• 센서학회지
• /
• 제6권6호
• /
• pp.437-444
• /
• 1997

A potentiometric L-lysine-selective sensor is described for the direct determination of lysine. The sensor system is based on a carbon dioxide gas sensing electrode and an L-lysine decarboxylase immobilized to CNBr-activated sepharose 4B. A highly selective L-lysine sensor has been prepared with immobilizing enzyme slurry put into reaction buffer solution. The optimum conditions for the measurement were evaluated by various experiments. This sensor exhibits a linear response to L-lysine concentrations from $10^{-4}M$ to $10^{-1}M$. Response time of this lysine sensor is shorter than 30secs and the immobilized enzyme slurry is stable over one year.

• Nuclear Medicine and Molecular Imaging
• /
• 제43권5호
• /
• pp.487-494
• /
• 2009

목적: 작은 크기의 재조합 단일사슬 항체는 빠른 혈중 제거율과 종양의 항체 집적율이 증가되는 등의 장점을 가지고 있다. 반면에 항체의 작은 크기는 방사성 또는 형광물질의 표지를 위한 킬레이터 결합에 중요한 아미노산 그룹의 감소를 의미하기도 한다. 본 연구에서는 단일사슬 lym-1 염기서열 C-말단에 lysine 아미노산 태그를 삽입하여 형광 물질의 직접표지 및 그 표지수율 증가를 확인하고자 하였다. 대상 및 방법: 대장균 pET-22b (+) 벡터에 재조합 된 lysine 삽입 단일사슬 lym-1유전자는 대장균 BL21 (DE3)에 형질전환하여 발현하였다. 생산된 lysine lym-1 항체는 Ni-NTA 컬럽과 분자량 컬럼을 사용해 정제하였고. 단백질 전기 영동과 western blot을 통해 확인하였다. lysine lym-1 항체에 방사성 동위원소인 I-124, I-125, I-131 과 Tc-99m를 표지하여 그 수율을 확인하였으며 유세포계측기를 사용해 형광물질인 FITC가 직접표지된 라이신 lym-1 항체의 면역반응성을 사람의 버킷 림프종 세포주인 Raji 세포주에서 면역반응성을 확인하였다. 결과 Lysine도입 단일사슬 lym-1 항체는 두 과정의 정제를 통하여 획득하였으며 그 크기는 약 48 KDa이었고, 방사성동위원소인 I-124, I-125, I-131과 Tc-99m의 표지수율은 각각 >99%, >99%, >95%, >99%로 확인되었다. 유세포계측을 통한 lysine 도입 단일사슬 lym-1항체의 면역반응성은 기존의 단일사슬 lym-1항체와 유사함을 확인하였다. 결론: 재조합 lym-1 항체에 형광 물질을 직접 표지하기 위한 lysine 아미노산의 도입은 항체의 면역반응성 감소를 최소화 시키면서 직접표지 수율을 증가시킬 수 있는 유용한 방법임을 확인하였다.

• 한국식품영양과학회지
• /
• 제10권1호
• /
• pp.77-84
• /
• 1981

The effect of sesame of L-lysine HCI and sesame supplementing a wheat flour diet on growth, liver lilpid content, and on the free amino acid levels in the plasma and liver was studied in young male rats with an initial body weight of $75{\pm}3g$. The free amino acids were analyzed by amino acid auto analyzer (JLC - 6HA, NO. 310). The results were as follows. The body weitht gain on L-lysine HCI and sesame supplemented diet was more than weight in the sesame added diet or wheat flour diet groups. Also the liver lipid contents of rats on a wheat flour diet supplemented with L-lysine HCI and sesame showed greater increases than the levels in rats on the wheat flour diets. The rate of liver lipid accumulation was depressed in rats fed L-lysine HCI supplemented wheat flour containing sesame than in rats fed soybean oil or shortening oil instead of sesame. The free phe. Tyr. Leu. Ileu. Val. Lys. levels in the plasma of rats administered the wheat flour diets supplemented with 0.25% L-lysine HCI were higher than those of rats without L-lysine HCI. The free phe. Tyr. Asp. His. Lys. contained in the liver were increased, but other free amino acids were decreased according to the L-lysine HCI amount.

• Journal of Microbiology and Biotechnology
• /
• 제24권10호
• /
• pp.1368-1376
• /
• 2014

This article focuses on the effects of glycine betaine on preventing caramelization, and increasing DCW and $\small{L}$-lysine production. The additional glycine betaine not only decreased the browning intensity (decreased 4 times), and the concentrations of 5-hydroxymethylfurfural (decreased 7.8 times) and furfural (decreased 12 times), but also increased the availability of glucose (increased 17.5%) for $\small{L}$-lysine production. The DCW and $\small{L}$-lysine production were increased by adding no more than 20 mM glycine betaine, whereas the DCW and $\small{L}$-lysine production were decreased with the reduction of pH values, although pH had a better response to prevent caramelization than did glycine betaine. For $\small{L}$-lysine production, the highest increase (40%) was observed on the media with 20 mM glycine betaine. The crucial enzymes in glycolysis and $\small{L}$-lysine biosynthesis pathway were investigated. The results indicated that additional glycine betaine increases the activity of enzymes in glycolysis, in contrast to the effect of pH. All the results indicated that glycine betaine can be used to prevent caramelization and increase the $\small{L}$-lysine production. By applying this strategy, glucose would not be have to be separated from the culture media during autoclaving so that factories can save production costs and shorten the fermentation period.

• 생명과학회지
• /
• 제17권3호통권83호
• /
• pp.444-453
• /
• 2007

유핵세포의 게놈(genome)은 단백-DNA복합체인 염색질(chromatin)의 형태로 존재하는데, 생명현상을 유지하기 위해서는 생명체 또는 세포가 처한 상황에 맞게 염색질의 구조를 변화시키는 역동적인 조절기전이 필요하다. 염색질을 구성하는 기본단위는 히스톤 8량체 (histone octamer)를 포함하는 뉴클레오좀(nucleosome)이다. 히스톤 단백에는 여러 종류의 공유결합성 수식이 일어나는데, 그 중 하나가 라이신 잔기(lysine residue)에 일어나는 메틸화이다. 최근 수년간의 연구로 여러 개의 히스톤 라이신 메틸화효소(histone lysine methyltransferase, HKMT), 이에 결합하는 염색질단백 및 메틸화와 관련된 후생유전학적 현상이 밝혀졌으며, 특히 정밀한 연구방법을 동원한 다방면의 실험을 통하여 비록 자세한 기전과 전체적인 윤곽의 규명은 미흡하더라도 라이신 메틸화가 후생유전학적 변화를 초래하는 일부 과정이 규명 되었다. 또한 여러 종류의 라이신 탈메틸화효소가 최근에 발견됨에 따라, 아세틸화, 인산화등 다른 공유결합성 수식보다는 상대 적으로 안정되더라도, 히스톤 메 틸화로 유발되는 후생유전학적 변화가 불가역성이 아님을 알게 되었다.

• 환경위생공학
• /
• 제22권2호
• /
• pp.75-84
• /
• 2007

A lysine determination method for low quantity of rice was modified from the original Dye-Binding Lysine (DBL) method used in the national standard in China [GB 4801-84, 1984]. By making use of the property that lysine does not bind to the crocein orange G dye after treated with propionic anhydride, the amount of lysine in rice samples could be determined directly by calculating the difference between the absorbances of the treated and the untreated samples. Various commercial rice samples were purchased from market and evaluated. Several methods were tested by varying both the sizes of the samples and the concentrations of the dye solutions. Results showed that when using 1.284 mM of crocein orange G dye solution and 15.5 mg of sample, the results were most reproducible. The corresponding lysine content in sample were $3.36\;{\pm}\;0.09\;mg/g$ and $3.35\;{\pm}\;0.19\;mg/g$ by traditional method and modified method, respectively. Statistically, there was no significant difference between the results (p>0.05).

• BMB Reports
• /
• 제41권11호
• /
• pp.790-795
• /
• 2008

An inducible lysine 6-dehydrogenase (Lys 6-DH), which catalyzes the oxidative deamination of the 6-amino group of L-lysine in the presence of $NAD^+$, was purified to homogeneity from Achromobacter denitrificans, yielding a homodimeric protein of 80 kDa. The enzyme was specific for the substrate L-lysine and $NAD^+$ served as a cofactor. The dimeric enzyme associated into a hexamer in the presence of 10 mM L-lysine. The $K_m$ values for L-lysine and $NAD^+$ were 5.0 and 0.09 mM, respectively. The lys 6-dh gene was cloned and overexpressed in E. coli. The open reading frame was 1,107 nucleotides long and encoded a peptide containing 368 amino acids with 39,355 Da. The recombinant enzyme was purified to homogeneity and characterized. Enzyme activities and kinetic properties of the recombinant enzyme were almost the same as those of the endogenous enzyme obtained from A. denitrificans. Crystals of the enzyme were obtained using the hanging drop method.

• Asian-Australasian Journal of Animal Sciences
• /
• 제20권8호
• /
• pp.1297-1304
• /
• 2007

We conducted a series of investigations in order to elucidate role of nutritional status in regulating GLUT expression and energy metabolism in porcine muscle. Firstly, the role of mild undernutrition in regulating muscle GLUT gene expression and function was studied in growing pigs (3 wk of age) on a high (H) or low (L) food intake (H = 2L) at $35^{\circ}C$ or $26^{\circ}C$. Low food intake selectively upregulates GLUT1 and GLUT4 gene expression; mRNA levels were elevated in longissimus dorsi (L. dorsi) and rhomboideus muscles but not in diaphragm or cardiac muscles. Our next step was to determine whether dietary lysine, a major primary limiting amino acid in diets for pigs, affects muscle GLUT4 expression. Pigs of 6 wk of age were pair-fed a control or low lysine (LL) diet. The control diet contained optimal amounts of all essential amino acids, including 1.15% lysine. The LL diet was similar but contained only 0.70% lysine. GLUT4 mRNA expression was upregulated by the LL diet in L. dorsi and rhomboideus muscles, whereas that in cardiac muscle was unaffected. GLUT4 protein abundance was also higher in rhomboideus muscle of animals on the LL diet. We conducted another investigation in order to elucidate effects of the LL diet on post-GLUT4 glucose metabolism. Activity of hexokinase was unaffected by dietary lysine levels while that of citrate synthase was higher both in L. dorsi and rhomboideus muscles of pigs fed on the LL diet. Glucose 6-phosphate content was higher in L. dorsi msucle in the LL group. Glycogen content was higher both in L. dorsi and rhomboideus muscles in the LL group. Further, we determined the effects of dietary lysine levels on accumulation of intramuscular fat (IMF) in L. dorsi muscle of finishing pigs. A low lysine diet (lysine content was 0.40%) meeting approximately 70% of the requirement of lysine was given to finishing pigs for two months. IMF contents in L. dorsi of the pigs given the low lysine diet were twice higher than those of the pigs fed on a control diet (lysine content was 0.65%). Finally, we proved that a well known effect of breadcrumbs feeding to enhance IMF of finishing pigs could be attributed to shortage of amino acids in diets including breadcrumbs.

• Asian-Australasian Journal of Animal Sciences
• /
• 제26권7호
• /
• pp.1003-1011
• /
• 2013

Two experiments were conducted to determine the effects of different dietary lysine levels on the apparent nutrient digestibility, the serum amino acid (AA) concentration, and the biochemical parameters of the precaval and portal vein blood in growing pigs. In Experiment 1, 15 noncannulated pigs received diets with different lysine densities (0.65%, 0.95%, and 1.25% lysine) for 13 d. A total collection digestion test was performed, and blood samples were collected from the precaval vein at the end of the experiment. In Experiment 2, four cannulated pigs were fed the same diets of Experiment 1. The experiment used a self-control experimental design and was divided into three periods. On d 5 of each period, at 0.5 h before feeding and hourly up to 8 h after feeding, single blood samples were collected from catheters placed in the portal vein. In Experiment 1, some serum AAs (including lysine), serum urinary nitrogen (SUN), and total protein (TP) concentrations were significantly affected by the dietary lysine levels (p<0.05). Moreover, the 0.65% lysine treatment showed a significant lower apparent digestibility of gross energy, dry matter, crude protein, and phosphorus than the other treatments (p<0.05). In Experiment 2, serum lysine, histidine, phenylalanine, threonine, valine, isoleucine (p = 0.0588), triglyceride, and SUN (p = 0.0572) concentrations were significantly affected by the dietary lysine levels (p<0.05). Additionally, almost all of the determined serum AA and total AA concentrations reached their lowest values at 0.5 h before feeding and their highest values at 2 h after feeding (p<0.05). These findings indicate that the greatest absorption of AA occurred at 2 h after feeding and that the dynamic profile of serum AA is affected by the dietary lysine levels. Moreover, when the dietary lysine content was 0.95%, the growing pigs achieved a better nutrient digestibility and serum metabolites levels.

• 한국식품과학회지
• /
• 제6권2호
• /
• pp.109-115
• /
• 1974

우리 나라 평균 식단에서 부족되고 있는 lysine을 밀쌀에 정량적으로 강화(强化)함으로써, 밀쌀 자체는 물론 우리 나라 국민의 섭취단백질을 질적으로 향상(向上)시킬 수 있는 방안(方案)을 모색코저 본 연구를 시도하였다. 따라서 밀쌀을 lysine 강화대상 곡류로 정하고, 정맥(精麥)(polished wheat) 및 밀쌀의 물리화학적 성상, 공장규모에서의 강화실험, 강화밀쌀의 저장시험 및 영양실험을 행하여 다음과 같은 결과를 얻었다. 1. 밀쌀은 호화건고(糊化乾固) 및 균열상태를 보임으로서 경도(硬度)는 정맥(精麥)보다 낮았고, 수분(水分) 및 L-lysine HCl 흡수능(吸收能)은 정맥(精麥)이 더 낮았다. 그리고 정맥(精麥) 및 밀쌀의 L-lysine HCl 흡수능(吸收能)은 침지 L-lysine HCl 용액의 농도에 거의 비례하였고 침제(浸濟) 초기에 급격히 흡수(吸收)하였다. 그리고 이들의 취반특성(炊飯特性)도 다소 차이가 있었다. 2. 공장규모(工場規模)의 L-lysine HCl 강화에 대한 일련의 실험결과, 기존 밀쌀제조공정 중의 가수공정(加水工程)에서 강화함으로써 기존 시설을 효용(效用)할 수 있었고, 강화농도가 균일한 밀쌀제품을 얻을 수 있었다. 그리고 공정 중의 L-lysine HCl 손실율은 $10{\sim}11%$이었으며, 강화밀쌀의 기호성은 보통 밀쌀과 차이가 없었다. 3. 흰쥐사육시험 결과, 보통밀쌀의 PER 1.12가 0.2% 강화수준에서 1.75로 크게 향상되었으며 0.4% 강화구에서는 1.89이었다. 4. 저장시험 결과, 실온(室溫)에서 90일, 고온과습구(高溫過濕區)($37^{\circ}C,$ >80% RH)에서 30일간 저장한 강화밀쌀 (0.2% 수준, polyethylene film packing)의 환원당 지방산도의 변화는 경미함으로서, 강화밀쌀의 저장성에 있어서는 보통 밀쌀과 같이 큰 문제점이 없을 것으로 판단되었다.