• Journal of Microbiology and Biotechnology
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• 제5권1호
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• pp.6-13
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• 1995

A gene coding for the $\beta$-galactosidase (lactase) of Kluyveromyces tragilis UCD 55-55 was isolated by complementation in Escherichia coli YMC9. From the plasmid library made from Sau3A-digested chromosomal DNA, one positive clone was selected. The cloned gene for $\beta$-galactosidase was on 7.3 kilobase pair DNA fragment, and a slightly low level of $\beta$-galactosidase enzyme activity was detecied in E. coli. It was also confirmed that the cloned gene comes from K. tragilis by DNA-DNA hybridization and immunochemical blotting experiments. In order to construct a new yeast strain having the metabolic ability for lactose, the cloned gene for K. tragilis $\beta$-galactosidase was inserted in yeast vector YEp24 and YRp17, and transformed into Saccharomyces cerevisiae YNN27 and Ml-2B. The yeast transformants showed the nearly the same $\beta$-galactosidase productivity as level of K. tragilis when uninduced, but these could not utilize lactose as a sole carbon source, presumably due to the lack of lactose transport system. Nevertheless, a slightly higher ethanol productivity was achieved by these transformants than S. cerevisiae or K. tragilis, in the medium containing glucose and lactose.

• 한국식품영양과학회지
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• 제21권1호
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• pp.60-63
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• 1992

본 실험은 yoghurt와 yoghurt culture에서의 ${\beta}-galactosidase$의 활성도를 측정하였다. Liquid medium에서 자란 yoghurt culture의 ${\beta}-galactosidase$의 활성도는 S. thermophilus가 Lactobacillus에 비해 $5{\sim}10$배 높은 것으로 나타났으며 제조 yoghurt를 저장온도에 따른 차이를 실험한 결과 냉장보관$(4^{\circ}C)$시 한달 정도까지는 효소활성도와 생균수를 보전함을 볼 수 있었으나 실온저장시는 5일 정도 밖에 효소활성도와 생균수를 유지하지 못했다. 실험을 통해 ${\beta}-galactosidase$ activity는 생균수와 비례함을, 특히 S. thermophilus의 생균수와 비례함을 알수 있었다. 시판 yoghurt와의 비교실험에서는 활성도와 생균수에 있어서 반정도의 수준도 못 미침을 알수 있었는데 이는 다음호에 계속될 여러 요인들에 기인함을 짐작케 한다.

• Journal of Microbiology and Biotechnology
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• 제18권4호
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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.

• 한국축산식품학회지
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• 제27권3호
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• pp.329-336
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• 2007

This study was designed to find the optimum conditions for converting isoflavone glycoside to aglycone by ${\beta}-galactosidase$. Three different forms of the enzyme were tested and the optimum enzyme concentration, incubation temperature, pH, and incubation time were determined. Before treatment with enzyme, isoflavone contained 89.4% glycoside including daidzin, glycitin and genistin, and only 10.6% aglycone including daidzein, glycitein and genistein. Among the enzymes tested, the highest rate of isoflavone hydrolysis to aglycone, 35%, was observed when 3 unit/g Fungal Lactase (Amano Enzyme) was used. Higher incubation temperatures resulted in a higher rate of hydrolysis along with a greater loss of isoflavone mass. Therefore, body temperature $(37^{\circ}C)$ may be adequate for isoflavone conversion, with 44.9% hydrolysis and less than 10% loss of mass. As expected, a higher amount of aglycone was produced at pH 7 compared with other pH values. During 5hr of incubation, the conversion of glycoside to aglycone increased dramatically from 0 to 1hr, and plateaued thereafter. In addition, commercial soy-based milk was hydrolyzed more effectively with ${\beta}-galactosidase$ when incubated for 5hr. Based on the above results, the optimum conditions for isoflavone hydrolysis by ${\beta}-galactosidase$ were for 3 hr at $37^{\circ}C$, pH 7 with 3 unit/g Fungal Lactase (Amano Lactase), yielding an average total amount of aglycone ranging from 40 to 47%.

• Asian-Australasian Journal of Animal Sciences
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• 제15권4호
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• pp.596-601
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• 2002

The present study was designed to examine the effect of ozone treatment in microencapsulated ${\beta}$-galactosidase on inactivation of the enzyme and sterilization of microorganism. The efficiency was the highest as 78.4% when the ratio of polyglycerol monostearate (PGMS) was 15:1. Activities of lactase remaining outside the capsule were affected by ozone treatment. With the increase of ozone concentration and duration of ozone treatment, the activity reduced significantly. In sensory aspect, with 2% microcapsule addition, no significant difference in sweetness was found compared with a market milk during 12 d storage. Above result indicated that the additional washing process of lactase was not necessary to inactivate the residual enzyme. In a subsequent study, the vegetative cells of microorganisms were completely killed with 10 ppm for 10 min treatment by ozone. The present study provides evidence that ozone treatment can be used as an inactivation and a sterilization process. In addition, these results suggest that acceptable milk products containing lactase microcapsules made by PGMS can be prepared with ozone treatment.

• 미생물학회지
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• 제42권3호
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• pp.216-222
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• 2006

[ ${\beta}-Galactosidase$ ] (lactase)를 생산하는 유산균을 선별하고 그들이 생산하는 ${\beta}-galactosidase$의 특성을 조사하였다. X-gal이 포함된 MRS배지에서 푸른색을 나타내는 약 100여개의 콜로니를 김치로부터 분리하였다. 그들 중 ET-1과 LA-12 두 균주를 최종 선별하였으며, 16S rDNa 염기서열 분석을 통해 각 각 Lactobacillus fermentum과 L. acidophilus와 상동성이 높은 Lactobacillus 속으로 동정되었다. 선별된 두 균주는 높은 ${\beta}-galactosidase$ activeity와 우수한 생존력을 나타내었으며, 이들이 생산하는 ${\beta}-galactosidase$의 최적 활성조건을 조사한 결과, 반응온도 $55^{\circ}C$에서 가장 높은 활성을 나타내었으며, 반응 pH의 경우 ET-1은 pH 5.5에서 최고 활성을 나타내었고, LA-12는 pH7.0에서 가장 높은 활성을 보였다. 선별 유산균 ET-1과 LA-12는 위액과 담즙산액에 대해 우수한 내성을 나타내었다. 두 개의 선별 균주는 인공위액에 3시간 배양 후 초기 생균수와 비교하여 생균수 변화가 거의 없었고, 담즙산액으로 사용한 0.3% oxgall에서 24시간 반응 후에도 1 log cycle 정도 감소된 $10^8CFU/ml$의 생균수를 유지하였다. 이러한 결과를 바탕으로 유산균 ET-1과 LA-12는 유제품 산업에 이용 가능성이 크다고 판단된다.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1996년도 춘계학술대회
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• pp.283-283
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• 1996

The hydrolyzed-lactose milk for the lactase-deficient subject is sweeter than whole milk, and some subjects dislike its taste. To overcome this shortcoming the dried liposomes containing ${\beta}$-galactosidase to digest lactose in milk after drinking were prepared and examined the possible application of this dried liposomes to the lactase-deficient subjects. To improve the stability of conventional liposome suspension, the dried liposomes in the presence of trehalose were prepared by the dehydration-rehydration vesicles method. Small unilamellar vesicles, prepared with egg phosphatidyl cholesterol, and cholesterol, were mixed with ${\beta}$-galactosidase solution and then ;up[jo;ozed. The freeze-dried liposome was rehydrated and centrifuged. The resultant multilamellar vesicles were mixed with trehalose(4g/g lipid) and then lyophilized to produce final dried liposome. Trehalose increased the entrapping efficiency of liposomes by 3 fo1d compared to the liposomes without trehalose (13% vs. 46%).

• 한국축산식품학회지
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• 제33권5호
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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.

• 미생물학회지
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• 제42권1호
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• pp.40-46
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• 2006

우유에 풍부하게 존재하는 유당은 galactose와 포도당의 $\beta(1\rightarrow4)$ glycosidic 결합으로 구성되어 있고, 인간에서 이를 가수분해하는 효소는 lactase, 세균에서는 $\beta-galactosidase$로 알려져 있다. Lactase의 활성이 낮은 사람이 우유를 섭취했을 경우 일시적인 설사를 일으키고 때로는 만성적인 대감의 염증으로 인한 만성설사의 원인이 되기도 한다. 겨울철에 젖소를 사육하는 축사 주변에서 저온에서 생육하는 세균 AS-20을 분리하여 $\beta-galactosidase$ 활성을 갖는 균주를 선별하고 온도별로 분리균의 성장을 조사하였다. 그 결과 대장균이 자라지 못하는 $10^{\circ}C$에서도 분리된 AS-20은 생육이 가능하였고 생육 최적온도는 $30^{\circ}C$이였으며 이 온도에서 세대시간은 60여분 이었다. AS-20의 생화학적 특성을 bioMerieux Vitek Gram negative identification card (GNI+)로 조사한 결과 포도당을 발효, 산화시켰으며 유당, maltose, mannitol, xylose, L-arabinose 등을 이용하여 97% Hafnia alvei, 2% Escherichia coli로 동정되었다. Polymerase chain reaction으로 16S rRNA유전자를 증폭하여 1,426 bp의 염기서열을 결정하여 기존에 보고된 유전자들과의 유사도를 조사한 결과 분리된 균주 AS-20은 Hafnia alvei와 99%의 염기서열상 동성을 보였다. 이러한 결과는 BioMerieux Vitek Gram negative identification card 키트로 동정한 결과와 일치하였다. AS-20을 $10^{\circ}C,\;20^{\circ}C,\;30^{\circ}C$에서 배양하면서 $\beta-galactosidase$ 활성을 조사한 결과 저온인 $10^{\circ}C$와 $20^{\circ}C$에서 배양하였을 때에 배양최적 온도인 $30^{\circ}C$에서 배양했을 때 보다 1.5 배 정도 높은 효소활성을 보여주었으며, $30^{\circ}C$에서 배양된 대장균 보다 6배 이상의 효소활성을 보여 주어 저온조건에서 분리균의 효소생산이 비교적 높은 것으로 판단되었다.

• 한국식품과학회지
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• 제21권6호
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• pp.795-799
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• 1989

효모 Kluyveromyces fragilis SKD-7001로부터 정제한 ${\beta}-galactosidase$를 이용하여 시유의 유당을 가수분해 시키고, 이 효소의 처리유로 요구르트를 제조하여 이들의 이화학적 성분의 변화를 검토하였다. 효소의 농도를 3.0units/ml 처리시 120분간 $40^{\circ}C$에서 반응시켰을 때 유당의 가수분해율이 75.0%를 나타내었고, 6.0units/ml 처리시 120분간 반응시켰을 때에는 94.6%이었다. ${\beta}-galactosidase$의 처리유로 요구르트를 제조하여 pH, 산도, 유산균의 생균수를 정토해 본 결과, $40^{\circ}C$에서 8시간 매양하였을 때 pH4.1, 산도 1.04% 그리고 유산균의 생균수는 각각 Str. thermophilus가 $6.5{\times}10^8/ml$, L. bulgaricus가 $8.9{\times}10^8/ml$를 나타내었다. 총 아미노산을 분석해 본 결과 총 함량은 대조 요구르트가 2.63%, LH 요구르트가 2.19%로써, LH 요구르트보다 대조 요구르트 함량이 더 높은 것으로 나타났다. 유리 아미노산을 분석해 본 결과 총 함량은 시유의 경우 대조 요구르트가 26.95mg%, LH 요구르트가 17.55mg%로써, LH 요구르트보다 대조 요구르트의 함량이 더 높은 것으로 나타났다. 지방산에 있어서는 LH 요구르트가 대조 요구르트보다 caprylic, capric, lauric acid 등의 저급 지방산 함량이 약간 높은 경향을 보였고, 고급 지방산 중에서는 palmitic acid의 함량이 가장 많은 것으로 나타났다.