolol as a membrane stabilizing agent in slices of chicken pancreas. The results are summarized as follows : 1) Streptomycin and kanamycin increased the amylase release significantly from slices of chicken pancreas. 2) The effect of streptomycin was inhibited by atropine but not by phenoxybenzamine or propranolol. 3) The amylase release by acetylcholine was blocked by atropine tut the effect of cholecystokinin octapeptide(CCK-8) was not influenced by atropine, phenoxybenzamine or propranolol. 4) Pretreatment of streptomycin enhanced the secretagogue effect of acetylcholine or CCK-8. From these results it is suggested that amylase releasing effects of aminoglycosides are mediated in part by cholinergic stimulation and in part by membrane alteration and these effects are enhanced by acetylcholine or cholecystokinin.

• 생명과학회지
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• 제12권5호
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• pp.570-576
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• 2002

천일염에서 분리된 고호염성균인 Hainrcular sp. EH-1 으로부터 amylase 생성을 위한 배양조건을 검토하여 최적조건을 설정하였다. 그 결과는 다음과 같다. 아밀라아제 생산을 위한 배양 조건은 4$0^{\circ}C$, pH 7.5이었으며, 최적 생육배지를 soluble starch 1.5%, yeast extract 1.0%, MgSO$_4$7$H_2O$ 2.0%, KCI 0.1%, NaCl 25%로 조성하여 500 $m\ell$ 삼각 플라스크에 100 $m\ell$ 씩 분주하여 110 rpm/min으로 진탕 배양시켰다. 생육과 효소활성은 9일 배양 후 최대치를 나타내어서, 균의 생육정도와 효소생산 사이에서 증식연관형을 나타내었다.

• 환경위생공학
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• 제23권1호
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• pp.25-31
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• 2008

A bacterial strain, Bacillus megaterium L-49 has been isolated and identified that produces alkaline ${\alpha}$-amylase. The cell is ellipsoidal, about $1.0-1.2{\times}3.0-3.6{\mu}m$ in diameter, Gram-positive, motile, and central partial central. Growth occurs in media containing 7% of NaCl. This strain could utilize D-glucose, lactose, xylose, sucrose, mannose, and maltose, and but it does not utilize D-fructose, and glycogen. Among the various concentrations of saturated ammonium sulfate, the retractation ratio in range of 70 to 100% was about 93%. However, in the case of acetone, it was about 98.7%. EDTA has activating effect and Ca2+ has no effect on alkaline ${\alpha}$-amylase activity. The alkaline ${\alpha}$-amylase has low thermal stability. The optimal temperature for reaction is $50^{\circ}C$. The alkaline ${\alpha}$-amylase activity maintained stabilizing at pH 6-11 and the optimal pH for reaction was 9-10.

• 한국식품과학회지
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• 제40권1호
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• pp.70-75
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• 2008

두 개의 amylase를 호알칼리성 Pseudomonas sp. KFCC 10818의 배양액으로부터 정제하여 그 특성을 조사하였다. 정제된 효소의 분자량은 각각 50 kDa과 75 kDa이었다. 이 효소들의 최적반응온도는 각각 $35^{\circ}C$와 $40^{\circ}C$였으며 50 kDa의 효소는 칼슘이온에 의하여 효소활성이 두 배로 촉진되었다. 이 두 효소는 최적 pH가 6-8 부근이었으며 pH 12의 조건에서도 효소활성을 유지하는 알칼리내성을 나타냈다. Maltooligosaccharide이나 soluble starch로부터 maltose와 maltotriose를 최종 효소반응산물로 생산하였다. 두 amylase는 N-말단 아미노산 서열이 각각 QTVPKTTFV와 DTVPGNAFQ로 분석되었다.