• 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.

• Journal of Microbiology and Biotechnology
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• 제25권8호
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• pp.1291-1298
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• 2015

Five different polymer nanofibers, namely, polyaniline nanofiber (PANI), magnetically separable polyaniline nanofiber (PAMP), magnetically separable DEAE cellulose fiber (DEAE), magnetically separable CM cellulose fiber (CM), and polystyrene nanofiber (PSNF), have been used for the immobilization of lactase (E.C. 3.2.1.23). Except for CM and PSNF, three polymers showed great properties. The catalytic activities (k_cat) of the free, PANI, PAMP, and magnetic DEAE-cellulose were determined to be 4.0, 2.05, 0.59, and 0.042 mM/min·mg protein, respectively. The lactase immobilized on DEAE, PANI, and PAMP showed improved stability and recyclability. PANI- and PAMP-lactase showed only a 0-3% decrease in activity after 3 months of vigorous shaking conditions (200 rpm) and at room temperature (25℃). PANI-, PAMP-, and DEAE-lactase showed a high percentage of conversion (100%, 47%, and 12%) after a 1 h lactose hydrolysis reaction. The residual activities of PANI-, PAMP-, and DEAE-lactase after 10 times of recycling were 98%, 96%, and 97%, respectively.

• Asian-Australasian Journal of Animal Sciences
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• 제18권11호
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• pp.1617-1622
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• 2005

The activity of brush border enzymes (sucrase, lactase and maltase) in the piglet small intestine was evaluated as well as piglet performance during the weaning period in the present study. There were two treatment groups: Piglets of six litters were fed dry feed plus milk replacer (Group M) and of six litters fed dry pelleted feed (Group C). One piglet from each litter was sacrificed on day 3 before weaning, and day 3, 10 and 17 postweaning, respectively. Providing milk replacer caused an increased piglet live weight at weaning (p<0.001) and until termination of the experiment (p<0.001). A slightly higher (p<0.16) level of protein was measured in the jejunum of group M piglets as compared with group C piglets. Before weaning the activity of lactase was high in the jejunum of group C piglets. The activity of lactase in the jejunum was lowered in the jejunum of group C piglets and in distal jejunum of group M piglets during the postweaning period as compared with pre-weaning period (p<0.05). Lowered activity of lactase in the distal jejunum of piglets was found at day 10 and 17 postweaning, respectively. No treatment differences were found in the activity of lactase in the piglet jejunum. No treatment differences were seen in the activity of maltase and sucrase in the piglet jejunum also. However, weaning caused a higher activity of sucrase in the distal jejunum of group M piglets as compared with pre-weaning period. In conclusion, providing milk replacer to piglets caused an improved growth performance. Feeding milk replacer did not influence the activity of lactase, maltase and sucrase in the jejunum of piglets. Weaning resulted in a markedly lowered activity of lactase, while no dramatic changes in the activity of maltase took place during the period around weaning.

• 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.

• 한국축산식품학회지
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• 제27권4호
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• pp.496-500
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• 2007

The lactase activities of nine species of lactic acid bacteria were compared using the chromogenic substrate, $o-nitrophenyl-{\beta}-D-galactopyranoside$. Streptococcus thermophilus KACC 91147 had the highest lactase activity among a total of thirty strains of Lactobacillus and S. thermophilus tested, including commercial strains. S. thermophilus KACC 91147 released $0.30{\pm}0.12\;mg/mL$ of galactose in treated milk A ($10^7\;CFU/mL$) and $6.49{\pm}0.38\;mg/mL$ in treated milk B ($10^9\;CFU/mL$ milk) over 2 hours. In milk tolerance tests, the blood glucose level (BGL) of 6 volunteers (2 males and 4 females) clinically diagnosed as lactose intolerant increased 3.0 mg/dl after drinking milk A, but a significant (p<0.05) additional increase of $11.2{\pm}4.18\;mg/dl$ was found after drinking milk B. This result suggests that the addition of S. thermophilus KACC 91147 cells into milk aids the digestion of lactose in milk and ameliorates the symptoms of lactose-intolerant individuals due to the activity of lactase from the lactic streptococci.

• Journal of Dairy Science and Biotechnology
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• 제27권2호
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• pp.55-62
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• 2009

우유는 110여 종에 달하는 다양한 영양소를 골고루 함유하고 있어 성장기 어린이들에게 영양학적으로 완전에 가까운 식품으로 알려져 있다. 전세계 인구의 75% 정도가 유전적으로 유당을 소화시키는 능력이 부족한 이른바 유당불내증 때문에 우유 소비를 가로막는 최대 장애요인이 되고 있다. 우유의 섭취 부족으로 인하여 영양소의 결핍이 초래되어 골다공증, 고혈압, 결장암의 발병율이 증가하고 있는 추세이다. 식품 중에 함유된 유당이 체내에 흡수되려면 소장내에서 가수분해가 일어나야 한다. 유당을 분해하는 효소를 락타아제 또는 $\beta$-갈락토시다아제라 한다. 유당의 가수분해로 생성되는 포도당은 혈당이나 에너지원으로 이용되지만, 갈락토오스는 유아의 뇌조직 성분인 당지질의 합성에 이용된다. 임상적으로 유당불내증은 breath hydrogen production test로 진단할 수 있다. 유당불내증은 유럽인 25%, 히스패닉계, 인도인, 흑인 및 유대인 50~80%, 아시아인과 아메리카 인디언의 경우 거의 100% 생기는 것으로 보고되었다. 성인의 경우 유당을 가수분해할 수 있는 능력 즉, 락타아제 지속성(lactase persistence)이 북유럽 인들에게는 흔한 표현형 인 반면 그 외 지역에서는 락타아제 비지속성(lactase non-persistence) 또는 성인형 저유당분해 효소증(adult-type hypolactasia)이 가장 흔하게 나타난다. 유전자 분석 결과 인간의 락타아제 유전자의 5' 말단으로부터 13910째 염기의 다혈성(C/T-13910 single nucleotide polymorphism)이 락타아제 지속성과 관련이 있다는 사실이 밝혀졌다. 유당불내증을 완화하기 위해서는 1회분으로 한 컵 이하, 타 식품과 함께 마실 것, 탈지유보다는 전지유, 요구르트 유산균과 함께 마실 것 등이 권장되며 위배출지연도 유당불내증 개선에 효과적이다. 기존 유당불내증 진단법은 지나치게 많은 유당을 투여함으로써 유당불내증 비율이 과장되어 있으므로 정확한 진단법의 개발이 필요하다. 발효유 제품이나 저유당 제품을 개발하면 유당불내증으로 인한 유제품의 소비 부진을 크게 해소할 수 있을 것으로 판단된다.

• Asian-Australasian Journal of Animal Sciences
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• 제15권12호
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• pp.1808-1812
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• 2002

The present study was carried out to examine the efficiency of microcapsules and a stability of lactase in vitro in the simulated gastric and intestinal conditions. As a coating materials, medium-chain triacylglycerol (MCT) and polyglycerol monostearate (PGMS) were used. The highest efficiency of microencapsulation was found in the ratio of 15:1 as coating to core material with both MCT (91.5%) and PGMS (75.4%). In a subsequent experiment, lactose content was measured to study a microcapsule stability. Lysis of microcapsules made by MCT in simulated gastric fluid was proportionally increased such as 3% in pH 5 and 11% in pH 2 for 20 min incubation. In the case of PGMS microcapsulation, 11-13% of lactose was hydrolyzed at 20 min in all pHs and also very little amount (less than 3%) of lactose was hydrolyzed after 20 min in all pHs. The highest percentages of lactose hydrolysis in MCT and PGMS microcapsules were 68.8 and 60.8% in pHs 7 and 8 during 60 min, respectively. Based on our data, the lactase microcapsules seemed to be stable when they stay in the stomach, and hydrolyzed rapidly in small intestine where the bile acid was excreted.

• Journal of Dairy Science and Biotechnology
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• 제39권1호
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• pp.20-26
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• 2021

Lactase (β-galactosidase) is abundant in the small intestine during early childhood and gradually decreases with age. Lactic acid bacteria (LAB) present in yogurt could survive in the stomach, and lactase produced by these LAB can aid in lactose breakdown in the small intestine, thereby reducing lactose intolerance. This study aims to provide preliminary data for development of lactose-free yogurts for the elderly, and investigate the effect of lactose-hydrolyzed milk on the growth of starter cultures. The pH during yogurt fermentation using lactose-free milk was slightly higher at 2 and 4 h of incubation, but reached 4.5 at the end of incubation, similar to that of the yogurt prepared from regular milk. The number of viable cells of Streptococcus thermophilus reached 108 CFU/mL after 2 h of incubation and increased to 109 CFU/mL after 4 h of incubation. During yogurt fermentation, the viable cells of Lactobacillus species and Bifidobacterium longum did not affect lactose hydrolysis. Although lactose-hydrolyzed milk did not promote the growth of starter cultures, manufacturing yogurt with lactose-free milk could be beneficial for the intestinal health of lactose-sensitive elderly.

• Journal of Dairy Science and Biotechnology
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• 제34권4호
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• pp.231-237
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• 2016

Kefir is a probiotic food. Probiotics have shown to be beneficial to health, and are currently of great interest to the food industry. Hence, this study was carried out to evaluate the lactase activity of kefir-isolated lactic acid bacteria. Three strains, Lactobacillu kefiri DH5 isolated from the kefir grains and Bifidobacterium animalis subsp. Lactis and Bifidobacteria longum 720, commercial probiotic LAB, were fermented in 10% reconstituted nonfat dry milk suspensions and incubated at $37^{\circ}C$ for 48 h, and then analyzed for various saccarides by HPLC. The results showed that changes in the concentrations of lactose and galactose were significantly decreased and increased, respectively (p<0.05), but all 3 probiotic strains tested in this study showed no increase in glucose concentration during 48 h of incubation. Both DH5 and BL720 showed high lactase activities (p<0.05), whereas BLC exhibited the lowest activity. Additionally, all three lactic acid bacteria showed high tagatose, but did not show high xylose and sedoheptulose. Finally, DH5, a kefir-isolated LAB, may have similar characteristics and properties to typical Bifidobacterium spp. and showed higher lactase activity than commercial Bifidobacterium spp.

• Journal of Dairy Science and Biotechnology
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• 제33권4호
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• pp.263-269
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• 2015

본 연구에서는 다양한 낙농 발효유제품에 있어서 다양한 유기산과 탄수화물의 분석이 HPLC 방법으로 동시에 잘 진행되었다. 이 방법의 주요 장점으로는 (1) 간단한 시료 준비와 단일 추출이 가능, (2) 유동상(mobile phase)과 추출 용매에 있어서 편리성과 경제성, (3) 추출제와 유동상에 사용되는 황산의 농도가 매우 낮기에 친환경적인 실험, (4) 하나의 샘플당 30분 안에 HPLC의 분석이 완료 가능, (5) 다양한 농도범위에서도 분석 및 정량 가능, (6) 몇 가지 물질을 제외하고 모든 화합물의 변동계수가 낮음, 그리고 (7) 회수율이 대체적으로 높다는 것이다. 또한, $60{\sim}65^{\circ}C$에서 분석 column을 사용하였기에 column 효율면에서도 손실이 극히 적은 것으로 관찰되었다. 따라서 본 실험에서 분석된 다양한 낙농 발효유제품에서의 acid, 탄수화물, diacetyl, acetoin 등이 HPLC 방법은 매우 신속하고 정확하게 검출되었다. 하지만 다양한 향미의 변화를 최소화하면서 다양한 향미를 효과적으로 추출할 수 있는 HPLC 방법들이 지속적으로 개발되어야 다양한 낙농 발효유제품의 품질개선에 기여할 것으로 사료된다. 왜냐하면 향미 성분의 분석은 성분을 정성하는 것뿐만 아니라, 정성된 화합물이 그 식품의 향에 어느 정도 기여하는지를 중요하게 결정하기 때문이다. 또한 본 실험에 사용된 HPLC 방법으로 다양한 유산균주의 일정한 배양시간동안 유당분해효소(lactase) 능력이 조사 되었는데, 각 Probiotic lactic acid bacteria마다 다양한 유당분해효소(lactase)의 능력을 보였으며, 같은 균종에서도 차이가 있는 것으로 나타났다. 본 실험에서는 Bifidobacterium spp.가 Lactobacillus spp.와 Streptococcus spp.보다 높은 유당분해효소(lactase)의 능력을 보였다. 하지만 유산균주의 유당분해효소 능력을 정확하게 파악하기 위해서는 각 Probiotic lactic acid bacteria의 exo- 및 endo-lactase에 대한 연구가 추가적으로 필요하다. 결론적으로 다양한 낙농발효식품 및 기타 발효식품 등에서 HPLC 방법으로 동시에 유기산과 탄수화물 분석이 공인방법으로써 인정되기 위해서는 더 많은 기초자료 확립과 재현성에 관한 지속적인 연구가 진행되어야 할 것으로 사료된다.