• 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.

• Journal of Microbiology and Biotechnology
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• v.2 no.1
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• pp.14-20
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• 1992

Some microorganisms isolated from soil, including some bacteria and fungi, were found to secrete an extracellular soymilk-clotting enzyme. Among them, an isolated fungus showed the highest soymilk-clotting activity and the strain was assigned to genus Penicillium based on its cultural and morphological characteristics, and designated as Penicillium sp. L-151K. Soymilk-clotting enzymes A and B produced by Penicillium sp. L-151K were purified by ammonium sulfate precipitation and chromatographies on Sephadex G-25, CM-Sephadex, Sephadex G-100 and phenyl-Toyopearl gel. The two purified enzymes A and B were found to be homogeneous by polyacrylamide gel electrophoresis at pH 9.5. The molecular weights of enzyme A and B were 24, 000 and 40, 000, respectively, by gel filtration on Sephadex G-100. Enzymes A and B coagulated soymilk optimally at $60^\circ{C}$ and were stable up to $50^\circ{C}$. Both enzymes were most active at pH 5.8 for soymilk coagulation, and were stable with approximately 80% of original activity from pH 3.0 to 5.0. Each enzyme was an acidic protease with an optimum pH of 3.0 for casein digestion. The soymilk-clotting efficiency of these enzymes was improved with $CaCl_2\;or\;MgCl_2$ when making soymilk-curd.

• Journal of Microbiology and Biotechnology
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• v.20 no.10
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• pp.1415-1423
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• 2010

An extracellular xylanase was purified to homogeneity by sequential chromatography of Fomitopsis pinicola culture supernatants on a DEAE-Sepharose column, a gel filtration column, and then on a MonoQ column with fast protein liquid chromatography. The relative molecular mass of the F. pinicola xylanase was determined to be 58 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis and by size-exclusion chromatography, indicating that the enzyme is a monomer. The hydrolytic activity of the xylanase had a pH optimum of 4.5 and a temperature optimum of $70^{\circ}C$. The enzyme showed a $t_{1/2}$ value of 33 h at $70^{\circ}C$ and catalytic efficiency ($k_{cat}=77.4\;s^{-1}$, $k_{cat}/K_m$=22.7 mg/ml/s) for oatspelt xylan. Its internal amino acid sequences showed a significant homology with hydrolases from glycoside hydrolase (GH) family 10, indicating that the F. pinicola xylanase is a member of GH family 10.

• Fisheries and Aquatic Sciences
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• v.15 no.1
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• pp.9-14
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• 2012

The objectives of this research were to improve the film-forming properties of Channel Catfish Ictalurus punctatus skin gelatin (CSG) by cross-linking with transglutaminase (TG), determine and optimize the TG reaction time, and characterize the mechanical and barrier properties of CSG edible film. Cross-linking of CSG was performed by TG for 0, 10, 20, 30, and 40 min at $50^{\circ}C$, and the reaction was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The color and mechanical and barrier properties of edible films fabricated with CSG cross-linked with TG were characterized. Gelatin yields from the extraction ranged from 18.2% to 23.3%. SDS-PAGE exhibited dark bands at 120 and 250 kDa, indicating successful TG-mediated cross-linking. The color of CSG film was not affected by TG cross-linking. The tensile strength of CSG films cross-linked with TG decreased from 42.59 to 21.73 MPa and the percent elongation increased from 42.92% to 76.96% as reaction times increased from 0 to 40 min. There was no significant difference in water vapor permeability of CSG films.

• Journal of the Korean Society of Food Science and Nutrition
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• v.9 no.1
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• pp.59-73
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• 1980

A detailed procedure was described for the isolation of cratine kinase (ATP-Creatine phosphotransferase, E. C. 2. 7. 3. 2.) from the muscle of the snake Bungarus fasciatus. The original isolation procedure of Kuby et al. for the rabbit muscle enzyme has been modified and extended to include a chromatographic step. The properties of the enzyme have been investigated and kinetic constants for the reverse reactions determined as the followings: 1) A molecular weight of the enzyme was determined by gel filteration on Sephadex G-100 and by electrophoresis on SDS-polyacrylamide was 86,000. 2) Two reactive sulphydryl groups were detected with dithiobis nitrobenzoic acid (DTNB). 3) The nucleotide substrate specificity in the reverse reaction was determined as ADP*2'-dADP＞GDP＞XDP＞UDP with magnesium as the activating metal ion. 4) The order of the metal specificity in the reverse reaction Mg>Mn>$Ca{\sim}Co$ was determined with ADP as substrate. 5) A detailed kinetic analysis was carried out in the reverse direction with $MaADP^-$ as the nucleotide substrate. Initial velocity and product inhibition studies($MaADP^{2-}$ competitive with respect to MgADP- and noncompetitive with respect to $N-phosphorycreatine^{2-}$ ; Creatine competitive with respect to $N-phosphorycreatine^{2-}$ and noncompetitive with respect to Ma $ADP^-)$ indicated that the reaction obeyed a sequential mechanism of the rapid equilibrium random type.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.1
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• pp.38-45
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• 1998

To extract insoluble proteins and to improve funtional properties of abolished proteins, an phytase producing Aspergillus sp. SM-15 was isolated from soil. The enzyme was purified and its enzymological characteristics were investigated. Phytase production reached to maximum when the wheat bran medium containing 1% mannose, 1% yeast extract, 1% (NH4)2HPO4 and 0.2% calcium chloride was cultured for 4 days. Phytase was purified 17.1 fold and specific activity was 244.32unit/mg by a sequencial process of ammonium sulfate fraction, ion exchange chromatography and gel filtrations Pruified enzyme was confirmed as a single band by the polyacrylamide gel electro-phoresis. The molecular weight of phytase was estimated to be 46,000. The optimum pH and temperature for the phytase activity were 5.5 and 5$0^{\circ}C$. The enzyme is stable in pH 4.5~5.5, 6$0^{\circ}C$. The activity of purified enzyme was inhibited by Hg2+ whereas activited by Pb2+ and Fe2+. The activity of phytase was inhibited by the treatment with iodine. The result indicate the possible involvement of histidine at active site. Km and Vmax of the puridied phytase were 37.037mM/L and 159.87umol/min, respectively.

• Korean Journal of Air-Conditioning and Refrigeration Engineering
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• v.8 no.3
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• pp.330-337
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• 1996

The effects of pump and temperature on drag reducing characteristics were investigated with a polymer(PAAM : Polyacrylamide) and three kinds of surfactants(CTAC, STAC, Habon-G) in fully developed turbulent pipe flows with various experimental parameters such as additive concentration(30~500ppm), pipe diameter(4.65mm, 10.85mm), Reynolds number($4{\times}10^4{\sim}10^5$) and working fluid temperature($20{\sim}80^{\circ}C$). The pump effect on PAAM was severe such that the drag reduction rates obtained with pump were decreased upto 30% as compared with those obtained with compressed air in 4.65mm test section. The temperature effect on PAAM was noticeably considerable, that is, the higher temperaute, the less drag reduction rate. On the other hand, no significant pump effect on the surfactants was observed. The drag reducing effectiveness of CTAC was totally lost in the temperature ragne of 60 to $80^{\circ}C$, whereas STAC and Habon-G kept their distinct drag reducing capability at a temperature of $80^{\circ}C$. This study clearly elucidated that for DHC application of drag reducing additives, the pump and temperature effects as well as additive concentration and pipe diameter should be carefully taken into consideration.

• YAKHAK HOEJI
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• v.33 no.6
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• pp.339-344
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• 1989

The alkaline protease produced by Sarcodon aspratus(Berk) S. Ito. was purified from its fruit bodies. The enzyme was purified by using ammonium sulfate fractionation, tris-acryl CM-cellulose column chromtography and chromatofocusing. The protease migrated as one major band with a molecular weight of about 29,000 dalton on sodium dodecylsulfate-polyacrylamide gel electrophoresis. The amino acid sequence of the N-terminal residues(21) of the enzyme was determined by automated sequence analysis. The sequence was Val-Thr-Thr-Lys-Gln-Thr-Asn-Ala-Pro-Trp-Gly-Leu-Gly-Asn-Ile-Ser-Thr-Thr-Asn-Lys-Leu. Comparison of this sequence with the N-terminal sequence of the p-roteinase K from Tritirachium album showed high similarity, i. e. 57.8% identical residues. The protease displayed a relatively high stability in sodium dodecyl sulfate.

• Journal of Environmental Health Sciences
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• v.18 no.1
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• pp.84-94
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• 1992

In an attempt to develop the immobilized biocatalysts based on immobilized Saccharomyces cerevisiae, immobilized yeast was investigated with respect to the conditions affected to ethanol productivities. Saccharomyces cerevisiae was immobilized in the form of the beads by magnetic-calcium alginate, non magnetic-calcium alginate and acrylamide polymerization. Magnetic immobilized yeast, nonmagnetic immobilized yeast and polyacrylamide immobilized yeast were compared in respect of their pH stability, thermostability, heat tolerance, the relation between the concetration of native yeast and retained activity of immobilized yeast, the activity depending on bead size of immobilized yeast, and the effects of magnesium and cobalt on the activities. The more small bead had retained the higher activity for the three kinds of immobilized yeast. In case of 1.0mm diameter of beads, the retained activity was 40~50% for the all groups. The pH stability profile for the immobilized yeast showed a broad range of optimun activity while the native yeast gave a sharp pick for its optimun pH value. The thermostability was at the range of 25~55$^{\circ}$C for the immobilized yeast groups. It was investigated that the influent magnesium and cobalt concentration, and the relative activity have an influent on heat tolerance at steady state. Both protein content released from immobilized yeast and activity of immobilized yeast were changed after activation of immobilized yeast cell.

• Journal of Environmental Health Sciences
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• v.30 no.3
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• pp.253-258
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• 2004

Synthetic organic polyelectrolytes can be used to condition sludges to enhance their dewaterability. Intermittent aerobic digestion is an useful digestion technology and has many advantages like neutral pH, low installation cost and easiness to operation. The objectives of this study were to investigate the dewaterability of intemittent aerobic digestion sludge and to find the relationship between dewaterability and particle size distribution change under the conditioning of intermittent aerobic digestion sludge by cationic polyelectrolyte. Digested sludge from intermittent aerobic digestion was used and cationic polyacrylamide polymer was added as a conditioner. CST(capillary suction time), TTF(time-to-filtration) were tested as a dewaterability index and the number of particle distribution was analyzed using particle size analyzer. The results indicate that cationic polyelectrolytes is useful to enhance dewaterability of intermittent aerobic digestion sludge. Mean particle diameter was increased as polymer dosage increased and its value was reached up to 100 mm on the condition of optimal cationic polymer dosage. CST and TTF are well correlated with mean particle diameter when the weighting order is 1.7. By the optimal conditioning with cationic polymer, particles in the filtrate are also reduced significantly and this means that conditioning is helpful to main stream by reducing SS loading from return flow.