• 한국미생물·생명공학회지
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• 제11권1호
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• pp.5-13
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• 1983

고온균의 생리적 특성자 내열기구에 관한 기초 자료를 얻고자 온천토양에서 $\beta$-galactosidase를 생산하는 고온균을 분리하여 동정하고 몇가지 주요한 생리적 특성을 검토하였으며 그의 균체 지방산 조성을 gas chromatography로 분석하여 다음과 같은 결과를 얻었다. 1. $\beta$-galactosidase를 생산하는 공시균주는 Thermus sp. 으로 동정되었다. 2. 분리선정한 균주의 최적 생육온도는 $65^{\circ}C$이었고, 37$^{\circ}C$에서 생육하지 않는 절대고온균 이었으며 최적pH는 6.5내외이었고 pH에 민감하였다. 3. NaCl에 대한 내성은 NaCl 1% 이상에서는 생육하지 못했다. 4. 항생물질에 대한 내성은 penicillin G는 10$\mu\textrm{g}$/$m\ell$, chloramphenicol은 0.5$\mu\textrm{g}$/$m\ell$ 이었다. 5. Vitamin 요구성은 Ca-pantothenate와 pyridoxine-HCI를 절대적 생육인자로 요구하였고niacin을 자극적 생육인자로 요구하였다. 6. 공시균주의 균체 지방산 조성은 palmitic acid 60.20%, lauric acid 11.80%, myristic acid 7.56%. behenic acid 4.25%, Capric acid 1.77%, stearic acid 2.13%, arachidic acid 1.53% 이었다.

• 한국식품과학회지
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• 제28권3호
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• pp.451-457
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• 1996

Bifidobacterium breve의 최적 배양 조건과 안정성을 증진시킬 수 있는 배양조건을 조사하기 위하여 pH와 L-cysteine HCI의 첨가효과를 조사하였다. 최대 균체수를 얻기위한 pH 조건은 pH $6.0{\sim}6.5$가 최적 배양 조검임을 알 수 있었다. 저장시의 최적 배양 조건은 $5.5{\sim}6.0$에서 가장 우수하였다. pH $5.5{\sim}6.0$에서 배양한 균주를 $4^{\circ}C$에서 저장시 25일 이후에 $2.4{\times}10^6/ml$와 $1.4{\times}10^6/ml$로 $10^6/ml$ 이상을 유지하여 저장시 안정성이 우수한 반면 pH 7.0에서 배양한 균주는 $2.4{\times}10^6/ml$로 저장시 생존력이 상당히 낮았으며 pH 5.5와 6.0에서 배양한 균주의 ${\beta}$-galactosidase 활력은 25일 저장 이후에도 $78{\sim}85%$를 유지한 반면 pH 7.0에서 배양한 균주의 ${\beta}$-galactosidase 활력은 급격히 감소하였다. L-cysteine HCI 첨가 효과는 0.05% 이상 첨가시 증식에 유리한 환경을 제공하였으며 과산화수소에 대한 내성은 $0.05{\sim}0.10%$ L-cysteine HCI을 첨가한 배이에서 증식한 B. breve가 우수하였다. Osmoprotectant를 첨가하여 배양시 증식과 동결건조시 균주의 안정성에 대한 효과를 조사해 본 결과 2mM betaine이나 2mM trehalose를 첨가시 증식에도 효과가 있었으며 osmoprotectant 역할을 함을 알 수 있었다. 또 한 동결건조에서도 betaine이나 trehalose를 첨가하여 배양된 B. breve는 거의 손상을 받지 않음을 알 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제29권11호
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• pp.1717-1728
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• 2019

The gene encoding β-galactosidase was cloned from Bifidobacterium longum RD47 with combinations of several bifidobacterial promoters, and expressed in B. bifidum BGN4. Among the recombinant bifidobacteria, BGN4+G1 showed the highest β-galactosidase level, for which the hydrolytic activity was continuously 2.5 to 4.2 times higher than that of BGN4 and 4.3 to 9.6 times higher than that of RD47. The β-galactosidase activity of BGN4+G1 was exceedingly superior to that of any of the other 35 lactic acid bacteria. When commercial whole milk and BGN4+G1 were reacted, BGN4+G1 removed nearly 50% of the lactose in the milk by the 63-h time point, and a final 61% at 93 h. These figures are about twice the lactose removal rate of conventional fermented milk. As for the reaction of commercial whole milk and crude enzyme extract from BGN4+G1, the β-galactosidase of BGN4+G1 eliminated 51% of the lactose in milk in 2 h. As shown below, we also compared the strengths and characteristics of the strong bifidobacterial promoters reported by previous studies.

• Journal of Microbiology and Biotechnology
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• 제27권3호
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• pp.598-609
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• 2017

Pediococcus pentosaceus ID-7 was isolated from kimchi, a Korean fermented food, and it showed high activity for lactose hydrolysis. The ${\beta}$-galactosidase of P. pentosaceus ID-7 belongs to the GH2 group, which is composed of two distinct proteins. The heterodimeric LacLM type of ${\beta}$-galactosidase found in P. pentosaceus ID-7 consists of two genes partially overlapped, lacL and lacM encoding LacL (72.2 kDa) and LacM (35.4 kDa). In this study, Escherichia coli MM294 was used for the production of LacL, LacM, and LacLM. These three types of recombinant proteins were expressed, purified, and characterized. The specific activities of LacLM and LacL were 339 and 31 U/mg, respectively. However, activity was not detected with LacM alone. The optimal pH of LacLM and LacL was pH 7.5 and pH 7.0, and the optimal temperature of LacLM and LacL was $40^{\circ}C$ and $50^{\circ}C$, respectively. The optimal temperature changes indicate that LacLM is able to achieve higher activity at a relatively lower temperature. LacLM was strongly activated by $Mg^{2+}$, $Mn^{2+}$, and $Zn^{2+}$, which was not true for LacL. Consistent with this, EDTA strongly inactivated LacLM and LacL, but the presence of reducing agents did not dramatically alter the activity. Taken together, multiple alignment of amino acid sequences and phylogenetic analysis results of LacL and LacM of P. pentosaceus ID-7 suggest the evolution of LacL into LacLM and that the use of divalent metal ions results in higher activity.

• 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%).

• Asian-Australasian Journal of Animal Sciences
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• 제20권7호
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• pp.1015-1022
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• 2007

The spermatogonial stem cell (SSCs) is unique in that it is the only cell in the adult male that can contribute genes to a subsequent generation. Permanent modification of the germ cell line may be realized if stem cells could be cultured, transfected with unique genes, and then transplanted into recipient testes. We developed a culture system that supported long-term viability of SSCs. We used a retrovirus vector (pMSCV including ${\beta}$-galactosidase) to stably transfect spermatogonia following long-term culture using the system developed. Expression of the reporter gene ${\beta}$-galactosidase controlled by the retroviral vector was stable in long-term cultured SSCs. We confirmed the retroviral-mediated ${\beta}$-galactsidase gene could be expressed in germ cells in recipient mice following SSCs transplantation.

• 한국식품저장유통학회지
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• 제5권1호
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• pp.13-21
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• 1998

Korean medicinal herb extracts(KMHE) were applied to the preservation of greenhouse produce in order to prove their effectiveness. KMHE showed remarkable antimicrobial effects against Bacillus cereus, Peudomonas syringae, and Corynebacterium xerosis causing the postharvest decay of greenhouse produce. Among KMHE the extracts of Rheum palmatum L. and Coptis chinensis Franch most obviously inhibited the growth of microorganims causing the Postharvest decay of greenhouse produce, which destroyed to undetectable levels when treated with more than 500ppm of KMHE. The activities of KMHE were stable in the wide spectrum of pH and temperature. Direct visualization of microbial cells by using both transmission electron microscope and scanning electron microscope showed microbial cell membrane the function of which was destroyed by treating with the dilute solutions of KMHE. This change of cellular membrane permeability could be identified in the experiment that O-nitrophenyl-$\beta$-D-galactopyranoside(ONPG), the artificial substrate of $\beta$-galactosidase, was hydrolyzed in the presence of KMHE, indicating that the membrane was perturbed by KMHE.

• Biomolecules & Therapeutics
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• 제6권4호
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• pp.364-370
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• 1998

Glycosidase inhibitors from urine of Bombyx mori were isolated and their inhibitory activities on glycosidases were evaluated. Six compounds were isolated by using several ion exchange columns, and their chemical structures were identified by the physicochemical and spectral data. Compound IV, V and Ⅵ were identified as 1-deoxynojirimycin, fagomine and 1,4-dideoxy-1,4-imino-D-arabinitol, respectively. Among six compounds isolated,1-deoxynojirimycin(IV) was the most potent inhibitor on $\alpha$-glucosidase and $\beta$-galactosidase of rat intestine, and its inhibitory activities for trehalase and almond $\beta$-glucosidase were relatively weak. Compound V and Ⅵl retained a little inhibitory potency toward $\alpha$-glucosidase and $\beta$-galactosidase. Compound II and III, however, have been found to have no effect on all glycosidases tested in this study.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권5호
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• pp.382-390
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• 2006

Various microorganisms were isolated from the surface waters and sediments of eutrophic lakes and reservoirs in Korea to enable an investigation of bacteria having algal lytic activities against Anabaena flos-aquae when water blooming occurs and to study enzyme profiles of algal lytic bacteria. Two bacterial strains, AFK-07 and AFK-13, were cultured, characterized and identified as Acinetobacter johnsonii and Sinorhizobium sp., respectively. The A. johnsonii AFK-07 exhibited a high level of degradatory activities against A. flos-aquae, and produced alginase, caseinase, lipase, fucodian hydrolase, and laminarinase. Moreover, many kinds of glycosidase, such as ${\beta}-galactosidase,\;{\beta}-glucosidase,\;{\beta}-glucosaminidase,\;and\; {\beta}-xylosidase$, which hydrolyzed ${\beta}-O-glycosidic$ bonds, were found in cell-free extracts of A. johnsonii AFK-07. Other glycosidases such as ${\alpha}-galactosidase,\;{\alpha}-N-Ac-galactosidase,\;{\alpha}-mannosidase,\; and\;{\alpha}-L-fucosidase$, which cleave ${\alpha}-O-glycosidic$ bonds, were not identified in AFK-07. In the Sinorhizobium sp. AFK-13, the enzymes alginase, amylase, proteinase (caseinase and gelatinase), carboxymethyl-cellulase (CMCase), laminarinase, and lipase were notable. No glycosidase was produced in the AFK-13 strain. Therefore, the enzyme system of A. johnsonii AFK-07 had a more complex mechanism in place to degrade the cyanobacteria cell walls than did the enzyme system of Sinorhizobium sp. AFK-13. The polysaccharides or the peptidoglycans of A. flos-aquae may be hydrolyzed and metabolized to a range of easily utilized monosaccharides or other low molecular weight organic substances by strain AFK-07 of. A. johnsonii, while the products of polysaccharide degradation or peptidoglycans were more likely to be utilized by Sinorhizobium sp. AFK-13. These bacterial interactions may offer an alternative effective approach to controlling the water choking effects of summer blooms affecting our lakes and reservoirs.

• Journal of Animal Science and Technology
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• 제45권2호
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• pp.211-218
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• 2003

본 연구는 양돈사료내 대두박 항영양인자인 $\alpha$-galactosides와 galatomannan의 분해를 유도하는 $\alpha$-1,6-galactosidase와 $\beta$-1,4-mannanase의 사료내 첨가가 자돈 및 육성돈의 성장과 영양소 소화율에 미치는 영향을 조사하기 위하여 실시하였다. 시험 1은 개시시 체중 10.57$\pm$0.30kg의 3원 교잡종 자돈 60두를 공시하였으며, 시험설계는 옥수수－건조유청－대두박 위주의 사료에 NRC (1998)의 영양소 요구량에 따라 처리한 대조구 (CON), 대조구 사료내 복합효소제를 0.1% 첨가한 처리구로 하였다. 사양시험기간동안, 일당증체량에 있어서는 대조구와 비교하여 EC0.1 처리구가 높은 것으로 평가되었으나 유의적인 차이는 보이지 않았다. 그러나 사료효율에 있어서는 대조구와 비교하여 EC0.1 처리구가 유의적으로 높게 평가되었다 (P<0.05). 건물과 질소 소화율에 있어서 대조구와 비교하여 처리구가 향상된 것으로 조사되었다 (P<0.05). 시험 2는 개시시 체중 22.30$\pm$0.45kg의 3원 교잡종 육성돈 36두를 공시하였으며, 시험설계는 옥수수－대두박 위주의 사료에 NRC (1998)의 영양소 요구량에 따라 처리한 대조구 (AME, adequate ME diet), 대조구 사료내 복합효소제를 0.1% 첨가한 처리구 (AME+EC0.1, Adequate ME diet+ 0.1% 복합효소제), 대조구 사료에서 대사에너지 함량을 4% 낮춘 사료에 복합효소제를 0.1% 첨가한 처리구 (LME+EC0.1, Low ME diet + 0.1% 복합효소제)로 하였다. 총 30일간의 사양시험 기간동안, 일당증체량에 있어서는 AME 처리구와 비교하여 복합효소제 처리구가 유의적인 성장율이 높은 것으로 조사되었다 (P<0.05). 건물 및 질소 소화율에 있어서는 AME 처리구와 비교하여 복합효소제 첨가구가 유의적으로 높게 평가되었다 (P<0.05). 결론적으로, 자돈 및 육성돈 사료에 복합효소제의 첨가는 성장능력 및 영양소 소화율을 향상시키는 것으로 사료된다.