• The Korean Journal of Mycology
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• v.11 no.3
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• pp.109-114
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• 1983

${\beta}-Galactosidase$ activity of Aspergillus phoenicis was studied using ONPG and lactose as substrate. It increased monotonically during the exponential growth phase and dropped rapidly at the beginning of the stationary one. It exhibited high tolerable temperature and acidic optimal pH which provides certain advantages from the industrial view point. Enzyme of ${\beta}-galactosidase$ had more subsrate affinity for ONPG than for lactose and its apparent maximum activity was also higher with the former as substrate. Activity of this enzyme depended upon the conditions of immobilization. Optimum crosslinking reaction was occurred at pH 7.2 and 0. 35 vol. % of glutaraldehyde concentration.

• Microbiology and Biotechnology Letters
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• v.11 no.3
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• pp.205-210
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• 1983

Extracellular $\beta$-galactosidase was prepared from a culture of Lactobacillus sporogenes, a spore-forming lactic acid bacterium. The enzyme functioned optimally at pH 6.8 and at 6$0^{\circ}C$ o-nitrophenyl-$\beta$-D-galactopyranoside (ONPG) in 0.05M sodium phosphate buffer. The activation energy of the enzymatic hydrolysis of ONPG was about 16,000 cal/mole below $50^{\circ}C$ and 11,300 cal/mole above the temperature. It was fairly stable over a pH range from 4.0 to 8.0 losing only less than 30% of its activity after hearting at 6$0^{\circ}C$ and pH 6.8 for 3 hours. Metal ions showed no significant effect on the enzyme activity, whereas L-cysteine exerted a slight stimulatory effect at the concentration of 10mM. The km values were 1.48mM for ONPG and 64.5mM for lactose. Hydrolysis of ONPG by the enzyme was product-inhibited by galactose (Ki=13.3mM, competitive inhibition) and by glucose(Ki= 11.4mM, uncompetitive type). The enzyme activity was also noncompetitively inhibited in the presence of lactose (Ki= 17.8mM).

• Analytical Science and Technology
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• v.5 no.4
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• pp.465-469
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• 1992

An analytical procedure is presented for the quantitative determination of lactose, glucose, and galactose in the hydrolyzate of lactose by ${\beta}$-galactosidase with high-performance liquid chromatography. An Aminex HPX-87C column at $85^{\circ}C$ and refractive index detector were used to resolve lactose, glucose, and galactose in only 12 minutes with distilled and deionized water as a mobile phase. The validity of high-performance liquid chromatography as a method for the assay of ${\beta}$-galactosidase was supported by recovery experiments and comparision of results with those by ONPG method, a spectrophotometric assay. The procedure was appropriate for determination of sugars in the enzyme reaction mixture and could by applied to analysis of ${\beta}$-galactosidase activity.

• Microbiology and Biotechnology Letters
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• v.13 no.4
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• pp.355-359
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• 1985

The purified $\beta$-galactosidase from L. sporogenes was most active at pH 7.0 and 6$0^{\circ}C$ with O-nitrophenyl-$\beta$-D-galactopyranoside (ONPG) in 0.05 M phosphate buffer. It was stable over a pH range from 5.0 to 9.0 and lost less than 10％ of its activity after heating for 30 minutes at 6$0^{\circ}C$ and pH 7.0. All the mineral ions examined in this work showed no significant activating effect, whereas L-cysteine exerted a great stimnlatory effect on the enzyme activity at the concentration of 10 mM. The Km values were 1.2 mM for ONPG and 33.3 mM for lactose. Approximately 85％ of lactose in cow's milk, in 10％ skim milk and in 5％ lactose solution was hydrolyzed after 4 hours incubation at 6$0^{\circ}C$ with 2 units of the purified $\beta$-galactosidase per $m\ell$ of the substrate solutions. The $\beta$-galactosidase from L. sporogenes, therefore, is considered to be suitable for hydrolysis of lactose in milk and other dairy products.

• Journal of Microbiology and Biotechnology
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• v.13 no.1
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• pp.134-138
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• 2003

The secondary and tertiary structures of ${\beta}$-galactosidase from L. lactis ssp. lactis 7962 were designed using Nnpredict and Sybyl version 6.3. By using site-directed mutagenesis, the mutated enzymes, Tyr-475-phe and Glu-506-Asp, were generated based on the structural modeling of L. lactis ssp. lactis 7962. The enzymes Tyr.-475-Phe and Glu-506-Asp had <$1\%$ of the activity of the native enzyme with ONPG as substrate. The $V_{max}$ values of the mutated enzymes were greatly reduced (1,800~40,000-1314) compared with the value for the native ${\beta}$-galactosidase. However, the $K_m$ values of Tyr-475-Phe and Glu-506-Asp with ONPG, PNPG, PNPF, and PNPA were not significantly different from those of the native enzyme. The results obtained support the suggestion that Tyr-475 and Glu-506 constitute very important parts of the catalytic machinery of the ${\beta}$-galactosidase.

• Korean Journal of Food Science and Technology
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• v.12 no.1
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• pp.45-52
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• 1980

The nylon (poly 6, 10) microcapsules containing ${\beta}-galactosidase$ were obtained by the interfacial polymerization of 1, 6-diaminohexane and sebacoyl chloride with ${\beta}-galactosidase$ from Escherichia coli. They were generally spherical and had a mean diameter of $80{\mu}$ with 45 % of the activity recovery. In particular, there was no transport hamper of lactose through the membrane of microcapsules. The characteristics of the microencapsulated enzyme were similar to those of soluble enzyme optimal pHs, $7.0{\sim}7.2$ for the soluble and $7.3{\sim}7.5$ for the microencapsulated ; optimal temperatures, $50^{\circ}C$ for both ; apparent $K_m,\;3.33{\times}10^{-4}(on ONPG),$ $2.86{\times}10^{-3}$ M(on lactose) for the soluble and $5.28{\times}10^{-4}$ (on ONPG), $4.25{\times}10^{-3}$ M (on lactose) for the microencapsulated ; activation energies, 8.94 for the soluble and 9.78 Kcal/mole for the microencapsulated enzyme. Using this microencapsulated ${\beta}-galactosidase$, hydrolyses of lactose and milk lactose were carried out and 80 % of 5 % lactose solution and 70 % of lactose in skim milk were hydrolyzed in 40 hr at $27^{\circ}C$. The reusability and operational stability showed that the remaining activity was 50 % of the original activity after 5 runs and 120 hr of total operating time at $27^{\circ}C$.

• Journal of environmental and Sanitary engineering
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• v.14 no.4
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• pp.180-187
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• 1999

The following is experimental result of Biochemical characteristics of Acetobacter sp. Acetobacter sp. were gram negative, short rod, non-spore-forming and motile. It reacted positively catalase, methyl red, oxidation fementation, voges-proskauer and nitrate reduction tests and hydrogen sulfide test negative and ONPG negative. Acetobacter sp. showed normal growth curve in Carr broth and there was no significant difference between isolates and type strains such as Acetobacter aceti(KCTC 1010), Acetobacter diazotrophicus(KCTC 2859). Acetobacter liquefaciens(KCTC 2804) and Acetobacter pasteurianus(KCTC 1008).

• Korean Journal of Food Science and Technology
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• v.21 no.1
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• pp.164-172
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• 1989

The formation of galactooligosaccharides by transgalactosidation reactions during hydrolysis of lactose by the ${\beta}-galactosidase$ from Streptococcus thermophilus 510 was investigated. Three oligosaccharides were detected during hydrolysis. It was found that the optimum conditions for the production of oligosaccharides was 40% lactose treated with ${\beta}-galactosidase(50\;ONPG\;units/ml)$ at $37^{\circ}C$ for 4 hours. The oligosaccharides formed accounted for 30% of the total sugars when the lactose had been 94% hydrolysed. 69% of the oligosaccharides were identified as $6-o-{\beta}-D-galactopyranosyl-D-glucose(allolactose)$ and 23% as $6-o-{\beta}-D-galactopyranosyl-D-galactose(isogalactobiose)$. The separation of galactooligosaccharides by the use of Bio-Gel P-2 gel permeation chromatography was also studied.

• Korean Journal of Microbiology
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• v.18 no.1
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• pp.7-14
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• 1980

A thermostrable ${\beta}-galactosidase$ (${\beta}-galactoside$ galactohydorlase, EC 3.2.1.23) was inducible in Bacillus coagulans by lactose and D-glactose. The enzyme was purified 87 fold, and the optimum temeprature and pH for actiivity were determined to be $60^{\circ}C$ and pH 7.5, respectively. Kinetic determinations at $55^{\circ}C$ established a Km of 3.3mM for the chromogenic substrate onitorphenyl ${\beta}-D-galactopyranoside$ (ONPG). Galactose and lactose were competitive inhibitors with Ki of 6.1mM and 4.9mM, respectively. The enzyme ws relatively thermostable. The crude enzyme was inactivated about 20% after 20 min of exposure at $60^{\circ}C$ and the purified was about 50%. Maximal enzyme activity required $Mn^{++}$, and for the thermal stabilization $Fe^{++}\;and\;Ca^{++}$ were necessary.

• Journal of Microbiology and Biotechnology
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• v.4 no.4
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• pp.343-348
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• 1994

Enterobacter sp. producing -$\beta$-galactosidase with high transgalactosylation activity was isolated from dairy wastewater. The isolate had common biochemical features to E. aerogenes and E. cloacae. Enzyme production increased as the cell mass increased with optimum enzyme activity of 0.21 Unit/mg-protein (o-nitro-phenyl-$\beta$ -D-galactoside (ONPG) as substrate) until 8 hr of culture. Whole cells permeabilized by toluene were used to produce galacto-oligosaccharide. Optimum toluene concentration, temperature and pH for -$\beta$-galactosidase activity of permeabilized whole cells were 10% (v/v), $50^{\circ}C$ and 6.0, respectively. A maximum of 38% (w/w) of galacto-oligosaccharide was obtained with lactose concentration of 20% (w/w) at $40^\{\circ}C$ and pH 6.0.