• 한국작물학회지
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• 제27권2호
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• pp.147-154
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• 1982

식물원형질체를 이용하여 체세포잡종을 육성하기 기초자료를 얻고자 식물의 엽육부위, Callus에서 효과적인 원형질체의 분리, 적합한 배양조건 및 세포융합에 관한 조건을 조사한 결과를 요약하면 다음과 같다. 1. 배양 융합조건에 적합한 식물원형질체의 분리 방법은 여러 종류의 효소농도에 따라 다르지만 담배 및 완두엽육 원형질체분리에는 Macerozyme 0.5%, Cellulase 2.0%, KDS 0.5%, Mannitol 0.7M의 효소용액에 처리시 활성 있는 원형질체를 얻을 수 있었다. 2. 배양중인 담배 및 대두의 Callus로부터 원형질체분리는 배양기간에 따라 세포활성도가 떨어지거나 쉽게 파괴되는 현상을 보여 양호한 원형질체를 얻을 수가 없었다. 3. 원형질체의 배양은 배양배지에 Plating할 경우 세포농도에 따라 차이가 있었으며 1.0$\times$$10^4$/ml이상이어야 분열이 가능하였고 배양조건은 150Lux 12시간 광암처리후가 세포분열 및 증식이 가장 왕성하였다. 4. 세포융합은 PEG농도가 중요하며 PEG 1500 0.33M에서 CaCl_2농도가 높을수록 융합율이 증가되는 경향이었다.

• 한국지반공학회논문집
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• 제39권8호
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• pp.7-16
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• 2023

본 연구는 매립지 침출수 내 황산염 농도를 저감하기 위해 친환경 지반개량 공법인 Enzyme Induced Carbonate Precipitation(EICP) 공법을 활용하였다. 황산염의 화학적 침전을 유도하기 위해 충분한 탄산칼슘을 생성함과 동시에 여분의 칼슘 이온을 남길 수 있는 최적의 EICP 혼합비가 계산되었다. 최적 혼합비로 처리된 사질토 시편에서 황산염 침전이 전단 강성도에 미치는 영향을 확인하고자 전단파 속도를 측정하였고 전단파 속도 측정은 EICP 반응 및 황산염 반응 시간동안 수행되었다. 실험 결과, 생성된 침전물에 따른 전단 강성도의 발달을 확인하였고 주사전자현미경(SEM)으로 침전물의 유형 및 패턴을 시각적으로 관찰하였다. 고순도 우레아제의 대체제로서 백태가루를 효소로 사용한 EICP 용액의 경우 고순도 EICP 용액과 동일한 탄산칼슘 생성 효율에서 보다 낮은 황산염 제거 효율을 보였는데 이는 백태가루에 포함된 불순물이 석고의 침전을 방해하기 때문이다.

• Asian-Australasian Journal of Animal Sciences
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• 제16권10호
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• pp.1501-1509
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• 2003

The simultaneous addition of xylanase (5,600 EXU/kg) and phytase (500 FTU/kg) feed enzymes to wheat-based broiler diets was investigated. Starter, grower and finisher diets, with three tiers of nutrient specifications, were fed to 1,440 broiler chicks kept on deep litter from 1-42 days post-hatch, without and with xylanase plus phytase, to determine the effects of diet type and enzyme supplementation on growth performance. The nutrient specifications of type A diets were standard; energy density and protein/amino acid levels were reduced on a least-cost basis to formulate type B diets and further reduced to type C diets. Phosphorus (P) and calcium (Ca) levels were adjusted in supplemented diets. From 1-42 days post-hatch, diet type significantly influenced growth performance. Birds on type C diets had lower growth rates (2,429 vs. 2,631 g/bird; p<0.001), higher feed intakes (4,753 vs. 4,534 g/bird; p<0.005) and less efficient feed conversion (1.96 vs. 1.72; p<0.001) than birds offered type A diets. Enzyme supplementation increased growth rates by 3.2% (2,580 vs. 2,501 g/bird; p<0.005) and improved feed efficiency by 2.7% (1.80 vs. 1.85; p<0.05) over the entire feeding period. There were no interactions between diet type and enzyme supplementation. At 21 days, 5 out of 30 birds per pen were transferred to cages to ascertain treatment effect on apparent metabolisable energy (AME) and nitrogen (N) retention. Xylanase plus phytase enhanced AME (13.48 to 13.91 MJ/kg DM; p<0.001) and N retention (56.3 to 59.7%; p<0.005). Carcass and breast weights of the caged birds were determined following commercial processing. Diet type significantly influenced breast weight, carcass weight and yield. Birds offered Type A diets, in comparison to Type C diets, supported heavier breast (467 vs. 424 g; p<0.001) and carcass weights (1,868 vs. 1,699 g; p<0.001) with superior carcass yields (71.8 vs. 70.6%; p<0.005). Enzyme addition increased carcass weight by 3.9% (1,752 vs. 1,821 g; p<0.005) and breast weight by 5.8% (431 vs. 456 g; p<0.01) without influencing yields. Feed ingredient costs per kg live weight gain and per kg carcass weight indicated that enzyme addition was economically feasible, where supplementation of Type A diets generated the most effective results. Importantly, soluble and total non-starch polysaccharide and phytate contents of the wheat used were typical by local standards. This study confirms the potential of supplementing wheat-based broiler diets with xylanase plus phytase but further investigations are required to define the most appropriate inclusion rates and dietary nutrient specifications in this context.

• 한국식품영양과학회지
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• 제31권1호
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• pp.92-97
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• 2002

갈화와 갈근이 알코올성 간손상 흰쥐의 알코올 대사효소계에 미치는 영항을 구명하기 위하여 알코올을 투여한 흰쥐에게 갈화와 갈근 열수추출물을 2수준으로 나누어 5주간 급여한 후 그 효과를 관찰하였다. 체중증가량, 식이섭취량과 식이효율은 알코올 투여시 감소하였으며 갈화와 갈근추출물 급여로 회복되는 경향을 보였다. 장기 무게에 미치는 칡추출물의 영향은 알코올 투여로 증가된 간, 뇌와 심장 무게를 감소시켰는데 갈화가 갈근에 비하여 뇌와 심장 무게의 감소효과가 큰 것으로 나타났다. 알코올 대사효소 활성 변화에서 알코올 투여시 유의적으로 증가된 ADH와 MEOS 활성은 칡추출물 급여시 감소된 반면 AlDH 활성은 대조군에서 유의적으로 감소되었는데 칡추출물 급여시 그 활성이 증가되었다. MAO 활성과 과산화수소 함량도 알코올 투여시 증가되었으며 갈화와 갈근 열수추출물 급여시 유의적으로 감소되었다. 이상의 결과에서 갈화와 갈근 열수추출물 급여는 알코올 중독된 흰쥐의 알코올 대사계 효소 활성 변화에 영향을 미쳐 알코올성 간손상 완화에 효과적일 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제10권4호
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• pp.335-356
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• 1997

Injection of bovine growth hormone (bGH) to lactating dairy cows increases milk yield and yields of milk components including fat. It is generally believed that most of the anabolic effects derived from bGH in animal tissues are primarily mediated by IGF-1. IGF-1 is a strong anabolic peptide in the plasma of animals and exerts mitogenic and metabolic effects on target cells. Contrary to most protein hormones, the majority of IGF-1 in circulation is bound to the binding proteins (IGFBPs) which are known to be responsible for modifying the biological actions of IGF-1, thus making determinations of IGF-1 actions more difficult. On the other hand, fat is a major milk component and the greatest energy source in milk. Currently, the fat content of milk is one of the major criteria used in determining milk prices. It has been known that flavor and texture of dairy products are mainly affected by milk fat and its composition. Acetyl-CoA carboxylase (ACC) is the rate limiting enzyme which catalyzes the conversion of acetyl-CoA to malonyl-CoA for fatty acid synthesis in 1ipogenic tissues of animals including bovine lactating mammary glands. In addition to the short-tenn hormonal regulation of ACC by changes in the catalytic efficiency per enzyme molecule brought about by phosphorylation and dephosphorylation of the enzyme, the long-term hormonal regulation of ACC by changes in the number of enzyme molecules plays an essential role in control of ACC and lipogenesis. Insulin, at supraphysiological concentrations, binds to IGF-1 receptors, thereby mimicking the biological effects of IGF-1. The receptors for insulin and IGF-1 share structural and functional homology. Furthermore, epidermal growth factor increased ACC activity in rat hepatocytes and adipocytes. Therefore, it can be assumed that IGF-1 mediating bGH action may increase milk fat production by stimulation ACC with phosphorylation (short term) and/or increasing amounts of the enzyme proteins (long term). Consequently, the main purpose of this paper is to give the readers not only the galactopoietic effects of bGH, but also the insight of bGH action with regard to stimulating milk fat synthesis from the whole body to the molecular levels.

• 생명과학회지
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• 제23권12호
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• pp.1471-1476
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• 2013

Duolite A568 담체에 흡착으로 ${\beta}$-galactosidase를 고정화시켰다. 효소의 흡착되는 현상은 Freundlich 흡착 등온식을 잘 따랐다. 흡착에 관여된 파라미터인 k와 n은 각각 14.62와 1.744를 얻었다. 자유 효소와 고정화 효소의 속도식에 관여된 매개변수를 구하기 위해 초기 속도법을 실시하였다. Michaelis-Menten 상수($K_m$)는 고정화 효소가 120 mM이고 자유 효소가 79 mM 이었다. 재순환 충진층 반응기에서 갈락토스의 농도를 변화시키면서 경쟁적 저해식에 대한 영향을 조사하였다. 갈락토스에 의한 경쟁적 저해식에 대한 모델이 실험 결과와 잘 일치하였으며 $V_m$, $K_m$ 그리고 $K_I$ 값은 각각 46.3 $mmolmin^{-1}mg^{-1}$, 120 mM and 24.4 mM 이었다. 연속 충진층 반응기에서 락토스 용액의 유량을 증가시킬 때 서로 다른 락토스 농도에서 락토스의 전환율이 감소하였다. 장기 연속 조업을 통해 고정화 효소의 안정성을 평가하기 위해 11일 동안 연속적으로 반응기 운전을 실시하였다. 고정화 효소의 잔류하는 활성은 63%로 유지되었고 효소의 반감기는 15일로 밝혀졌다.

• 동아시아식생활학회지
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• 제21권5호
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• pp.731-738
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• 2011

콜레스테롤의 신속하고 정확한 새로운 분석방법을 모색하기 위하여 본 연구에서는 전기적 전도성이 우수한 MWCNT를 이용하여 전극을 제작하였고, 여러 가지 효소고정화 방법을 통해 전기화학적 감응도 분석을 실시하였다. MWCNT의 전도성을 향상시키기 위해 아민기를 도입한 MWCNT-$NH_2$를 제조하였고, MWCNT-$NH_2$/GCE에 PB를 점착하여 작업전극을 제조하였다. 제조한 작업전극은 0.5~500 ${\mu}M$ $H_2O_2$ 농도 범위에서 농도가 증가함에 따라 전류가 비례적으로 증가하였고, 검출한계는 0.1 ${\mu}M$로 나타나 전극이 높은 감도를 가지고 있음을 확인하였다. 또한 콜레스테롤 검출을 위해 적합한 효소 반응기를 제작하기 위해 담체인 aminopropyl glass beads, CNBr-activated sepharose, Na-alginate, toyopearl beads에 cholesterol oxidase를 고정화시켜 바이오센서의 콜레스테롤 표준용액에 대한 감응도를 측정한 결과, aminopropyl glass beads과 CNBr-activated sepharose는 1~100 ${\mu}M$ 범위에서 선형관계를 보였으며, Na-alginate는 5~50 ${\mu}M$의 범위에서, toyopearl beads는 1~50 ${\mu}M$ 범위에서 선형관계를 나타내었다. 검출한계는 제작된 효소반응기 모두 1 ${\mu}M$로 나타나 콜레스테롤에 대한 높은 검출력을 보여주었으나, 특히 CNBr-activated sepharose와 Na-alginate를 이용한 효소반응기가 높은 coupling efficiency와 감응도를 보여 콜레스테롤 검출을 위한 본 바이오 센서 시스템에 적합한 것으로 나타났다.

• Asian-Australasian Journal of Animal Sciences
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• 제23권6호
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• pp.825-832
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• 2010

The United Nations Convention on Biological Diversity defines biotechnology as "Any technological application that uses biological systems, dead organisms, or derivatives thereof, to make or modify products or processes for specific use" Biotechnology has made tremendous contributions to improve production efficiency of agriculture during the last century. This article reviews successful examples of application of bio-fermentation in improving swine nutrition efficiency mainly based on the authors'z own research experience. Production of feed grade supplemental amino acids by bio-fermentation allowed nutritionists to formulate accurate feed for optimal lean growth and reduced nitrogen excretion. Recent issues with high feed grain prices caused potential feed quality problems. Bio-fermentation allowed nutritionists to use exogenous supplemental enzymes such as phytase and NSPases in swine diets, thereby improving nutrient utilization and reducing nutrient excretion to the environment. Yeast metabolites are also produced by bio-fermentation and have been repeatedly shown to improve milk production of sows during early lactation even though actual mechanisms are still to be investigated. Bio-fermentation technology also allowed nutritionists to prepare vegetable protein sources with large protein molecules and anti-nutritional factors suitable for feeding newly weaned piglets, as selected microorganisms significantly reduce specific anti-nutritional factors and size of peptides. Preparations of vegetable protein sources suitable for newly weaned pigs will greatly contribute to swine nutrition by providing efficient alternatives to the use of animal protein sources that are often expensive and somewhat against societal preference. Considering the few examples listed above, biotechnology has closely influenced improvement of production efficiency in the swine industry. As we have limited resources to produce meat to satisfy ever-increasing global demands, extensive adaptation of biotechnology to enhance production efficiency should be continued. However, at the same time, wise and careful application of bio-technology should be considered to ensure production of safe food and to meet the expectations of our society.

• Journal of Microbiology and Biotechnology
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• 제18권2호
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• pp.287-294
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• 2008

Three amino acid residues (His119, Glu164, and Glu338) in the active site of Thermus caldophilus GK24 ${\beta}$-glycosidase (Tca ${\beta}$-glycosidase), a family 1 glycosyl hydrolase, were mutated by site-directed mutagenesis. To verify the key catalytic residues, Glu164 and Glu338 were changed to Gly and Gln, respectively. The E164G mutation resulted in drastic reductions of both ${\beta}$-galactosidase and ${\beta}$-glucosidase activities, and the E338Q mutation caused complete loss of activity, confirming that the two residues are essential for the reaction process of glycosidic linkage hydrolysis. To investigate the role of His119 in substrate binding and enzyme activity, the residue was substituted with Gly. The H119G mutant showed 53-fold reduced activity on 5mM p-nitrophenyl ${\beta}$-D-galactopyranoside, when compared with the wild type; however, both the wild-type and mutant enzymes showed similar activity on 5mM p-nitrophenyl ${\beta}$-D-glucopyranoside at $75^{\circ}C$. Kinetic analysis with p-nitrophenyl ${\beta}$-D-galactopyranoside revealed that the $k_{cat}$ value of the H119G mutant was 76.3-fold lower than that of the wild type, but the $K_m$ of the mutant was 15.3-fold higher than that of the wild type owing to the much lower affinity of the mutant. Thus, the catalytic efficiency $(k_{cat}/K_m)$ of the mutant decreased to 0.08% to that of the wild type. The $k_{cat}$ value of the H119G mutant for p-nitrophenyl ${\beta}$-D-glucopyranoside was 5.l-fold higher than that of the wild type, but the catalytic efficiency of the mutant was 2.5% of that of the wild type. The H119G mutation gave rise to changes in optima pH (from 5.5-6.5 to 5.5) and temperature (from $90^{\circ}C\;to\;80-85^{\circ}C$). This difference of temperature optima originated in the decrease of H119G's thermostability. These results indicate that His119 is a crucial residue in ${\beta}$-galactosidase and ${\beta}$-glucosidase activities and also influences the enzyme's substrate binding affinity and thermostability.

• Asian-Australasian Journal of Animal Sciences
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• 제16권8호
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• pp.1158-1164
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• 2003

In two feeding experiments male and mixed-sex broiler chicks were offered diets based on sorghum and a wheatsorghum blend with two tiers of nutrient specifications, without and with microbial phytase (600 and 800 FTU/kg), from 7-25 and 1-42 days post-hatch, respectively. The nutrient specifications for protein, amino acids, energy density and phosphorus (P) of standard diets were reduced to formulate the modified diets on a least-cost basis. Calculated differences in nutrient specifications between standard and modified diets ranged from 14.3 to 17.1 g/kg crude protein, 0.24 to 0.40 MJ/kg apparent metabolisable energy (AME) and 1.06 to 1.20 g/kg available P. In both experiments, reduced nutrient specifications had a negative impact on growth rates and feed efficiency and phytase supplementation had a positive influence on growth performance and protein efficiency ratios (PER). Phytase addition to the less expensive, modified diets either partially or entirely compensated for reduced growth performance and, consequently, feed costs per kg of live weight gain were reduced. In Experiment 1, phytase increased (p<0.001) nitrogen-corrected AME (AMEn) from 15.39 to 15.89 MJ/kg dry matter. For nitrogen (N) retention there was an interaction (p<0.05) between diet type and phytase as the effects of phytase on N retention were more pronounced in the modified diets, with an increase from 0.512 to 0.561. These results demonstrate the positive effects of phytase on protein and energy utilisation, in addition to its established liberation of phytate-bound P and illustrate the feasibility of assigning nutrient replacement values to the feed enzyme for consideration in least-cost ration formulations. Further work is, however, required to define the most appropriate reductions in nutrient specifications in association with phytase supplementation.