• Microbiology and Biotechnology Letters
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• v.20 no.6
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• pp.647-654
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• 1992

In order to study the physiological properties of the intestinal bacteria, we isolated the intestinal bacteria of Koreans and tested the enzymatic patterns. Isolated Bifidobacterium sp. Int-57 had the higher activity of $\alpha$-glucosidase, $\beta$-glucosidase, $\alpha$-galactosidase, $\beta$-galactosidase. $\beta$-xylosidase and $\alpha$-arabinofuranosidase than other intestinal microorganisms. The effect of the carbon sources on the production of each enzymes of Bijidobacterium sp. Int-57 was investigated. The most suitable carbon source for the production of $\beta$-glucosidase was maltose, for a-glucosidase cellobiose, for $\alpha$-galactosidase raffinose, for $\beta$-galactosidase lactose, and for $\beta$-xylosidase and $\alpha$-arabinofuranosidase xylose, respectively. In addition, we investigated the optimal conditions and pH stability of each crude enzymes. The optimal condition of a-glucosidase was pH 6.0 and $40^{\circ}C$. that of Jj-glucosidase pH 7.0 and 50oe, that of $\beta$-galactosidase pH 7.0 and $50^{\circ}C$, that of $\beta$-xylosidase pH 6.0 and $40^{\circ}C$ , and that of $\alpha$-arabinofuranosidase pH 5.0 and $50^{\circ}C$. respectively. a-Glucosidase was stable at pH 4.0-9.0. Jj-glucosidase at pH 4.0-7.0. $\beta$-galactosidase at pH 4.0-9.0, $\beta$-xylosidase at pH 4.0-6.0, and /3-arabinofuranosidase at pH 7.0-9.0, respectively.

• Food Science of Animal Resources
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• v.33 no.5
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• pp.565-571
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• 2013

An enzyme ${\beta}$-galactosidase or ${\beta}$-galactohydrolase [EC3.2.1.23], commonly called lactase, mediates galacto-oligosaccharide (GOS) synthesis under conditions of high substrate concentrations. Also, lactase hydrolyzes ${\beta}$($1{\rightarrow}4$) lactose into glucose and galactose, the latter is successively transferred to free lactose to make various oligosaccharides via transgalactosylation. GOS is non-digestible to human digestive enzymes and has been used as a functional prebiotics. Among the 24 lactic acid bacteria (LAB) strains used, Lactobacillus paracasei YSM0308 was selected based on its exhibition of the highest ${\beta}$-galactoside hydrolysis activity, and the crude lactase was prepared for examination of reaction conditions to affect the GOS synthesis. Lactase activity was measured with a spectrophotometer using ONPG (o-nitropheyl ${\beta}$-D-galactopyranoside) method. Lactase activity was not detected in the culture supernatant and was mostly present in the cell pellet after centrifugation. Activity of the crude lactase preparation ranges from102 to 1,053 units/mL, with the highest activity determined for L. paracasei YSM0308. Optimal conditions for GOS synthesis are as follows: concentration of whey powder, pH, temperature, and time were 30%, pH 6.5-7.0, $30^{\circ}C$, and 4 h, respectively. The final GOS concentration was 19.41% (w/v) by the crude YSM0308 lactase, which was obtained from strain YSM0308 grown in the 10% (w/v) reconstituted whey-based medium.

• Korean Journal of Microbiology
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• v.27 no.3
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• pp.188-193
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• 1989

Gene for lactose catabolism in Lactobacillus casei SW-M1 was encoded by a 60Kb metabolic plasmid. A derivative of only 10kb, pPlac 15 of recombinant plasmid, was constructed by introducing into pBR322 and was cloned into E. coli using restriction endonuclease Pst I. A 10kb insery DNA in plasmid pBR322 was identified as a gene encoded phospho-$\beta$-galactosidase by the determination of enzyme activity. Phospho-$\beta$-galactosidase was apparently expressed in E. coli. The enzyme activities of cell-free extract from transformant E. coli HB101 carrying pPLac 15 DNA were not different from that of L. casei as a donor strain on the basis of enzyme properites. However, specific activity of phospho-$\beta$-galactosidase in the cloned strain with Lac $Y^{-}$ phenotype of E. coli HB101 was lower than that in L. casei strain.

• Reproductive and Developmental Biology
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• v.39 no.4
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• pp.97-104
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• 2015

Glucose is universal and essential fuel of energy metabolism and in the synthesis pathways of all mammalian cells. Glucose is the one of the major precursors of lactose synthesis using glycolysis result in producing milk fat and protein. During the milk fat synthesis, lipoprotein lipase (LPL) and CD36 are required for glucose uptake. Various morecules such as acyl-CoA synthetase 1 (ACSL1) activity of acetyl-CoA synthetase 2 (ACSS2), ACACA, FASN AGPAT6, GPAM, LPIN1 are closely related with milk fat synthesis. Additionally, glucose plays a major role for synthesizing lactose. Activations of lactose synthesize enzymes such as membranebound enzyme, beta-1,4-galactosyl transferase (B4GALT), glucose-6-phosphate dehydrogenase (G6PD) are changed by concentration of glucose in blood resulting change of amount of lactose production. Glucose transporters are a wide group of membrane proteins that facilitate the transport of glucose over a plasma membrane. There are 2 types of glucose transporters which consisted facilitative glucose transporters (GLUT); and sodium-dependent transport, mediated by the Na+/glucose cotransporters (SGLT). Among them, GLUT1, GLUT8, GLUT12, SGLT1, SGLT2 are main glucose transporters which involved in mammary gland development and milk synthesis. However, more studies are required for revealing clear mechanism and function of other unknown genes and transporters. Therefore, understanding of the mechanisms of glucose usage and its regulation in mammary gland is very essential for enhancing the glucose utilization in the mammary gland and improving dairy productivity and efficiency.

• Journal of Life Science
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• v.23 no.12
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• pp.1471-1476
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• 2013

The present study deals with the immobilization of Kluyveromyces lactis ${\beta}$-galactosidase on a weak ionic exchange resin (Duolite A568) as polymer support. ${\beta}$-Galactosidase was immobilized using the adsorption method. A kinetic study of the immobilized enzyme was performed in a packed-bed reactor. The adsorption of the enzyme followed a typical Freundlich adsorption isotherm. The adsorption parameters of k and n were 14.6 and 1.74, respectively. The initial rates method was used to characterize the kinetic parameters of the free and immobilized enzymes. The Michaelis-Menten constant ($K_m$) for the immobilized enzyme (120 mM) was higher than it was for the free enzyme (79 mM). The effect of competitive inhibition kinetics was studied by changing the concentration of galactose in a recycling packed-bed reactor. The kinetic model with competitive inhibition by galactose was best fitted to the experimental results with $V_m$, $K_m$, and $K_I$ values of 46.3 $mmolmin^{-1}mg^{-1}$, 120 mM, and 24.4 mM, respectively. In a continuous packed-bed reactor, increasing the flow rate of the lactose solution decreased the conversion efficiency of lactose at different input lactose concentrations. Continuous operation of 11 days was conducted to investigate the stability of a long-term operation. The retained activity of the immobilized enzymes was 63% and the half-life of the immobilized enzyme was found to be 15 days.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2004.06a
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• pp.328-333
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• 2004

• The Journal of Natural Sciences
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• v.8 no.2
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• pp.57-61
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• 1996

The $\beta$-glucosidase enzyme was purified from E. coli carrying Cellulomonas fimi $\beta$-glucosidase gene. SDS-PAGE and analytical gel filtration revealed that molecular weight of this enzyme was 56,000 dalton and consisted of a single polypeptide.Inhibition caused by heavy metals and activation by dithiothreitol suggest the existence of essential thiol group in the enzyme. The enzyme was not active on maltose (glucose $\alpha$-1,4-glucose) which has a $\alpha$-linkage, whereas it was active on lactose (glucose $\beta$-1,4-glucose), PNPG (p-nitrophenyl $\beta$-D-glucopyranoside) and PNPC (p-nitrophenyl $\beta$-D-cellobioside), although its reaction rates were different.

• Journal of the Korean Society of Food Science and Nutrition
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• v.25 no.1
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• pp.150-156
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• 1996

유산균의 균주개량방법의 일환으로 photoplast fu-sion 기법과 electrofusion법을 이용하여 protease 활성, lipase 활성, 내열성, 내산성 등이 우수한 L. bulgaricus와 bacteriocin을 생산하는 L. acidophilus 간의 융합시켜 얻은 융합주들의 생리학적 성질을 검토하였다. 산 생성능, 내열성, 내산성 protease, lipase 활성 등 L. bulgaricus가 L. acidophilus보다 우수하였다. L. bulgaricus는 lactose와 sorbos를 이용하였으나. mal-tose 와 dorbitol을 이용하지 못하는 반면 , L. acidophilus는 matose를 이용하고 lactose 와 sorbose를 이용하지 못하였다. 융합주 가운데서는 367810번이 모균주의 발효능 특성을 함께 지님으로서 재조합체임을 확인 할 수 있었으며, sorbitol의 경우 모균주에서는 발효능이 전혀 나타나지 않았음에도 불구하고 융합주 4, 7번이 발효능을 나타내어 융합과정에서 새로운 형질을 획득하기도 한다는 사실을 알 수 있었다. Lactase 활성은 모균주 모두 높은 $\beta-galactosidase$ 활성을 보였으나, $phospho-\beta-galactosidase$ 활성은 거의 없었으며 융합주들도 다소 차이는 있었지만 모균주와 유사한 효소활성을 나타내었다. 발효에 있어서 key enzyme으로 작용하는 protease. lipase 등의 효소활성도 모균주의 활성보다 우수한 융합주도 존재하였다.

• Journal of Microbiology and Biotechnology
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• v.16 no.7
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• pp.1026-1031
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• 2006

The optimization of the production of a fusion protein containing the antigenic domain 1 (AD-1) is of a great importance, considering its use in diagnostic tests. The fusion protein is produced by the fermentation of a recombinant strain of Escherichia coli containing the plasmid Mbg58, which expresses the AD-1 (aa 484-650) of human cytomegalovirus glycoprotein B as a fusion protein together with aa 1-375 of ${\beta}-galactosidase$. An important characteristic of promoters (lac and derivatives) used in recombinant protein production in E. coli is their inducibility. Induction by IPTG is widely used for basic research; however, its use in large-scale production is undesirable because of its high cost and toxicity. In this work, studies using different inducers and carbon sources for the production of a fusion protein containing the AD-l were performed. The results showed that lactose could be used as an inducer in the fermentation process for the production of this protein, and that expression levels could exceed those achieved with IPTG. The use of lactose for protein expression in E. coli should be extremely useful for the inexpensive, large-scale production of heterologous proteins in E. coli. Addition of sucrose to the fermentation medium improved the yield of recombinant protein, whereas addition of fructose or trehalose decreased the yield.

• 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.