• 생명과학회지
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• 제15권3호
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• pp.415-422
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• 2005

Mycobacterium hovis BCG 균주에 transposon을 사용하여 mutagenesis를 수행함으로써 mutant library를 제조하였다. 이 mutant library를 screening하여 항결핵제인 PA-824에 내성을 갖는 mutant들을 얻었고, M. bovis wild type에서는 정상적으로 생성되는 coenzyme $F_{420}$이 대부분의 이들 mutant들에서는 생성되지 않는다는 것을 알게 되었다. 세포 추출액을 HPLC로 분석해본 결과, 그 중에서 한 mutant는 $F_{420}$은 생성하지 않으나 그 전구물질인 F0는 생성하고 있음이 밝혀졌다. 따라서 이 mutant 에서는 $F_{420}$생합성 회로의 마지막 단계가 차단되어있음을 알 수 있다. 이 mutant를 inverse PCR을 통해 분석해본 결과, transposon이 Rv2435c유전자에 삽입되어있는 것을 확인할 수 있었다. Rv2435c유전자는 세포막에 결합되어있는 80.3 kDa의 단백질을 암호화하는 것으로 추정되고, 이 단백질의 N-말단은 periplasm에 존재하고 C-말단은 원형질에 존재하는 것으로 추정되고 있다. 원형질에 존재하는 C-말단은 원핵생물과 진핵생물들의 adenylyl cyclase들과 높은 유사성을 나타낸다. Adenylyl cyclase는 ATP로부터 cAMP를 생합성하는 효소이다 M. tuberculosis나 M. bovis의 genome에는 class III adenylyl cyclase를 암호화하는 것으로 추정되는 유전자가 모두 15개나 존재한다 특히 이들 중에서 Rv1625c 와 Rv2435c는 포유류의 adenylyl cyclase들과 높은 유사성을 가지는 것으로 알려져 있다 이 Rv2435c 단백질이 진정한 adenylyl cyclase인지를 확인하기 위하여 우리는 이 단백질 중에서 원형질에 존재하는 부분을 말단에 6개의 histidine을 첨부한 채로 대장균에서 발현시켰다. 대장균에서 이 단백질이 생성되는 것은 histidine이 첨부된 단백질을 Ni-NTA resin을 사용하여 대장균으로부터 분리함으로써 확인하였다. 그러나 이 단백질이 대장균에서 cya mutation을 complementation하지 못하였고, 따라서 이 단백질이 adenylyl cyclase 활성을 갖지 않음을 알 수 있었다. 자외선이나 hydroxylamine을 사용한 mutagenesis 또는 Rv2435c와 Rv1625c간의 토sion단백질을 만들어서, 이 단백질이 adenylyl cyclae로서의 활성을 획득하도록 하는 모든 시도는 실패하였다. 따라서 Rv2435c단백질이, F0가 $F_{420}$으로 변환되는 데에 영향을 미치는 방법이 cAMP를 생성함으로써가 아니라 다른 방법으로 영향을 미치고 있다는 것을 알 수 있었다.

• 기술사
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• 제18권2호
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• pp.16-22
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• 1985

This study was carried out to evaluate the quality of soybean sprouts grown under three different sources of light. The soybean was germinated under light through blue (251 ux), red (751 ux) and white (50 to 200 lux) polyethylene films at 25$^{\circ}C$ for 10hr./day. Vitamin C, chlorophyll, cellulose and total protein contents were determined and texture was evaluated by tasting soybean sprouts soup. protein pattern in the soybean sprouts were investigated using polyacrylamide gel electrophoresis. Vitamin C, chlorophyll and cellulose contents were increased by white light intensity. The texture of the sprouts grown under white light (50 lux, 100 lux) was better than darkness, but fought under 200 lux. Vitamin C contents of soybean sprouts grown under various sources of light (in order of light : blue > white > red) were higher than theses of ones grown in the darkness. Biosynthesis of chlorophyll was not correlated to Vitamin C content. Total protein contents of cotyledon was not changed significantly under light irradiation. But the soybean sprouts grown under different quality of light, hypocotyl was higher than those grown darkness. (37% and 20% higher for blue light and white light) Densitometric tracing of disc polyacrylamide gel electrophoretic patterns showed that protein of hypocotyl under white light had more high molecular weight protein.

• 한국미생물·생명공학회지
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• 제17권6호
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• pp.556-562
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• 1989

The bioconversion of mandarin orange peel press liquor to single cell protein (SCP) by two yeast strains, F-60, and C-7, which were isolated from mandarin orange peel was carried out and compared with that of using Candida utilis IFO 0598. Experiments were directed toward the high yield of biomass and high protein in cultures of the strains mentioned above. Candida utilis IFO 0598, F-60 and C-7 strains were cultivated at 3$0^{\circ}C$, pH 5.2 for 3 days in shaking flasks. The effects of some nutrients on cell growth were studied. Cell mass and protein content per cell mass were increased by addition of urea 1%, KH$_2$PO$_4$ 0.1% and MgSO$_4$ㆍ7$H_2O$ 0.05%, When the F-60 strain cultured under the optimal conditions, cell mass, growth yield and protein content were 41.2g/l, 53.9%, 59.7%, respectively. Cell mass was also increased up to 15% by modifying the fermentation condition on the bench type 20l jar fermentor. Crude fat content (10.3%) of dried C-7 cell was higher than those of C. utilis and F-60, 4.9% and 5.6% respectively. Total protein content of the F-60 strain was 59.7% per dry weight. And we compared their amino acid compositions with that of FAO provisional pattern. In the case of the F-60 strains, amino acid contents such as lysine, leucine and isoleucine were much higher than those of methionine, cystine and tryptophan.

• KSBB Journal
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• 제8권2호
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• pp.143-149
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• 1993

Activated carbons were used to examine their performance for the separation of undesirable colored materials from culture broth containing hyaluronic acid. Six local samples and a NORIT ROX 08 were tested, whereas the latter was mainly studied under batch and continuous modes. The optimal wavelength for the detection of colored materials was 330nm. The optimal choice of NORIT ROX 08 provided 30% colored residuals with 96% hyaluronic acid recovery of original broth in batch experiments. The nonlinear adsorption behavior of protein and colored materials with activated carbon (C) was correlated by a Langmuir equation to give 18C/24+C and 500C/892+C for protein and colored materials, respectively. It appears that colored materials were composed of 78% protein and 22% glucose residuals on the basis of clearance results. A microscopic study using a scanning electron microscope suggests that regeneration of used activated carbon with 0.1N NaOH and hot water was not satisfactory. The present study proposes that the continuous monitoring of colored materials during purification can be accomplished by Installation of a UV monitor commonly used for continuous detection of protein during the process, as resulted from the significant correlation of color (A330)=0.353protein(mg/ml)+0.1(R=99.7%).

• 한국조리학회지
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• 제24권1호
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• pp.24-30
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• 2018

This study investigated the quality of characteristics of high-protein RTD coffee using domestic and imported skim milk powder with different heat treatment. Skim milk powders (A, B) had high-heat treatment, C had medium-heat, and D and E had low-heat treatment. The transmittance of A and B were higher than that of C, and that of C were higher than that of D and E (p<0.05). The precipitate attached on bottom of container of RTD coffee using A and B were 2.993~3.053% and higher than those (0.753~0.803%) of RTD coffee using C, D and E (p<0.05), but there was no difference between those of RTD coffee using C, D and E (p<0.05). The centrifuged precipitate of RTD coffee using A and B were 1.987~2.040% and higher than those (0.820~0.830%) of RTD coffee using C, D and E (p<0.05), but there was no difference between those of RTD coffee using C, D and E (p<0.05). The proximate composition of precipitate attached on bottom of container of RTD coffee using A, which showed the highest amount of precipitate, showed 65.7% of carbohydrate, 19.0% of protein, 4.8% of fat and 4.8% of ash in dry basis, that of RTD coffee being 72.7%, 15.1%, 7.9% and 4.3% in dry basis respectively. Protein and fat content of precipitate were lower and protein and ash content were higher than those of RTD coffee. But seeing that the most increased portion was protein, precipitation of RTD coffee appears to be attributed to heat-denatured proteins.

• 한국식품과학회지
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• 제26권2호
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• pp.103-109
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• 1994

응력완화 현상을 측정하여 생선단백질 겔의 선형적 점탄성과 열처리 공정에 따른 물성의 변화를 수식적 모델로 분석하였다. 생선단백질 겔은 정변형도 $0.105{\sim}0.693$, 압축속도 $50{\sim}250\;mm/min$의 범위에서 선형적 점탄성을 나타내었으며, generalized Maxwell 모형에 의해 분석한 결과, 압축변형도과 압축속도가 증가함에 따라 가열에 의해 형성된 내부조직 중 탄성 성분의 증가와 점성성분의 감소현상을 보였다. $4^{\circ}C$와 $40^{\circ}C$에서 전처리하여 $90^{\circ}C$에서 제조한 겔은 전처리 없이 $90^{\circ}C$에서 직접 열처리한 겔보다 탄성율(E) 및 평형탄성율$(E_e)$이 높았으나, 점성성분$({\eta})$은 적용된 모델에 따라서 그 값의 차이가 나타났다. 따라서 식품의 물성을 측정하는데 있어서 두 수학적 모델의 접근방법 및 정확도에 대하여 설명하였다.

• KSBB Journal
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• 제28권5호
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• pp.310-314
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• 2013

Antifreeze peptide from Myoxocephalus octodecemspinosus was overexpressed and purified in Escherichia coli. Green fluorescence protein-AFP chimera was constructed by integrating gfp and afp genes. Produced GFP-AFP chimera protein was purified using polyhistidine tag which was inserted at C-terminus. By addition of GFP-AFP chimera protein, freezing point of elution buffer was decreased from $-13^{\circ}C$ to $-20^{\circ}C$. This result suggested that GFP-AFP chimera can be considered as a potential candidate of novel inhibitor for gas hydrates.

• Food Science and Biotechnology
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• 제14권1호
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• pp.11-15
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• 2005

Dynamic shear moduli of isolated soy protein solutions upon heating were measured to monitor gelation. Onsets of gelation coincide with onset temperatures of denaturation in glycinin and ${\beta}$-conglycinin solutions, whereas in isolated soy proteins, onset of gelation was above denaturation temperature of ${\beta}$-conglycinin with storage modulus increasing in two steps. The first increase in storage modulus of isolated soy proteins occurred at about $78.5^{\circ}C$, while the second increase started at about $93^{\circ}C$. Gel properties of soy protein gels having different proportions of glycinin and ${\beta}$-conglycinin were measured by compression-decompression test. ${\beta}$-conglycinin was responsible for gel elasticity. Glycinin significantly increased hardness, toughness, and fracturability of gels at high heating temperature near $100^{\circ}C$. Results reveal texture of soy protein gels can be controlled by regulating ratio of glycinin to ${\beta}$-conglycinin and heating temperature.

• 대한화학회지
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• 제21권3호
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• pp.215-218
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• 1977

장광품종의 밀 gliadin 단백질을 정제하여 주 gliadin 단백질을 얻은 다음 이의 아미노산조성과 분자량 그리고 N-말단 및 C-말단 아미노산을 조사하여 본 결과 1. 주 gliadin 단백질의 아미노산 조성은 glutamic acid의 함량이 가장 많았다. 2. 주 gliadin 단백질의 분자량은 60,200 ${\pm}$200이었다. 3. 주 gliadin 단백질의 N-말단 아미노산은 phenylalanine이었고, C-말단 아미노산은 methionine이었다.

• Toxicological Research
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• 제12권2호
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• pp.155-161
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• 1996

The enzymatic properties for NADPH-P450 reductase domain of a fusion protein between human cytochrome P450 1A1 and rat NADPH-P450 reductase expressed in Escherichia coli were investigated. The fusion plasmid pCW/1A1OR-expressed E. coli membrane showed high NADPH-cytochrome c reductase activity ($830.1\pm 85.8 nmol\cdot min^{-1}\cdot mg protein^{-1}$), while pCW control vector and P 450 1A1 expression vector pCW/1A1 showed relatively quite low activity ($4.35\pm 0.49, 3.27\pm 0.50 nmol\cdot min^{-1}\cdot mg protein^{-1}$, respectively). The kinetic curves for NADPH-cytochrome c reductase followed typical Michaelis-Menten kinetics. The $K_{max}$ and $V_{max}$ for NADPH-dependent reductase activity were $8.24\pm 2.61\mu $and $817.9\pm 60.8 nmol\cdot min^{-1}\cdot mg protein^{-1}$, respectively, whereas those for cytochrome c-dependent reductase activity were $19.97\pm 2.86\mu M$ and $1303.5\pm 67.1 nmol\cdot min^{-1}\cdot mg protein^{-1}$. The reductase activities were also compared with those of rat, porcine and human liver microsomes. The activity of pCW/ 1A1OR-expressed E. coli membrane was 15.2-fold higher than that of rat liver microsome. Treatment with benzo(a)pyrene, 7-ethoxyresorufin and $\alpha$-naphthofiavone which are known as specific substrates or inhibitor for human P450 1A1 increased NADPH-cytochrome c reductase activity of fusion protein in E. coli membrane dose-dependently. These results demonstrate that the membrane topology of fused enzyme may be important for activity of its NADPH-P450 reductase domain.