• 미생물학회지
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• 제21권2호
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• pp.51-60
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• 1983

Cultural characteristics of Strptomyces griseolus isolated from the soil were investigated. This strain was disclosed to utilize D-xylose, and D-glactose in preference order as a carbon source with the formation of glucose isomerase. The addition of sweet potato starch also proved effective promoting the total enzyme activity measured at 29% higher than the control. Corn cob, one of waste agricultural resources, was hydrolyzed in 2~3% $H_2SO_4$ solution at $100^{\circ}C$, 3~5 hours to produce a xylose syrup which gave rise to the recovery of 19.9% in a batch system and 28.2% in a repeated system. By the addition of both 2% of xylose syrup(Be'28) prepared by and us 65% of corn steep liquor (total nitrogen 1.2%), enzyme induction was maximized. The enzyme activity was stimulated by the xylose and the cell growth by the C.S.L. Also, remarkable increase of enzyme activity was noticed by the addition of protein acid hydrolysate 86.2% higher than the control. $QO_2$ of the biomass cultured in 30L capacity jarfermentor recorded low oxygen requirement of 251.2 1/hr. Maximum activity of glucose isomerase was observed noted at the 9th hour after inoculation which is 2 hours faster than the stationery was observed noted at the 9th hour after inoculation which is 2 hours faster than the stationery phase of the biomass growth. Glucose isomerase from the strain was activated by adding the $Co^{++}\;and\;Mg^{++}$ with optimum temperature of $73^{\circ}C$ and pH of 7.2. Conversion ratio of 60% glucose to frutose was 42.5% after 70 hours reaction.

• Journal of Microbiology and Biotechnology
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• 제2권2호
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• pp.78-84
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• 1992

Streptomyces sp. No.8, which produced glucose isomerase was isolated from soil samples. The isolated strain, No.8, was identified as belonging to the Genus Streptomyces. A mutant strain, SM 805, showed the greatest ability to produce glucose isomerase. It was developed from the strain, No.8, by mutagenesis induced by NTG and UV treatment. The mutant strain, SM 805, produced about 7 times more glucose isomerase than the parental strain, No.8. This enzyme catalyzed the isomerization of D-xylose, D-glucose and D-ribose. It was inactive in the absence of metal ions, but was activated by the addition of $Mg^{2+}$ or $Co^{2+}$. The optimum temperature and pH for enzyme activity were $80^\circ{C}$ and pH 8.5, respectively. The enzyme was stable in a pH range of 6.0 to 10.0, and it was highly thermostable. There was no activity loss below $80^\circ{C}$, and even above $90^\circ{C}$ about 45% of its activity was retained. The reaction equilibrium was reached when about 53% fructose was present in the reaction mixture. Whole cells containing glucose isomerase from Streptomyces sp. SM 805 were immobilized by glutaraldehyde treatment. The resultant immobilized enzyme pellets showed a relatively long stability during the isomerizing reaction. The half-life of the immobilized enzyme during the operating was 45 days in the presence of 10mM $Mg^{2+}$.

• Journal of Microbiology and Biotechnology
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• 제11권6호
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• pp.1002-1010
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• 2001

The D-xylose operon in Escherichia coli is known to be regulated by a transcriptional activator protein, XyIR, which is responsible for the expression of both xylAB and xylFGH gene clusters. The XyIR was purified to homogeneity by using the maltose binding protein fusion expression and purification systems involving two chromatography steps. The purified XyIR protein was composed of two subunits of 45 kDa, which was determined by both sodium dodecyl sulfate polyacrylamide gel electrophoresis and gel filtration. The purified XyIR was specifically bounded to the xylA promoter, regardless of adding xylose to the reaction mixture, but binding of XylR was specifically bounded to the xylA promoter, regardless of adding xylose to the reaction mixture, but binding of XylR to the xylA promoter was enhanced by adding xylose. The enhanced binding ability of XyIR in the presence of xylose was not diminished by adding glucose. The presumed XyIR binding site is located between 120 bp to 100 bp upstream the xylA initiation codon.

• Applied Biological Chemistry
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• 제26권4호
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• pp.222-230
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• 1983

본 연구에서는 저자등이 분리한 바 있는 Streptomyces griseolus가 분비한 xylose 이성화 효소(D-xylose ketol isomerase, EC 5.3.1.5, 포도당 이성화 효소)의 고정화 및 이 고정화 효소에 의한 파일롯트 규모의 이성화당 생산에의 응용을 시도하였다. 세포내 분비된 Xylose 이성화 효소를 함유한 미생물 균체를 호모 게나이저로 $500kg/cm^2$압력하에 파쇄한 결과 원 효소 역가의 98.8%가 얻어졌고, lysozyme으로 분해 했을 때는 54.7%가 얻어졌다. 이 효소의 고정화를 위한 담체로서는 Diaion HP 20, Duolite A-7, Amberlite IRA 93 및 94와 같은 포리스형 수지류가 효과적임이 밝혀졌고, Amberlite IRA 93에 대한 재생 형태는 $BO_4--$가, Diaion HP 20에 대해서는 $HCO_3-$가 효과적이었다. Amberline IRA 93에 대한 고정화 최적 조건은 pH 8.0에 $55^{\circ}C$로서 80.6%의 효소 고저오하 수율을 나타내었으며 효소 활성 반감기는 $65^{\circ}C$에서 24일 이상이었다. Amberlite IRA 93에 효소를 탈착(脫着) 실험한 결과, 담체는 재 사용이 가능 하였으며 2,3,4,5회째의 효소의 재 고정화율은 각각 98.2, 93.3, 90.7 및 87.5%를 나타내었다. 고정화 효소의 최적 반응 온도는 $60{\sim}70^{\circ}C$로 원효소의 경우에 비하여 다소 낮아지면서 폭이 넓어졌으며, 최적 pH는 $8.0{\sim}8.3$으로 알카리 쪽으로 이동하였다. 파일롯트 규모로 본 고정화 효소 충전탑(내경 30cm, 높이 85cm)에 의한 이성화당의 생산을 시도하였던바, 고정화 효소(350 IXIU/ml-R) 1리터가 30일동안에 약 293리터의 이성화당을 생산할 수 있는 것으로 나타났다.

• 한국식품영양학회지
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• 제2권1호
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• pp.1-11
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• 1989

Glucose isomerase (E.C.5.3.1.5) which reversibly catalyzes reaction between D-glucose and D-fructose was demonstrated in cell free extracts of alkalophilic Streptomyces sp. B-2 isolated from soil The optimum temperature, pH, and pH stability were 6$0^{\circ}C$, 10.5, and 7.8, respectively. The production of Gl in xylose and yeast extract was higher than that of other carbon source and nitrogen source. The Gl production was affected by Co2+ and Mg2).

• 한국미생물·생명공학회지
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• 제8권2호
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• pp.125-133
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• 1980

토양으로부터 분리한 포도당 이성화효소를 강하게 생산하는 방사균을 Bergey's manual 8판에 따라 동정한 결과 Streptomyces antibioticus 근록의 균주이었다. 본 균의 배양액으로부터 균체를 모아서 해사를 넣고 파쇄 하여 증류수로 추출하고 Mn-처리를 하여 핵단백질을 제거한 후 황산 ammonia 분획침전(0.5∼0.8포화), 수석, DEAE-cellulose column chromatography, DEAE-sephadex (A-50) column chromatography 및 sephadex G-200에 의한 gel filtration을 거쳐 비활성도로 약 380배, 회수율 25％ 정도로 분리 정제하였다.

• 한국미생물·생명공학회지
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• 제17권4호
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• pp.385-391
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• 1989

토양으로부터 분리한 호알카리성 세균 Bacillus sp. No. 1911 파쇄하여 유안투석, DEAE-cellulose column chromatography, Sephadex G-150 gel filtration으로 포도당 이성화효소를 정제하여 효소적 특성을 조사하였다. 이 효소는 Sephadex G-200 gel filtration에서 분자량은 170,000으로 측정되었고 SDS-polyacrylamide 전기영동에서 43,000의 단일 band를 얻어, 이 효소는 4개의 동일한 subunit로 구성되었음을 나타내었다. 이 효소는 pH7.5 및 $65^{\circ}C$에서 최적활성을 나타내었으며 pH7.5에서 7$0^{\circ}C$ 까지 안정하였고 Co$^{++}$ 존재하에 6$0^{\circ}C$에서 30분간 가열하였을 때 pH 6-9까지는 비교적 안정되었다.

• Molecules and Cells
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• 제28권4호
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• pp.397-401
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• 2009

Flavonoids are a group of polyphenolic compounds that have been recognized as important due to their physiological and pharmacological roles and their health benefits. Glycosylation of flavonoids has a wide range of effects on flavonoid solubility, stability, and bioavailability. We previously generated the E. coli BL21 (DE3) ${\Delta}pgi$ host by deleting the glucose-phosphate isomerase (Pgi) gene in E. coli BL21 (DE3). This host was further engineered for whole-cell biotransformation by integration of galU from E. coli K12, and expression of calS8 (UDP-glucose dehydrogenase) and calS9 (UDP-glucuronic acid decarboxylase) from Micromonospora echinospora spp. calichensis and arGt-4 (7-O-glycosyltransferase) from Arabidopsis thaliana to form E. coli (US89Gt-4), which is expected to produce glycosylated flavonoids. To test the designed system, the engineered host was fed with naringenin as a substrate, and naringenin 7-O-xyloside, a glycosylated naringenin product, was detected. Product was verified by HPLC-LC/MS and ESI-MS/MS analyses. The reconstructed host can be applied for the production of various classes of glycosylated flavonoids.