• 한국작물학회지
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• 제34권s02호
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• pp.66-80
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• 1989

Salt injury in rice is caused mainly by the salinity in soil and in the irrigated water, and occasionaly by salinity delivered through typhoon from the sea. The salt concentration of rice plants increased with higher salinity in the soil of the rice growing. The climatic conditions, high temperature and solar radiation and dry conditions promote the salt absorption of rice plant in saline soil. The higher salt accumulation in the rice plant generally reduces the root activity and inhibits the absorption of minerals of rice plant, resulting the reduction of photosynthesis. The salt damages of rice plant, however, are different from different growth stage of rice plants as follows: 1. Germination of rice seed was slightly delayed up to 1.0％ of salt concentration and remarkably at 1. 5％, but none of rice seeds were germinated at 2.5％. This may be due to the delayed water uptake of rice seeds and the inhibition of enzyme activity, 2. It was enable to establish rice seedlings at seed bed by 0.2％ of salt concentration with some reduction of leaf elongation. The increasing of 0.3％ salt concentration caused to the seedling death with varietal differences, but most of seedlings were death at 0.4％ with no varietal differences. 3. Seedlings grown at the nursery over 0.1％ salt, gradually reduced in rooting activity after transplanting according to increasing the salt concentration from 0.1％ up to 0.3％ of paddy field. However, the seedlings grown in normal seed bed showed no difference in rooting between varieties up to 0.1％ but significantly different at 0.3％ between varieties, but greatly reduced at 0.5％ and died at last in paddy after transplanting. 4. At panicle initiation stage, rice plant delayed in heading by salt damage, at meiotic stage reduced in grains and its filling rate due to inhibition of glume and pollen developing, and salt damage at heading stage and till 3 weeks after heading caused to reduction of fertilization and ripening rate. In viewpoint of agricultural policy the overcoming strategy for salt injury is to secure sufficient water source. Irrigation and drainage systems as well as underground drainage is necessary to desalinize more effectively. This must be the most effective and positive way except cost. By cultural practice, growing the salt tolerant variety with high population could increase yield. The intermittent irrigation and fresh water flooding especially at transplanting and from panicle initiation to heading stage, the most sensitive to salt injury, is important to reduce the salt content in saline soil. During the off-cropping season, plough and rotavation with flooding followed by drainage, or submersion and drainage with groove could improve the desalinization. Increase of nitrogen fertilizer with more split application, and soil improvement by lime, organic matter and forign soil addition, could increase the rice yield. Shift of trans-planting is one of the way to escape from the salt injury.

• Journal of Microbiology and Biotechnology
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• 제16권12호
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• pp.1868-1873
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• 2006

A novel microorganism that could degrade high molecular weight gellan was screened and isolated from soil. On gellan plate, the microorganism grew well and completely liquefied the plate. The gellan-degrading microorganism was isolated by pure culture on glucose and nutrient agar medium afterwards. The 16S rDNA sequence analysis and biochemical tests using an API 50CHB/20E kit revealed that the strain belonged to Bacillus sp. The isolate, named as Bacillus sp. YJ-1, showed optimum gellan-degrading activity in 0.5% gellan medium at pH 7.5 and 37$^{\circ}C$. The activity was measured and evaluated by the thiobarbituric acid and thin-layer chromatography method. Mass spectrometry revealed that the major gellan.. depolymerized product was an unsaturated tetrasaccharide consisting of $\Delta$4,5-glucuronic acid-(1$\rightarrow$4 )-$\beta$-D-glucose-(1$\rightarrow$4)- $\alpha$-L-rhamnose-(1$\rightarrow$3)-$\beta$-D-glucose, which is a dehydrated repeating unit of gellan, thus the enzyme was identified as gellan lyase. When the gellan was present in the medium, the gellan-degrading activity was much higher than that in glucose-grown cells. These results indicate that in the presence of gellan, Bacillus sp. YJ-1 is able to metabolize the gellan by inducing gellan-degrading enzymes that can degrade gellan into small molecular weight oligosaccharides, and then the gellan. depolymerized products are taken up by the cells and utilized by intracellular enzymes.

• 한국미생물·생명공학회지
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• 제31권1호
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• pp.57-62
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• 2003

토양으로부터 $\beta$-mannanase활성이 우수한 균주를 분리하여 형태학적, 생화학적 동정과정을 거쳐 Bacillus subtilis JS-1으로 동정하였다. 분리균이 생산하는 $\beta$-mannanase 효소의 최적활성은 55$^{\circ}C$와 pH 5.0이었다. 탄소원이 다른 배지에서 배양한 분리 균주의 상등액을 전기영동하여 효소활성을 관찰한 결과 탄소원에 상관없이 분자량 130kDa에 해당하는 단일 단백질만이 효소 활성을 나타내었다 Bacillus subtilis JS-1은 탄소원으로 lactose와 locust bean gum이 존재할 때 $\beta$-mannanase 생산성이 크게 증가하는 것으로 나타났으며, lactose와 locust bean gum이 각각 0.5 % 존재할 때 배양 상등액의 $\beta$-mannanase 활성은 30U/ml과 45U/ml로 탄소원이 없는 대조구에 비해 최대 18배 정도 생산성이 증가하였다. 배지에 locust bean gum을 첨가하였을 때 효소 생산성 뿐만 아니라 균체의 성장도 함께 증가하는 것으로 보아 분리균주는 locust bean gum을 분해하여 에너지원으로 이용하는 것으로 판단된다

• Journal of Microbiology and Biotechnology
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• 제13권6호
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• pp.960-968
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• 2003

A bacterial isolate showing a strong endochitosanase activity was isolated from soil and then characterized. The isolate was identified and designated as Bacillus cereus P16, based on morphological and biochemical properties, assimilation tests, cellular fatty acids pattern, along with 16S rRNA gene sequence. The optimized medium for producing extracellular chitosanase in a batch culture contained 1% tryptone, 0.5% chitosan, and 1% NaCl (pH 7.0). Powder chitosan and tryptone served the best as carbon and nitrogen sources, respectively, for the chitosanase production. Chitosanase activity was the highest when culture was completed at $37^{\circ}C$ among various temperatures ($20-42^{\circ}C$) tested in a shaking incubator (200 rpm). The levels of chitosanase activity in the culture fluid were 2.0 U/ml and 3.8 U/ml, respectively, when incubated in a flask for 60 h and in a jar fermenter for 24 h. The culture supernatant showed a strong liquefying activity on the soluble chitosan. The viscosity of 1% chitosan solution, that was incubated with the culture supernatant, was rapidly decreased, suggesting the secretion of endochitosanolytic enzymes by P16. The culture fluid revealed six endo-type chitosanase isozymes, two major (38 and 45 kD), and four minor (54, 65, 82, and 96 kD) forms by staining profile. The crude enzymes were very stable, and full activity was maintained for 4 weeks at $4^{\circ}C\;or\;-20^{\circ}C$ in the culture supernatant, suggesting a highly desirable stability rate for making an industrial application of the crude enzymes. The supernatant also cleaved the insoluble chitosan powder, but the hydrolysis rate was much lower. The enzymic degradation products of chitosan contained $(GlcN)_n$ (n=2-8). The concentration of chitosan in the reaction mixture of the crude enzyme affected the chitooligosaccharides composition of the hydrolysis products. When the higher concentration of chitosan was used, the higher degree of polymerized chitooligosaccharides were produced. By comparison with other commercial chitosanase preparations, P16 was indeed found to be a valuable enzyme source for industrial production of chitooligosaccharides from chitosan.

• Journal of the Korean Wood Science and Technology
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• 제48권3호
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• pp.376-392
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• 2020

우리나라는 울릉도 및 제주도를 포함한 전국에서 흰개미에 의한 목조 문화재의 피해가 보고되어 있다. 흰개미에 의한 피해를 감소시키고자 훈증처리나 토양에 약물을 처리하여 살충 및 방충을 하는 것이 대부분이며 비용과 안전성의 문제로 인해 점점 처리하는 횟수가 감소하는 추세이다. 이런 상황을 대처하기 위해 새로운 방법이 필요한 실정이므로 전남 신안에서 채집한 흰개미에서 일개미만 선별하여 효소를 추출한 후 목재 구성성분인 cellulose와 hemicellulose의 xylan을 기질로 하여 효소 활성을 측정한 결과, 분자량이 큰 cellulose 보다 xylan에서 흰개미 장내 효소의 활성이 크게 나타났다. 그러므로 본 연구에서는 xylan을 기질로 하여 흰개미 장내 효소의 활성을 억제하는 약용식물 추출물 600여 종을 탐색한 결과, 용뇌, 마황, 박하뇌에서 억제효과가 크게 나타났다. 선별된 용뇌(Borneolum Syntheticum), 마황(Ephedra sinica), 박하뇌(Menthol) 추출물은 흰개미 장내 효소의 활성을 억제할 뿐만 아니라 직접 처리한 결과에서도 살충활성 및 섭식 저해 효과가 있음을 확인하였다.

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2001년도 제32회 학술심포지움
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• pp.67-86
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• 2001

A strain producing strongly fibrinolytic enzyme was isolated from soil and was identified to be Bacillus subtilis by biochemical and physiological characterization. The optimal culture conditions for the production of fibrinolytic enzyme was determined to be 1.0% tryptone, 1.5% soluble starch, 0.5% Peptone, 0.5% NaCl, $(NH_{4})_{3}PO_4.3H_{2}O, and MgSO_{4}.7H_{2}O.$ Initial pH and temperature were pH 8.0 and $30^{\circ}C$ , respectively, The highest enzyme production was observed at 30 hours of cultivation at $30^{\circ}C$ The fibrinolytic enzyme was purified to homogeneity by DEAE Sephadex A-50 ion exchange column chromatography, 70% ammonium sulfate precipitation, Sephadex G-200 and G-75 gel filtration column chromatography. The molecular weight of the purified enzyme was 28,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A gene encoding the fibrinolytic enzyme was cloned into a plasmid vector pBluescript, transforming E.coli XL-1 Blue. The clone was able to degrade fibrin, This indicated that the gene could encode a fibrinolytic enzyme. The nucleotide sequence of the 2.7 kb insert was determined in both direction. One open reading frame composed of 1023 nucleotides was found to be a potential protein coding region. There was the putative Shine-Dalgano sequence and TATA box upstream of the open reading frame. The homology search data in the genome database showed that both the 2.7 kb insert and 1 kb open reading frame carried no significance in the nucleotide sequence of known fibrinolytic enzyme from Bacillus serovars. The recombinant cell harboring the novel gene involved in fibrinolysis was subjected to protein purification. The molecular mass of the purified fibrinolytic enzyme was determined to be 31864 Dalton, which was highly in accordance with the molecular mass(33 kDa) of the fibrinolytic gene deduced from the insert. The fibrinolytic enzyme was Purified 50.5 folds to homogeneity in overall yield of 10.7% by DEAE Sephadex A-50 ion exchange, 85% ammonium sulfate precipitation, Sephadex G-50, Superdex 75 HR FPLC gel filtration. In conclusion, a novel fibrinolytic gene from Bacillus subtilis was identified and characterized by cloning a genomic library of Bacillus subtilis into pBleuscript. For the soybean fermented by this strain, it is found that there increased assistant protein about 20% compared to the soybean not fermented and increased about 30% according to amino acid analysis and, in particular, essential amino acid increased about 40%. When keeping this fermented soybean powder at room temperature for about 70days, it showed very high stability maintaining almost perfect activity and, therefore, it gave us great suggestion its possibility of development as a new functional food.

• 한국미생물·생명공학회지
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• 제17권6호
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• pp.580-586
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• 1989

Penicillium sp. CB-20 이 생성하는 polygalacturonase의 최대 효소활성을 위한 pH는 5.0, 최적온도는 4$0^{\circ}C$였으며, 이 효소는 약산성의 pH에서 안정성을 보였고, 온도에 의한 안정성은 3$0^{\circ}C$였으며 4$0^{\circ}C$이상에서는 급격한 효소단백질의 불활성화가 진행되었다. 금속이온 중 $Mg^{++}$, $Zn^{++}$, $Ba^{++}$ 등이 활성을 촉진시켰고, Ag$^{+}$, Cu$^{++}$, Pb$^{++}$, Fe$^{+++}$, $Ca^{++}$, $Na^{++}$, Mnc 등은 효소활성을 저해하였으며, 본 균주가 생성하는 호소의 $K_m$값과 V$_{max}$값, 활성화에너지는 2.13$\times$$10^{-2}$mol/$\ell$, 104.17 $\mu$mol/min, 2,499 Kcal/mol 이었으며, pectin보다 polygalacturonic acid를 특이 적으로 분해하였다. 효소저해제 중 EDTA, DNP, $H_2O$$_2$ 등에 의해 효소활성이 제해되어 효소분자 중의 금속이 활성에 관여하며, 말단 아미노기와 histidine의 imidazole기가 효소활성에 관여함이 입증되었던 반면에 효소분자 중 SH기는 활성에 관여하지 않음을 알 수 있었다. 이 효소의 기질분해 양상을 종이 크로마토그라피로 확인한 결과 반응초기에는 monomer, dimer oligomer 등이 생성되었고 시간이 경과할수록 oligomer는 사라지고 monomer dimer 만이 생성되는 것으로 보아 endo 형인 것으로 판단되었으며, 효소의 과즙청징도 측정에서는 약 8 unit 에서 거의 청징화되었다.

• 한국식품영양과학회지
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• 제21권2호
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• pp.195-200
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• 1992

Rhizopus oryzae CJ-2114가 생성하는 polygalactur-onase의 최대활성을 위한 pH는 4.0, 최적온도는 4$0^{\circ}C$ 였으며, 효소활성화 에너지는 2.049㎉/㏖이었다. 정제효소의 Km값과 $V_{max}$ 값은 54.05mM, 13.9m mole/min이었고, pectin보다 polygalacturonic acid를 특이적으로 분해하였다. 이 효소는 약산성의 pH에서 안정성을 보였으며, 온도에 의한 안정성은 4$0^{\circ}C$였으며, 5$0^{\circ}C$이상에서는 급격한 효소단백질의 불활성화가 진행되었다. 금속이온중 $Na^{+}$ 이온에 의해 활성이 유지되며 C $u^{2+}$, P $b^{2+}$, $Mn^{2+}$, $Zn^{2+}$이온 등은 효소활성을 저해하였다. 효소저해제 중 maleic anhydride와 iodine등에 의해 효소활성이 저해되어 효소분자중의 histidine의 imidazole기 와 cystein의 SH기가 효소활성에 관여함이 입증되었다. 이 효소의 기질분해양상을 여지 크로마토그라피로 확인한 결과 반응초기에는 monomer, dimer, oligomer 등이 생성되었고, 시간이 경과할수록 oligomer는 사라지고 monomer, dimer만이 생성되는 것으로 보아 endo 형인 것으로 판단되었으며, 효소의 과즙청징도 측정에서는 약 18.2$\times$$10^3$unit에서 거의 청징화 되었다.

• 한국식품과학회지
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• 제38권3호
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• pp.419-426
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• 2006

전국 각지에서 채집한 토양에서 분리한 25종의 내열성 균주 중 내열성 단백질 기수분해효소 활성을 갖는 균주 strain JE 375를 선별하였다. 본 균주는 gram 양성 간균의 특징을 나타냈으며 Bergey's Manual of Systematic Bacteriology와 Biochemical Tests for Identification of Medical Bacteria에 준하여 생화학적 특성을 검토한 결과 catalase 양성, 포자형성, motility 양성, glucose 발효, mannitol 발효, xylose 산화, hemolysis ${\beta}$균임을 보아 Bacillus sp.으로 추정 되었다. Strain JE 375의 whole cell fatty acid를 gas chromatography로 분석한 결과 $C_{15:0}$ iso 26.17%, $C_{16:0}$ iso 13.01%, $C_{17:0}$ iso 30.19%로 분석되어 Bacillus 계열로 동정되었다. 16S rDNA sequence분석 결과 strain JE 375는 Bacillus caldoxylolyticus와 sequence가 97.6% 일치하는 유사성을 보였으나 부분적으로 sequence의 차이가 있고 gene bank data base상에서 16S rDNA sequence가 일치하는 균주는 검색되지 않았다. 이 같은 실험 결과에 따라 strain JE 375는 기존에 발표되지 않은 새로운 균주로 판단되어 Bacillus sp. JE 375로 명명하였다. Bacillus sp. JE 375은 tryptone 1%, yeast extract 0.5%, NaCl 1%, maltose 1%의 배지조성분과 배양 온도 $65^{\circ}C$에서 20시간 동안 배양하였을 때 최대의 단백질 분해 효소를 생산하였다. Bacillus sp. JE 375로부터 단백질 분해 효소를 acetone으로 침전시키고 DEAE-sepharose column chromatography를 통하여 효소를 정제하여 SDS-PAGE를 통해 확인한 결과 55 kDa 크기의 band를 확인할 수 있었다. 이 효소의 최적 배양 온도는 $65^{\circ}C$이었으며 배지의 최적 pH는 6.5로 나타났다. pH에 대한 안정성은 중성 부근의 pH에서 효소 활성의 안정성이 높게 나타났다. 본 효소의 반응 조건을 검토한 결과 중성 조건에서 안정하였으며, $60^{\circ}C$의 고온에서 활성을 가졌다. 효소 활성은 1 mM $CaCl_2$ 첨가에 의해 증가하였다.

• 한국미생물·생명공학회지
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• 제17권2호
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• pp.148-153
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• 1989

Cyclodextrin(CD) 생성효소(CGTase)를 생산하는 미생물을 토양으로부터 알칼리 (pH10.3) 조건하에서 1주 분리하여 동정한 결과, pH 7.5~ll.0까지 생육하는 절대 호알칼리성 Bacillus sp.이었다. 분리균주가 생산하는 효소는 전분으로부터 $\beta$-CD를 우선적으로 생성하는 효소임을 HPLC를 통해 분석하였으며, 이때의 $\alpha$-, $\beta$-, ${\gamma}$-CD의 생성비율은 1:10:1.5이었으나 분리균주의 효소생산 조건은 1% $Na_2$CO$_3$가 탄소원으로, 5% corn steep liquor를 질소원으로 하여 호기적 조건에서 37$^{\circ}C$, 48~60시간 진탕배양하였을 때 배양상징액으로 부터 최대 효소생산을 얻었다. Ethanol 침전방법을 이용하여 얻은 조효소액의 효소적 특성에 있어서 본 효소의 최적활성 pH는 6.0이었으며 pH 안정성은 pH 5~9까지 안정하였다. 최적활성 온도는 5$0^{\circ}C$이었으며 열안정은 5$0^{\circ}C$까지 안정함을 보였다. 본 효소의 활성염색 전기영동 결과, 대조균인 호알칼리성 Bacillus sp. ATCC 21783의 효소와 비교시 단백질의 성상에 있어서 차이점을 확인할 수 있었다.