• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 2005.05a
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• pp.322-325
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• 2005

본 연구는 ${\beta}-galactosidase$에 의한 isoflavone 배당체의 비배당화 연구를 실용화하기 위하여 바람직하지 않은 관능적 요소와 화학적 반응을 제어하기 위하여 두 물질을 미세캡슐화 하고 그것의 in vitro 안정성을 연구하여 기능성식품개발에 응용토록 하는데 목적을 두었다. 실험 결과, 미세캡슐화 수율이 매우 높았으며, 인공위액 상태에서의 안정성은 높은 반면에, 인공 소장액 상태에서의 안정성이 매우 낮았다. 또한 인공소장액에서 유리된 ${\beta}-galactosidase$에 의한 유리 isoflavone의 비배당화율이 약 75% 정도로 관찰되었다. 결과적으로 판단해 볼 때, isoflavone과 ${\beta}-galactosidase$의 미세캡슐화를 통한 소화기 내에서의 비배당화와 흡수율을 극대화하는데 매우 긍정적인 가능성을 보였다.

• 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 Aquaculture
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• v.9 no.4
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• pp.445-452
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• 1996

Endogeneous activities of ${\beta}-galactosidase$-like enzyme in various tissues from several finfishes and shellfishes were examined by histochemical analysis based on X-gal staining and by fluorimetric measurement using 4-methylumbelliferyl-${\beta}$-D-galactoside (4-MUG). Species used in this study were 3 freshwater fishes, mud loach (Misgurnus mizolepis), common carp (Cyprinus carpio) and tilapia (Oreochromis niloticus) ; 3 marine fishes, olive flounder (Paralichthys olivaceus), stone flounder (Kareius bicoloratus) and marbled sole (Limanda yokohamae) ; and 4 shellfishes, abalone (Haliotis discus hannai), Pacific oyster (Crassoskra gigas), pearl oyster (Pinctada fucata martensii) and ark shell (Anadara broughtonii). The activities of ${\beta}-galactosidase$-like enzyme in all finfishes examined were significantly different among species, with the wide variations between tissues in a species. In general, the tissues such as kidney, intestine and liver were ones which showed the significantly higher values in 4-MUG fluorimetry and deeper staining patterns in X-gal analysis compared to other tissues. On the other hand, serum and muscle revealed the significantly lower activities than others did, regardless of species. Shellfishes were also found to have endogenous activities of ${\beta}-galactosidase$-like enzyme which were significantly varied depending on both species and organs in a species. Hepatopancreas from all shellfishes examined showed the deepest pattern in X-gal staining and also the highest value in 4-MUG analysis, while activities of ${\beta}-galactosidase$-like enzyme in adductor muscles and mantle muscles from all shellfish species in this study except Pacific oyster were negligible : Pacific oyster had the significant activity of this enzyme in muscle tissues. Putative endogenous lacZ fragment was amplified from both finfishes and shellfishes by polymerase chain reaction (PCR). The molecular size of PCR products was about 510 bp, and there was no difference in size among species examined.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.758-761
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• 2001

In this study, the nar promoter of E. coli was characterized to see whether the nar promoter cloned onto pBBR122 can be used as an expression promoter of gram negative microbes. For this purpose, a plasmid with lacZ gene expressing ${\beta}-galactosidase$ instead of the structural genes of nar operon in a gram negative host strain(Agrobacterium.tumefaciens) was used to simplify an assay of induction of the nar promoter. The following effects were investigated to find optimal conditions: methods of inducing the nar promoter, optimal nitrate concentration, maximally inducing the nar promoter, the amount of expressed ${\beta}-galactosidase$ and induction ratio(specific ${\beta}-galactosidase$ activity after maximal induction/specific ${\beta}-galactosidase$ activity before induction). The following results were obtained from the experiments: the growth of Agrobacterium with E.coli nar promoter was not much affected by nitrate concentration in the shake-flask; induction of nar promoter was optimal when Agrobacterium was grown in the presence of 1% nitrate ion at the beginning of culture and when overnight culture was completely grown in the shake-flask before being transferred to other shake-flask; the amount of ${\beta}-galactosidase$ per cell and per medium volume was maximal when Agrobacterium was grown under aerobic condition to $OD_{600}$ of 1.7; then the nar promoter was induced under microaerobic and anaerobic condition made by lowering dissolved oxygen level(DO). After 2-3h of induction in the YEP medium selected as a main culture medium, the specific ${\beta}-galactosidase$ activity became about 17,000 Miller units in the fermentor cluture.

• Applied Biological Chemistry
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• v.38 no.6
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• pp.502-506
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• 1995

A Bacillus strain which produces ${\beta}-galactosidase$ with high transgalactosylation activity, was isolated from soil and tentatively designated as Bacillus sp. A1. When ${\beta}-galactosidase$ from Bacillus sp. A1 reacted with 40% (w/w) lactose, transgalactosylation ratio reached up to 90% at the 70% conversion of the initial lactose. The biosynthesis of the enzyme in Bacillus sp. A1 required lactose as an inducer and was repressed by glucose. Observing that the addition of amino acids to culture medium resulted in enhancing, to a significant extent, both the growth and the enzyme production of the strain, yeast extract and commercially available hydrolysates of protein were examined for the suitability as amino acid source. As it turned out, SMP, an enzymatic hydrolysis product of soybean protein from Fuji Oil Co.(Japan), was the most suitable for optimization of the culture medium. When Bacillus sp. A1 was cultured in the presence of 0.5% SMP and 2% lactose, the enzyme activity increased up to $1.8\;U/m{\ell}-broth$.

• Microbiology and Biotechnology Letters
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• v.37 no.3
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• pp.204-212
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• 2009

A strain producing ${\alpha}$-galactosidase and ${\beta}$-glucosidase was isolated from Kimchi. The isolated strain was identified as Weissella cibaria by 16S rDNA analysis and designated as Weissella cibaria K-M1-4. The enzyme activity of ${\alpha}$-galactosidase and ${\beta}$-glucosidase reached the maximum in the soy medium at $37^{\circ}C$ for 24 hr. The enzymes were purified by ethanol fractionation, DEAE sepharose fast flow, and sephacryl S-100HR column chromatography. ${\alpha}$-Galactosidase specific activity was shown by 576 Units/mg protein and the yield was 3.5% of the total activity of crude extracts. ${\beta}$-glucosidase specific activity was shown by 480 Units/mg protein and the yield was 2.9% of the total activity of crude extracts. The optimum temperature for ${\alpha}$-galactosidase was $60^{\circ}C$ and 43% of its original activity remained when it was treated at $80^{\circ}C$ for 30 min. For ${\alpha}$-galactosidase shows the optimum pH of 8.0 and is fairly stable between pH5.0 and pH9.0. The enzyme activity was increased in the presence of $Fe^{2+}$ and $Cu^{2+}$. The value of Km and Vmax for the enzyme were 0.98 mM and $1.81{\mu}$mole/min, respectively. The ${\beta}$-glucosidase has the optimum temperature of $50^{\circ}C$ and 46% of its original activity remained when it was treated at $80^{\circ}C$ for 30min. Its optimum pH of 7.0 and is fairly stable between pH5.0 and pH9.0. The enzyme activity was increased in the presence of $Fe^{2+},\;Co^{2+}$ and $Cu^{2+}$. The value of Km and Vmax for the enzyme were 1.24 mM and $6.81{\mu}$mole/min, respectively.

• Journal of Microbiology and Biotechnology
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• v.18 no.4
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• pp.695-703
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• 2008

A locally isolated thermophile, Geobacillus sp. SAB-40, producing thermostable extracellular amylase constitutively and an induced intracellular ${\beta}$-galactosidase was characterized and identified based on 16S rRNA sequencing. A phylogenetic analysis then revealed its closeness to Geobacillus stearothermophilus. To evaluate the effect of the culture conditions on the coproduction of both enzymes by G stearothermophilus SAB-40, a Plackett-Burman fractional factorial design was applied to determine the impact of twenty variables. Among the tested variables, $CaCI_2$, the incubation time, $MgSO_4{\cdot}7H_2O$, and tryptone were found to be the most significant for encouraging amylase production. Lactose was found to promote ${\beta}$-galactosidase production, whereas starch had a significantly negative effect on lactase production. Based on a statistical analysis, a preoptimized medium attained the maximum production of amylase and ${\beta}$-galactosidase at 23.29 U/ml/ min and 12,958 U/mg biomass, respectively, which was 3-and 2-fold higher than the yield of amylase and lactase obtained with the basal medium, respectively.

• BMB Reports
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• v.34 no.5
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• pp.395-401
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• 2001

Schizosaccharomyces pombe gene encoding redox enzymes, such as thioltransferase (TTase) and thioredoxin (TRX), were previously cloned and induced by oxidative stress. In this investigation, their expressions were examined using $\beta$-galactosidase fusion plasmids. The expression of the two cloned genes appeared to be growth-dependent. The synthesis of $\beta$-galactosidase from the TTase-lacZ fusion was increased in the medium with the low glucose level, whereas it was significantly decreased in the medium without glucose or with galactose. It was also decreased in the nitrogen-limited medium. The synthesis of galactosidase from the TRX-lacZ fusion was unaffected by galactose or low glucose. However, it was lowered the absence of glucose. The synthesis of $\beta$-galactosidase from the TTase-lacZ fusion was shown to be enhanced in a higher medium pH. Our findings indicate that S. pombe TTase and TRX genes may be regulated by carbon and nitrogen sources, as well as medium pH.

• Applied Biological Chemistry
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• v.39 no.1
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• pp.60-66
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• 1996

To investigate a possible application of three strains of lactic acid bacteria(strain No. 49. No. 61. No. 75) from kimchi in milk fermentation industry, the optimal condition for production of intracellular ${\beta}-galactosidase$ from Lactobacillus(L.) plantarum and its enzymatic properties were examined. The preferable carbon source of the medium for strain No. 49 in production of ${\beta}-galactosidase$ was MRS broth with 1.0% lactose instead of dextrose of pH 65. for strain No. 75 with 1.0% galactose and for strain No. 61 with 3.0% lactose at pH 7.5, respectively. The maximum enzyme production from strain No. 49, No. 75 was observed after 48 hours culture at $30^{\circ}C$ in a medium containing the appropriate carbon source, from strain No. 61 after 48 hours culture at room temperature. The optimum temperature for ${\beta}-galactosidase$ activity from L. plantarum was $60^{\circ}C$ for strain No. 49, $37^{\circ}C$ for strain No. 61 and $50^{\circ}C$ for strain No. 75, respectively. The heat stability of enzyme activities for all three strains remained 90% at $45^{\circ}C$. The optimal pH was pH 6.5 and enzyme activities were most stable at pH for all three bacteria.

• Microbiology and Biotechnology Letters
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• v.11 no.1
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• pp.59-66
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• 1983

Cultural conditions for the production of extracellar $\beta$-galactosidase by Loctobacillus sporogenes, a spore forming lactic acid bacterium, were investigated with shaken flask and jar fermenter cultures. The fermentation medium giving maximum $\beta$-galactosidase yield was found to consist of 1 % lactose as a carbon source, 1.5% peptone as an organic nitrogen source. 0.2% ammonium sulfate as an inorganic nitrogen source, 0.8% ammonium phosphate dibasic as a phosphorus source, and 0.05% potassium chloride and 0.001% ferric chloride as mineral source. Optimal initial pH of the medium was 7.0 and the highest enzyme excretion was observed after 40 hours of cultivation at 37$^{\circ}C$. In this experiment, the 500$m\ell$ conical flask containing 50-200$m\ell$ of medium was shaken at 140 strokes per minute with 7cm amplitude in a reciprocating shaker. The maximum enzyme value attained was 38 U/$m\ell$ of the culture broth which was found to be slightly higher than the highest intermolecular enzyme activity (30 U/$m\ell$) observed after 24 hours of incubation. In the fermentor culture, the fermentation profile was shown to be similar to that observed in the shaken flask experiment. But the maximum extracellular enzyme activity was 45 U/$m\ell$ to be even higher than the value obtained with the shaken flask culture.