• Korean Journal of Microbiology
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• v.15 no.1
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• pp.9-19
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• 1977

Xylone isomerase (D-xylose ketol-isomerase, EC 5,3,1,5) have been demonstrated in the cell-free extracts of Stroptomuces canus and Streptomuces malachiticus grown in the presence of xylose. Xylose, glucose and ribose served as substrates for the enzymes of the two strains with respective $K_m$ values of 22, 130, 290 mM (S. canus) and 7,83,637 mM(S.malachiticus), and $V_max$ values of 1,000, 0.087, $\0.0222{\mu}moles/min/mg$ protein (S. canus) and 0.312, 0.083, 0.500.$\mu$moles/min/mg protein (S. malachiticus). L-Rhammose was also isomerized by the crude enzyme solutions of the two strains. The maximal activities of the two xylose-isomerases were observed at pH 7.5 and $75^{\circ}C$. The xylose isomerase activities of the two strains were activated two-three times by $Mg^{++}\;and\;Co^{++}$ as that of control, partially activated by $Ba^{++}$ and inhibited by $Ni^{++},\;Ca^{++}\;and\;Zn^{++}\$. Particulary, the addtion of $Mn^{++}$ stimulated xylose-isomerizing activities, but inhibited glucose-isomerizing activities in both strains. However, $Cu^{++}$ inhibited xylose-isomerizing activities, while stimulated glucose-isomerizing activities of the enzymes.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.1
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• pp.27-31
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• 2000

Characteristics of ethanol production by a xylose-fermenting yeast, Pichia stipitis Y-7124, were studied. The sugar consumption rate and specific growth rate were higher in the glucose-containing medium than in the xylose-containing medium. Specific activities of xylose reductase and xylitol dehydrogenase were higher in the medium with xylose than glucose, suggesting their induction by xylose. Maximum specific growth rate and ethanol yield were achieved at 30 g xylose/L concentration without formation of by-products such as xylitol and acetic acid whereas a maximum ethanol concentration was obtained at 130 g/L xylose. Adding a respiratory inhibitor, rotenone, increased a maximum ethanol concentration by $10\%$ compared with the control experiment. In order to evaluate the pattern of ethanol inhibition on specific growth rate, a kinetic model based on Luong's equations was applied. The relationship between ethanol concentration and specific growth rate was hyperbolic for glucose and parabolic for xylose. A maximum ethanol concentration at which cells did not grow was 33.6 g/L for glucose and 44.7 g/L for xylose.

• Journal of Microbiology and Biotechnology
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• v.18 no.5
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• pp.837-844
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• 2008

Glucose (xylose) isomerases from Streptomyces sp. have been used for the production of high fructose corn syrup for industrial purposes. An 11-kb DNA fragment containing the xyl gene cluster was isolated from Streptomyces lividans TK24 and its nucleotide sequences were analyzed. It was found that the xyl gene cluster contained a putative transcriptional repressor (xylR), xylulokinase (xylB), and xylose isomerase (xylA) genes. The transcriptional directions of the xylB and xylA genes were divergent, which is consistent to those found in other streptomycetes. A gene encoding XylR was located downstream of the xylB gene in the same direction, and its mutant strain produced xylose isomerase regardless of xylose in the media. The enzyme expression level in the mutant was 4.6 times higher than that in the parent strain under xylose-induced condition. Even in the absence of xylose, the mutant strain produce over 60% of enzyme compared with the xylose-induced condition. Gel mobility shift assay showed that XylR was able to bind to the putative xyl promoter, and its binding was inhibited by the addition of xylose in vitro. This result suggested that XylR acts as a repressor in the S. lividans xylose operon.

• 한국신재생에너지학회:학술대회논문집
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• 2005.11a
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• pp.286-298
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• 2005

농산 폐자원으로부터 식품 및 의약품 소재로 쓰이는 xylose, arabinose, cellulose를 생산하였다. 농산 폐자원은 우리나라 실정에 적합한 볏짚과 옥수수 껍질을 선택하였으며, 공정수율은 xylose와 arabinose는 약 15$\sim$20%(w/w), cellulose 는 20%(w/w)로 나타났다. 폐자원을 활용하는 개발된 공정중에는 미생물 유전자 재조합 기술을 응용하여 고역가의 효소생산 system을 개발하여, 생산된 효소를 가수분해 공정 과정에 투입하여 경제성이 높고 친환경적인 기술로 확립하였다. 재조합 미생물과 xylose, arabinose 정제공정은 신뢰 높은 재연성을 나타냈으며 xylose 제조법과 xylitol 발효법은 package 형태로 기술 이전을 준비하고 있다.

• Microbiology and Biotechnology Letters
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• v.40 no.2
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• pp.163-167
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• 2012

Since D-xylose is not fermentable in Saccharomyces cerevisiae, its conversion to D-xylulose is required for its application in biotechnological industries using S. cerevisiae. In order to convert D-xylose to D-xylulose by way of an enzyme immobilized system, D-xylose isomerase (XI) of Escherichia coli was fused with 10-arginine tag (R10) at its C-terminus for the simple purification and immobilization process using a cation exchanger. The fusion protein XIR10 was overexpressed in recombinant E. coli and purified to a high purity by a single step of cation exchange chromatography. The purified XIR10 was immobilized to a cation exchanger via the electrostatic interaction with the C-terminal 10-arginine tag. Both the free and immobilized XIR10 exhibited similar XI activities at various pH values and temperatures, indicating that the immobilization to the cation exchanger has a small effect on the enzymatic function of XIR10. Under optimized conditions for the immobilized XIR10, D-xylose was isomerized to D-xylulose with a conversion yield of 25%. Therefore, the results of this study clearly demonstrate that the electrostatic immobilization of XIR10 via the interaction between the 10-arginine tag and a cation exchanger is an applicable form of the conversion of D-xylose to D-xylulose.

• Journal of Life Science
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• v.14 no.4
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• pp.636-643
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• 2004

D-Xylose isomerase produced by Lactococcus sp. JK-8, isolated from kimchi, was purified 17-fold of homogeneity, and its physicochemical properties were determined. Although the N-terminal amino acid sequence of D-xylose isomerase was analysed to Ala-Tyr-Phe-Asn-Asp-Ile-Ala-Pro-Ile-Lys, it was not similar to that of Lactobacillus enzyme. The molecular weight of the purified enzyme was estimated to be 180 kDa by gel filtration, 45 kDa by SDS-PAGE and the enzyme was homotetramer. The optimum pH of the enzyme was around 7 and stable between pH 6 and 8. The optimum reaction temperature was 7$0^{\circ}C$ and stable up to 7$0^{\circ}C$ in the presence of 1 mM $Mn^{2+}$. Like other D-xylose isomerases, this enzyme required divalent cation, such as $Mg^{2+}$, $Co^{2+}$, or $Mn^{2+}$ for the activity and thermostability. $Mn^{2+}$was the best activator. Substrate specificity studies showed that this enzyme was highly active on D-xylose. The enzyme had an isoelectric point of 4.8, and fm values for D-xylose was 5.9 mM.

• Journal of Microbiology and Biotechnology
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• v.11 no.6
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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.

• Microbiology and Biotechnology Letters
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• v.15 no.5
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• pp.340-345
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• 1987

Microorganisms capable of utilizing D-xylose as a sole carbon and energy source were isolated to ferment D-xylose directly to ethanol. Among them, the strain, which showed the best ability to pro-duce ethanol, was selected and was identified as Fusarium sp. The optimal conditions for the pro-duction of ethanol were 8.0 of initial pH, 33$^{\circ}C$ of temperature, and 2% of substrate concentration. Under this optimal condition, the following results were obtained : maximum ethanol concentration, 7.0g/$\ell$; ethanol yield, 0.35g of ethanol per g of D-xylose (68.6% of theoretical); biomass yield, 0.27g of dry biomass per g of D-xylose.

• Korean Journal of Microbiology
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• v.28 no.4
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• pp.345-350
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• 1990

In order to ferment D-xylose directly to ethanol, Yeasts capable of utilizing D-xylose as a sole carbon source and energy source were isolated from soil, sawdust and rotten woods. Among them, the yeast strain, which showed the best ability to produce ethanol, was identified as Candida sp. L-16 isolated from rotten woods. The optimal conditions for production of ethanol were 60rpm of agitation speed, 28j.deg.C of temperature, 4.5 of initial pH and 5% of D-xylose concentration. Ethanol production was reached to maximum state for 4 days culture. Under these optimal conditions, the maximum ethanol concentration and theoretical ethanol yield were 2.4%(v/v) and 74.4% of theoretical value, respectively.

• Microbiology and Biotechnology Letters
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• v.11 no.3
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• pp.193-199
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• 1983

Systematic bioconversion process for the production of xylose from agricultural wastes such as barley straw and corn cobs was studied. After the pretreatment in 1 % NaOH solution for 24 hours at 3$0^{\circ}C$, enzymatic hydrolysis of barley straw for 48 hours at 3$0^{\circ}C$ resulted in the liberation of 15.8% of reducing sugar which is equivalent to 87% of total D-xylose content. Among various agricultural wastes, corn cob as well as barley straw was demonstrated to be potent sources for the production of D-xylose by the process of enzymatic conversion.