• Title/Summary/Keyword: pH. Arabinose

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Formation of D-Glucose Isomerase by Streptomyces sp. (Streptomyces sp.에 의한 포도당 이성화효소의 생성)

  • Rhee, In-Koo;Seu, Jung-Hwn
    • Microbiology and Biotechnology Letters
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    • v.8 no.3
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    • pp.173-180
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    • 1980
  • A source of D-xylose was required for the enhanced production of D-glucose isomerase of Streptomyces sp. strain K-17. D-glucose supported the luxuriant growth of the organism as well as D-xylose, but D-glucose isomerase activity was hardly detected in the D-glucose-grown cells. When the D-glucose-grown cells were incubated aerobically for a few hours in 0.5% xylose solution in 0.05 M phosphate buffer, pH 7.0, it was found that inductive formation of D-glucose isomerase occurred in the cells without multiplication. In the non-growth phase of cells the inductive formation of D-glucose isomerase occurred because a source of nitrogen for the synthesis of enzymes was obtained from turnover of protein accumulated in cells. D-ribose, L-arabinose, D-glucose, D-mannose, citrate, succinate and tartrate could not induce the formation of D-glucose isomerase, but D-xylose could induce. Inductinn of D-glucose isomerase was repressed by D-glucose and its catabolites : glycerol, succinate and citrate. Inductive formation of the enzymes in the non-growth phase was stimulated by $Ba^{2+}$, $Mg^{2+}$ and $Co^{2+}$, and inhibited by C $u^{2+}$, C $d^{2+}$, A $g^{+}$and H $g^{2+}$. The synthesis of enzymes in the induction system composed of 0.5% xylose solution was disrupted by actinomycin D, streptomycin, chloramphenicol, kanamycin, tetracycline, p-chloromercuribenzo ate, arsenate and 2, 4-dinitrophenol, but not disrupted by mitomycin C and penicillin G.icillin G.

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Enzymatic Characterization of Salmonella typhimurium Mannitol Dehydrogenase Expressed in Escherichia coli (Salmonella typhimurium에서 유래한 Mannitol Dehydrogenase 유전자의 대장균 내 발현 및 효소특성 규명)

  • Jang, Myoung-Uoon;Park, Jung-Mi;Kim, Min-Jeong;Kang, Jung-Hyun;Lee, So-Won;Kim, Tae-Jip
    • Korean Journal of Microbiology
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    • v.48 no.2
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    • pp.156-162
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    • 2012
  • A mannitol dehydrogenase (StMDH) gene was cloned from Salmonella typhimurium LT2 (KCTC 2421) and overexpressed in Escherichia coli. It has a 1,467 bp open reading frame encoding 488 amino acids with deduced molecular mass of 54 kDa, which shares approximately 36% of amino acid identity with known long-chain dehydrogenase/reductatse (LDR) family enzymes. The recombinant StMDH showed the highest activity at $30^{\circ}C$, and pH 5.0 and 10.0 for D-fructose reduction and D-mannitol oxidation, respectively. On the contrary, it has no activity on glucose, galactose, xylose, and arabinose. StMDH can catalyze the oxidative/reductive reactions between D-fructose and D-mannitol only in the presence of $NAD^+$/NADH as coenzymes. These results indicate that StMDH is a typical $NAD^+$/NADH-dependent mannitol dehydrogenase (E.C. 1.1.1.67).

Preparation and Physicochemical Properties of Soluble Dietary Fiber Extracts from Soymilk Residue at High Temperature (두유박 수용성 식이섬유의 고온 추출물 제조와 이화학적 특성구명)

  • Park, Chun-Ho;Kim, Hyun-Jung;Moon, Tae-Wha
    • Korean Journal of Food Science and Technology
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    • v.29 no.4
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    • pp.648-656
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    • 1997
  • Thermal treatment of soymilk residue was carried out at 140, 150, and $160^{\circ}C$ for possible use as a raw material for dietary fiber, and some physicochemical properties of the extracts were investigated. Soluble dietary fiber(SDF) content of the extracts prepared under optimal conditions was more than 30% suggesting the conversion of insoluble dietary fiber to SDF. The main sugar components of the extracts were glucose, galactose, and arabinose. Analysis of the moelcular weight distribution by high performance size exclusion chromatography revealed that the proportion of high molecular weight fraction decreased and that of middle-sized polymer increased as the extraction temperature increased. The viscosity of aqueous solution of the extracts decreased with an increase in extraction temperature, but showed no trend as pH changed. The solubility increased with extraction temperature showing the highest at $160^{\circ}C$. The extract at $140^{\circ}C$ had the biggest calcium-binding capacity, which correlated with the changes in viscosity.

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Bioethanol Production Using By-product of VPP (Value Prior to Pulping) (VPP (Value Prior to Pulping) 부산물을 이용한 바이오에탄올 생산)

  • Lee, Jae-Won;Kim, Hye-Yun;Jeffries, Thomas W.;Choi, In-Gyu
    • Journal of the Korean Wood Science and Technology
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    • v.38 no.6
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    • pp.561-567
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    • 2010
  • In this study, we evaluated optimal conditions for ethanol production of the spruce hydrolysate (SH) obtained from diethyl oxalate pretreatment. Fermentable sugar concentration in SH was 29.04 g/${\ell}$ except arabinose. Monosaccharides obtained from the oligomer degradation were mainly mannose (39.26 g/${\ell}$) and galactose (12.83 g/${\ell}$). Concentration of 5-HMF and furfural which are inhibitors on ethanol fermentation were 0.09 g/${\ell}$ and 0.04 g/${\ell}$ respectively. Concentration of acetic acid and total phenolic compounds in SH were 1.4 g/${\ell}$ and 2.83 g/${\ell}$. Ethanol production using hydrolysate was 11.7 g/${\ell}$ at optimal pH 6.0 after 48 h. Specific ethanol production was 0.15 (g/(${\ell}^*h$)) at pH 5.0 and 5.5. while that was 0.24 (g/(${\ell}^*h$)) at pH 6.0. Specific ethanol production has difference depend on initial pH for fermentation. Ethanol production was 14.3 g/${\ell}$ after 48 h when xylanase 20 IU was added in SH for degradation of oligomer during fermentation. It implied that ethanol production increased by 22.2% compare with control (without xylanase).

Isolation and Characterization of a Trypsin Inhibitor and a Lectin from Glycine max cv. Large Black Soybean

  • Ye, Xiu Juan;Ng, Tzi Bun
    • Food Science and Biotechnology
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    • v.18 no.5
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    • pp.1173-1179
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    • 2009
  • Trypsin inhibitors and lectins are defense proteins produced by many organisms. From Chinese 'Large Black Soybeans', a 60 kDa lectin and a 20 Da trypsin inhibitor (TI) were isolated using chromatography on Q-Sepharose, Mono Q, and Superdex 75. The TI inhibited trypsin and chymotrypsin with an $IC_{50}$ of 5.7 and $5{\mu}M$, respectively. Trypsin inhibitory activity of the TI was stable from pH 3 to 13 and from 0 to $65^{\circ}C$. Hemagglutinating activity of the lectin was stable from pH 2 to 13 and from 0 to $65^{\circ}C$. The TI was inhibited by dithiothreitol, signifying the importance of disulfide bond. The TI and the lectin inhibited HIV-1 reverse transcriptase ($IC_{50}$=44 and $26{\mu}M$), and proliferation of breast cancer cells ($IC_{50}$=42 and $13.5{\mu}M$) and hepatoma cells ($IC_{50}$=96 and $175{\mu}M$). The hemagglutinating activity of the lectin was inhibited most potently by L-arabinose. Neither the lectin nor the TI displayed antifungal activity.

Phosphate Uptake by Acinetobacter lwoffi PO8 and Accumulation (Acinetobacter lwoffi PO8에 의한 인산흡수 및 축적)

  • Yoon, Min-Ho;Ko, Jung-Youn;Choi, Woo-Young;Shin, Kong-Sik
    • Applied Biological Chemistry
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    • v.43 no.3
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    • pp.163-168
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    • 2000
  • To remove phosphate accumulated in the soil and water, Acinetobacter lwoffi PO8 possessing a high ability to accumulate phosphate was isolated from a active sludge. Bacterium was cultured in the liquid medium containing $150\;{\mu}g/mL$ of phosphate at $30^{\circ}C$ in different culture conditions to examine intracellular phosphate uptake. The initial pH in the range of $7.5{\sim}8.5$ was effective on the growth and phosphate uptake of the strain. Glycerol and arabinose used as a carbon sources showed 93 and 91% the phsphate uptake, respectively. Among the nitrogen sources, ammonium salt such as $NH_4NO_3$ and $(NH_4)_2SO_4$ was effectively utilized on the phosphate uptake compared with amino compounds. The rate of phosphate uptake of $NH_4NO_3$, and $(NH_4)_2SO_4$, was 95 and 96%, respectively The growth and Phosphate uptake ability in the strain were significantly promoted when metal ions were added in the medium; $Co^{2+}$, however, was not utilized by the strain. The capacity of phosphate uptake was enhanced to $10{\sim}20%$ when arginine, methionine, or lysine was added. Using $^{32}P$ to examine the uptake Pattern of intracellular phosphate, experiment result showed that polyphosphate was largely found in the fraction of intracellular inorganic phosphate of Acinetobacter lwoffi PO8.

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Effects of Feeding Xylose on the Growth of Broilers and Nutrient Digestibility as well as Absorption of Xylose in the Portal-drained Viscera

  • Peng, Y.L.;Guo, Y.M.;Yuan, J.M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.8
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    • pp.1123-1130
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    • 2004
  • Two experiments were conducted to examine the effects of dietary inclusion of xylose on growth performance, nutrient digestibility and xylose absorption in the portal-drained viscera of broiler chicks. In Exp. 1, ninety male 14 day-old broilers were used to study the effects of different inclusion levels (0, 5, 10, 20 and 40%) of D-xylose in the semi-purified diets on the growth and nutrient digestibility of broilers. In Exp. 2, One hundred and eight male broilers, fed by precision feeding at 22 day-old, were used to investigate the absorption and transportation of dietary xylose in the portal-drained viscera of broiler. The results of Exp. 1 indicated that the growth of broilers was gradually decreased as the xylose level increased (p<0.01). With the xylose supplementation increased, the moisture in broiler excreta was gradually elevated (p<0.01), AME and the digestibilities of crude protein and ether extract were significantly reduced and the digestibilities of xylose and arabinose were also decreased (p<0.01). The results of Exp. 2 showed that the concentrations of ribose, xylose and galactose in serum were significantly influenced by different dietary levels of xylose (p<0.01), but there's no apparent difference among rhamnose, glucose and arabinose (p>0.05). The xylose concentration in serum was highest in Vena Cava, middle in Portal Vein and lowest in Ulnar Vein within 6 h after precision feeding. And then the xylose concentration in Portal Vein and Ulnar Vein were higher than that of Vena Cava. The concentration of ribose, xylose and galactose in serum were also significantly changed with time prolongation (p<0.01). The concentration of xylose in serum was highest in the 40% xylose treatment, middle in the 20% xylose group and lowest in the control group. The glycogen contents in liver and muscle were linearly decreased as the level of xylose increased (p<0.01). The higher the dietary level of xylose was, the lower digestibility of dietary xylose was (p<0.10). 40% xylose markedly decreased the digestibility of dietary glucose (p<0.01). In conclusion, high levels of xylose in the diets inhibited the growth and nutrient digestibility of broiler. The outputs of xylose from the hydrolyzation of wheat-based diet by xylanase should have no adverse effects on broiler performance.

Purification of Oat ${\beta}-Glucan$ by ${\alpha}-Amlyase$ Treatment and Characterization of Its Physicochemical Properties (귀리 ${\beta}-glucan$${\alpha}-amlyase$를 이용한 정제와 이화학적 특성)

  • Park, Hee-Joeng;Kang, Tae-Su;Lee, Hee-Bong;Kim, Kwang-Yup;Jang, Keum-Il;Noh, Young-Hee;Jeong, Heon-Sang
    • Korean Journal of Food Science and Technology
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    • v.37 no.5
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    • pp.776-782
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    • 2005
  • The effects of purification using ${\alpha}-amlyase$ (Termamyl 120L) on physicochemical properties of ${\beta}-glucan$ from oat bran were studied. Four fractions were selected as fraction A ($55^{\circ}C$, 15%, pH 6), fraction B ($45^{\circ}C$, 15%, pH 6), fraction C ($50^{\circ}C$, 0%, pH 7), and fraction D ($50^{\circ}C$, 10%, pH 5) from the result of physiological test, and three consecutive subfractions were obtained by repeated ${\alpha}-amlyase$ treatments on the each fractions. The contents of ${\beta}-glucan$, protein, and ash after purification were in 81.4-88.2%, 4.1-6.3% and 2.6-6.2%, respectively. The apparent viscosities of purified ${\beta}-glucan$ aqueous solutions were similar to those of hydroxy methyl cellulose. Glucose was a major monosaccharide of ${\beta}-glucan$ extracts, and xylose and arabinose were also detected as minor constituents on TLC. The average molecular weight ranged $2.0{\times}10^6-5.1{\times}10^6$ and was decreased after purification. From the result of the differential scanning calorimetry, the melting point ranged $130-140^{\circ}C$ with purification step and thermal transition enthalpy was increased. The ratio of ${\beta}-(1{\rightarrow}3)\;to\;{\beta}-(1{\rightarrow}4) $ linkages were 1:2.22-1:2.52, and increased up to 1:5.50 after purification.

Purification and characterzation of the $\alpha$-L-Arabinofuranosidase from Escherichia coli Cells Harboring the Recombinant Plasmid pKMG11 (재조합 균주 Escherichia coli가 생산하는 Bacillus stearothermophilus $\alpha$-L-Arabinofuranosidase의 정제 및 특성)

  • 엄수정;조쌍구;최용진
    • Microbiology and Biotechnology Letters
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    • v.23 no.4
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    • pp.446-453
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    • 1995
  • $\alpha $-Arabinofuranosidase was produced by E. coli HB101 haboring the recombinant plasmid pKMG11 which contained the arfI gene of Bacillus stearothermophilus. The maximum production of the enzyme was observed when E. coli HB101 cells were grown at 37$\circ$C for 20 hours in the medium containing 0.5% arabinose, 1.0% tryptone, 0.5% yeast extract, and 1% NaCl. The $\ALPHA $-arabinofuranosidase produced was purified to homogeneity using a combination of 20-50% ammonium sulfate precipitation, DEAE-Sepharose CL-6B ion exchange column chromatography and Sepharose 6B-100 gel filtration. The purified enzyme was most active at 55$\circ$C and pH 6.5. The K$_{m}$ and V$_{max}$ values of the enzyme on $\rho $-nitrophenyl-$\alpha $-arabinofuranoside was determined to be 2.99 mM and 0.43 $\mu $mole/min (319.74 $\mu $mole/min/mg), respectively. The pI value was 4.5. The molecular weight of the native protein was estimated to be 289 kDa. The SDS-polyacrylamide gel clectrophoresis analysis suggested that the functional protein was a trimer of the 108 kDa identical subunits. The N-terminal amino acid sequence of the a-arabinofuranosidase was identified as X-Ser-Thr-Ala-Pro-Arg( \ulcorner )-Ala-Thr-Met-Val-Ile-Asp-X-Ala-Phe.

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Studies on the Production of Alcohol from Woods (목재(木材)를 이용(利用)한 Alcohol 생산(生産)에 관(關)한 연구(硏究))

  • Cheong, Jin Cheol
    • Journal of Korean Society of Forest Science
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    • v.59 no.1
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    • pp.67-91
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    • 1983
  • In order to examine the alcohol production from softwoods (Pinus densiflora Sieb. et Zucc., Pinus rigida Miller, Larix leptolepis Gordon) and hardwoods (Alnus japonica Steud., Castanea crenata Sieb. et Zucc. Populus euramericana CV 214), chemical compositions were analyzed and conditions of acid hydrolysis with wood meals were established. Also strains which could remarkably decompose the cellulose were identified, and conditions of cellulase production of strains, characteristics of cellulase, and alcohol fermentation were examined. The results were summarized as follows. 1) In acid hydrolysis of wood, the high yield of reducing sugars was shown from 1.0% to 2.0% of hydrochloric acid and 2.0% of sulfuric acid. The highest yield was produced 23.4% at wood meals of Alnus japonica treated with 1.0% of hydrochloric acid. 2) The effect of raising the hydrolysis was good at $1.5kg/cm^2$, 30 times (acid/wood meal), and 45 min in treating hydrochloric acid and 30 min in treating sulfuric acid. 3) The pretreatments with concentrated sulfuric acid were more effective concentration ranged from 50% to 60% than that with hydrochloric acid and its concentration ranged from 50% to 60%. 4) The quantative analysis of sugar composition of acid hydrolysates revealed that glucose and arabinose were assayed 137.78mg and 68.24mg with Pinus densiflora, and 102.22mg and 65.89mg with Alnus janonica, respectively. Also xylose and galactose were derived. 5) The two strains of yeast which showed remarkably high alcohol productivity were Saccharomyces cerevisiae JAFM 101 and Sacch. cerevisiae var. ellipsoldeus JAFM 125. 6) The production of alcohol and the growth of yeasts were effective with the neutralization of acid hydrolysates by $CaCO_3$ and NaOH. Production of alcohol was excellent in being fermented between pH 4.5-5.5 at $30^{\circ}C$ and growth of yeasts between pH 5.0-6.0 at $24^{\circ}C$. 7) The production of alcohol was effective with the addition of 0.02% $(NH_2)_2CO$ and $(NH_4)_2SO_4$, 0.1% $KH_2PO_4$, 0.05% $MgSO_4$, 0.025% $CaCl_2$, 0.02% $MnCl_2$. Growth of yeasts was effective with 0.04-0.06% $(NH_2)_2CO$ and $(NH_4)_2SO_4$, 0.2% $K_2HPO_4$ and $K_3PO_4$, 0.05% $MgSO_4$, 0.025% $CaCl_2$, and 0.002% NaCl. 8) Among various vitamins, the production of alcohol was effective with the addition to pyridoxine and riboflavin, and the growth of yeasts with the addition to thiamin, Ca-pantothenate, and biotin. The production of aocohol was increased in 0.1% concentration of tannin and furfural, but mas decreased in above concentration. 9) In 100ml of fermented solution, alcohol and yeast were produced 2.201-2.275ml and 84-114mg for wood meals of Pinus densiflora, and 2.075-2.125ml and 104-128mg for that of Alnus japonica. Residual sugars were 0.55-0.60g and 0.60-0.65g for wood meals of Pinus densiflora and Alnus japonica, respectively, and pH varied from 3.3 to 3.6. 10) A strain of Trichoderma viride JJK. 107 was selected and identified as its having the highest activity of decomposing cellulose. 11) The highest cellulase production was good when CMCase incubated for 5 days at pH 6.0, $30^{\circ}C$ and xylanase at pH 5.0, $35^{\circ}C$. The optimum conditions of cellulase activity were proper in case of CMCase at pH 4.5, $50^{\circ}C$ and xylanase at pH 4.5, $40^{\circ}C$. 12) In fermentation with enzymatic hydrolysates, the peracetic acid treatment for delignification showed the best yields of alcohol and its ratio was effective with the addition of about 10 times. 13) The production of alcohol was excellent when wood meals and Koji of wheat bran was mixed with 10 to 8 and the 10g of wood meals of Pinus densiflora produced 2.01-2.14ml of alcohol and Alnus japonica 2.11-2.20ml.

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