• Title/Summary/Keyword: L-Proline

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Purification and Characterization of Two Extracellular Glucoamylase Isozymes from Lipomyces kononenkoae CBS 5608 Mutant

  • Chun, Soon-Bai;Bai, Suk;Im, Suhn-Young;Choi, Won-Ki;Lee, Jin-Jong
    • BMB Reports
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    • v.28 no.5
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    • pp.375-381
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    • 1995
  • Two forms of glucoamylase (GI and GII) from starch-grown Lipomyces kononenkoae CBS 5608 mutant were purified to apparent homogeneity by means of ultrafiltration, Sephacryl S-200 gel filtration and DEAE Sephadex A-50 chromatography. The apparent molecular weight was calculated as ca. 150 kDa for GI and ca. 128 kDa for GII, respectively. Both enzymes were glycoproteins with isoelectric points of 5.6 (GI) and 5.4 (GII). They had a pH optimun of 4.5 and were stable from pH 5 to 8. The temperature optimum for both enzymes was $60^{\circ}C$, but they were rapidly inactivated above $70^{\circ}C$. The $K_m$ values toward starch were estimated to be 6.57 mg per ml for GI and 4.52 mg per ml for GII, and the $V_{max}$ values were 16.28 ${\mu}M$ per mg for GI and 32.25 ${\mu}M$ per mg for GII, respectively. The $K_m$ and $V_{max}$ values of GII for ${\alpha}-$ or ${\beta}-cyclodextrin$ were estimated to be 0.15 mg per ml and 2.0 mg per ml, respectively ($K_m$) and 1.02 ${\mu}M$ per mg or 1.02 ${\mu}M$ per mg, respectively ($V_{max}$). Neither enzyme exhibited pullulanase activity but they released only glucose from starch or cyclodextrin. Amino acid analysis indicated that both glucoamylases were enriched in proline and acid amino acids. Glucoamylase GII strongly cross-reacted with a monoclonal antibody raised against GI enzymes, and the two enzymes shared very similar amino acid composition. Western blot analysis indicated that L. kononenkoae CBS 5608 mutant produced two forms of glucoamylase on starch, and that synthesis of them was subject to glucose repression.

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Antioxidant and ACE Inhibiting Activities of the Rockfish Sebastes hubbsi Skin Gelatin Hydrolysates Produced by Sequential Two-step Enzymatic Hydrolysis

  • Kim, Hyung-Jun;Park, Kwon-Hyun;Shin, Jun-Ho;Lee, Ji-Sun;Heu, Min-Soo;Lee, Dong-Ho;Kim, Jin-Soo
    • Fisheries and Aquatic Sciences
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    • v.14 no.1
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    • pp.1-10
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    • 2011
  • This study was conducted to obtain hydrolysates with potent antioxidative activity from rockfish skin gelatin. Gelatin was extracted under high temperature/high pressure using a two-step enzymatic hydrolysis with commercial enzymes such as Alcalase, Flavourzyme, Neutrase, and Protamex. The second rockfish-skin gelatin hydrolysate (SRSGH) was prepared by further incubating the first gelatin hydrolysate (FRSGH), which had been hydrolyzed with Alcalase for 1-h (FRSGH-A1), with Flavourzyme for 2-h (SRSGH-F2). The second gelatin hydrolysate showed higher antioxidative activity of 3.72 as measured by a Metrohm Rancimat and superior angiotensin I-converting enzyme (ACE) inhibiting activity of 0.82 mg/mL. Compared with the gelatin, the relative proportion in SRSGH-F2 was markedly decreased in the 100-kDa peak, whereas it was increased in that less than 100-kDa. The amino acid composition of SRSGH-F2 was rich in glycine (25.9%), proline (10.8%), alanine (9.1%), and glutamic acid (9.1%). In contrast, it was poor in cystine (not detected), methionine (1.6%), tyrosine (0.4%), hydroxylysine (0.9%), and histidine (0.9%). In recent years, demand for natural functional foods has been increasing, and SRSGH-F2 can be used as a functional food ingredient in the food industries. However, further detailed studies on SRSGH-F2 with regard to its antioxidant activity in vivo and the various antioxidant mechanisms are needed.

A Study on Korean Green Tea(II) -Analysis of free Amino Acid and Mineral- (한국산(韓國産) 녹다(緣茶)에 대(對)한 연구(硏究) (제 II 보) -Free amino Acid와 무기성분(無機成分)에 대(對)하여-)

  • Jung, Jai-Kie;Yu, Choon-Hie;Chung, Tai-Young;La, Sang-Moo
    • Journal of Nutrition and Health
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    • v.6 no.3
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    • pp.17-20
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    • 1973
  • Quantitative analysis was achieved by gas-liquid chromatographic method (GLC) with a single column system of OV-17 for 16 of free amino acids in Korean green tea and the contents of mineral in it was determined by atomic absorption flame emission. The results are summarized as follows: 1) Korean green tea contained Mn 1% or over out of the total ash content and $0.05{\sim}0.20%$ in the water extraction, as the major mineral. 2) Ba, Cr, Ni, Pb and V were analyzed also by small quantities relatively and Co, Tin and Ywere not detected in the water extraction. 3) GLC indicated the presence of 16 components in free ammo acids. 4) The quantities of free amino acids in Korean green tea were determined $2.96{\sim}6.61%$ Alanine, $1.01{\sim}l.89mg%$ Glycine, $2.07{\sim}7.81mg%$ Valine, $1.27{\sim}8.76mg%$ Leucine and Isoleucine, $94.31{\sim}316.27mg%$ Threonine, $9.10{\sim}39.91mg%$ Serine, $2.18{\sim}36.76mg%$ Hydroxyproline, $2.72{\sim}5.90mg%$ Proline, $39.64{\sim}70.02mg%$ Aspartic Acid, $25.93{\sim}101.28mg%$ Glutamic Acid and Lysine, $8.32{\sim}18.30mg%$ Phenylalanine and Tyrosine in trace amount. 5) The total free amino acid contents in Korean green tea ranged from 207.24mg% to 516.06mg% and Moo-Deoung tea contained outstandingly high, 516 mg% or over.

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Isolation of Coat Protein Gene from Cucumber Mosaic Virus and Its Introduction into Tobacco (오이 모자이크 바이러스 외피 단백질 유전자 분리 및 담배로의 형질전환)

  • 손성한;김경환;김영태;박종석;김주곤;이광웅;황영수
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.3
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    • pp.149-155
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    • 1995
  • The coat protein (CP) gene was cloned from RNA genome of the Cucumber Mosaic Virus strain ABI (CMV-ABI) isolated in Korea. The comparisons of the nucleotide sequence of the cloned CP gene and its deduced amino acid sequences with other CP genes revealed that the CMV-ABI belongs to subgroup I (type I), CMV-ABI developed the typical mosaic symptom in infected plants. Tobacco plants (Samsun and NC82) were transformed by leaf-disc transformation via Agrobacterium, temefaciens LB4404 harboring pVCP, witch CMV-ABI CP gene was inserted into the pBI121, and a number of mature transgenic tobacco plants were developed. Southern and PCR analysis of genomic DNA from the transgenic plants showed that the CP gene was integrated into the genomes of the most of the transgenic plant. Result of the segregation patterns of resistance in T1 seedlings of the plants to kanamycin showed that the transgenic plants containing l,2 and 3 copies of CP gene were50%, 39% and 11% of the total transgenic plants, respectively.

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Evaluating Cultured Sea Mussels Mytilus edulis Extractions Methods and Extract Quality Characteristics (진주담치(Mytilus edulis) 추출물의 제조 및 품질특성)

  • Kim, Seon-Geun;Cho, Jun-Hyun;Hwang, Young-Sook;Lee, In-Seok;Oh, Kwang-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.6
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    • pp.650-655
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    • 2017
  • Extraction methods for cultured sea mussels Mytilus edulis and the quality characteristics of resulting extracts were investigated. The crude protein, carbohydrate and volatile basic nitrogen content of raw sea mussels was 15.2%, 1.9%, and 11.2 mg/100 g, respectively. Extracts were prepared using three different methods: hot-water extract (WE), scrap enzymatic hydrolysate extraction (SE), and complex extraction (CE). The respective extracts contained 5.5%, 8.6%, and 6.6% crude protein; 281.7, 366.0, and 343.0 mg/100 g amino nitrogen,: and 2.0%, 1.1% and 1.8% salinity. Their extraction yields were 689, 323, and 1,012 mL/kg. The CE method was superior to the traditional WE method in terms of extraction yield, amino-nitrogen content, and organoleptic qualities, but not odor. Active taste components were evaluated and the total free amino acid content of the WE and CE methods was 5,667.0 and 7,006.3 mg/100 g, respectively. The concentrations of major components (for WE and CE methods, respectively) were as follows: glutamic acid (1,244.0 and 955.4 mg/100 g), taurine (987.9 and 746.8 mg/100 g), glycine (721.2 and 847.0 mg/100 g), alanine (341.9 and 423.8 mg/100 g), arginine (265.5 and 376.5 mg/100 g), lysine (199.8 and 270.4 mg/100 g), and proline (253.9 and 220.3 mg/100 g). In conclusion, these results demonstrate that there is potential for using the CE method to expand the commercial utilization of sea mussels as a flavoring substance resource.

Study on the Identification and Contents of New Amino Acid in Edible Mushrooms (식용(食用)버섯중(中)의 새로운 아미노산(酸)의 구명(究明))

  • Ro, Ihl-Hyeob
    • Journal of Nutrition and Health
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    • v.12 no.1
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    • pp.31-41
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    • 1979
  • Free amino acid in ethanol extracts and total amino acids in hydrolysates of eleven species of edible mushrooms were analyzed and determinated the contents five kind of new amino acid by means of amino acid autoanalyzer and gas liquid chromatography. The result obtained from this study are as follows. 1) Five kind of new amino acid turned out to be ${\alpha}$-aminobutyric acid, allo-isoleucine, ethanolamine, $\gamma$-aminobutyric acid and ornithine. 2) By means of amino acid autoanalyzer, the monoethanolamine was identified on the chromatogram ahead of alanine, ${\alpha}$-aminobutyric acid between peak of threonine and glycine, allo-isoeleucine between peak of valine and leucine, isoleucine, ${\gamma}$-aminobutyric acid followed by proline between peak of leucine, isoleucine and methionine and ornithine between peak of phenylalanine and tyrosine 3) By means of Gas liquid chromatography, the ${\alpha}$-aminobutyric acid was identified on the chromatogram between peaks of alanine and valine, allo-isoleucine between peaks of methionine and isoleucine, monoethanolamine followed by ${\gamma}$-aminobutyric acid between peaks of phenylalanine and ammonia, ornithine between the peaks of ammonia and lysine. 4) Of five amino acids which were identified, ornithine was the highest of its content in the mushroom extracts, and allo-isoleucine, ethanolamine, and ${\gamma}$-aminobutyric acid came next in decreasing order. 5) Also which were identified, ornithine was the highest of its content in the hydrolysates, and ${\alpha}$-aminobutyric acid, ${\gamma}$-aminobutyric acid, allo-isoleucine came next in decreasing order, ethanol extracts and hydrolysates of Auriculariaauricula-Judae(Fr.) $Qu\acute{e}l$ species didn't contain any of five kind of new amino acid. Ornithine also was the highest in the hydrolysates of ll mushrooms.

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Taste-active Components of Powdered Smoke-dried Oysters and Its Application (훈건 굴을 이용한 분말조미소재의 정미성분 및 활용)

  • Kong Cheong-Sik;Kang Su-Tae;Ji Seung-Gil;Kang Jeong-Goo;Choi Dong-Jin;Kim Jeong-Gyun;Oh Kwang-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.39 no.3
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    • pp.278-282
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    • 2006
  • The powdered boil-dried oyster (C), the powdered smoke-dried oyster (OS-1) and the powdered smoke-dried oyster scrap (OS-2) were examined for their taste-active components, quality characteristics for potential utilization as a natural flavoring substance. The free amino acid level of OS-l and OS-2 were 395.6 mg/100 g and 551.5 mg/100 g, respectively, and that of betaine of OS-1 and OS-2 were 164.6 mg/100 g and 214.9 mg/100 g, respectively. The contents of inorganic ions were rich in Na, K, P, Ca and Mg in that order. Major free amino acids were taurine, glutamic acid, proline, alanine and glycine. The extract condition of the OS-1 and OS-2 the instant soup was the most appropriate at $98^{\circ}C$ for 1-5 min. The hot water extract of OS-1 and OS-2 with additives such as salt, sugar, pepper and onion powder had a good organoleptic qualities compared with the conventional flavoring substances in a local market. We conclude that powdered smoke-dried oyster and its scrap can be commercialized.

Composition of Free Amino Acids and Essential Oils in Root of Anthriscus sylvestylis (전조 뿌리의 유리 아미노산과 정유 성분 조성)

  • 김상국;권태용;민기군;이승필;최부술;이상철
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.41 no.5
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    • pp.521-525
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    • 1996
  • The study was carried out to find compositions of proximate components, free amino acid, and essential oils from root of Anthriscus sylvestylis. Proximate component contents were 7.69% for protein, 1.74% for fat, 2.44% for fiber, and 3.76% for ash. Extract content was 27.68% in fresh root. The compositions of free amino acids consisted 16 kinds. Phenylalanine content was the highest in composition of free amino acids. The essential oils of the root of Anthriscus sylvestylis was examined. $\alpha$-pinene, campreol, ,$\beta$-pinene, sabinene, myrcene, phellandrene, $\alpha$-terpinolene, d-limone, ${\gamma}$-terpinene, p-cymene, $\alpha$-terpinolene, carboxaldehyde, 3-cyc1ohexen-l-carboxaldehyde, 2-nonenal, isobornyl acetate, 4-terpineol, $\beta$-bisabolene, cis-piperitol, p-cymen-8-ol, BHT, methyl eugenol and 2-methoxy-4-vinyl-phenol were identified from the diethylether layers. Recovery yield of essential oils of Anthriscus sylvestylis of root was 0.58%. As a result, it was considered that the plant is worthy of cultivating as spice and medicinal crops.

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Changes in Solution Properties of Poly(trans-5-methylproline) During Mutarotation (Poly(trans-5-methylproline)의 변광회전에 따른 용액의 성질변화)

  • Han Man Jung
    • Journal of the Korean Chemical Society
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    • v.23 no.1
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    • pp.7-14
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    • 1979
  • Changes in CD, ORD and uv-spectra during the mutarotation of poly(trans-5-methyl-L-proline) (PTMP) were studied. The forward mutarotion of PTMP occurred in strong organic acids and trifluoroethanol, while the reverse mutarotation was observed by dilution of the trifluoroethanol solution with excess aliphatic alcohols. The changes in CD, ORD and uv-spectra during the forward and reverse mutarotation proceeded paralell to those found for the mutarotation of polyproline. The chemical shift of the ${\alpha}CH-$proton was shifted downfield about 0.3 ppm during the forward mutarotation. The reduced viscosity for the forward mutarotation increased from 0.15 to 0.26 (dl/g) during 5 days. The equilibrium between form I and form II was estabilished in an appropriate solvent mixture. All changes in solution properties mentioned above are similar to those found for polypoline. These results support that the two forms of PTMP are the same conformations as polyproline form I and form II, i. e., a right-handed helix with all cis amide bonds and a lefthanded helix with all trans amide bonds.

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ACE Inhibitory and Antihypertensive Effect of Chitosan Oligosaccharides in SHR (Chitosan 올리고당의 안지오텐신 전환효소 활성 억제 및 SHR에서의 고혈압 억제 특성)

  • Hong, Sang-Pill;Kim, Myung-Hee;Oh, Se-Wook;Han, Chan-Kyu;Kim, Yong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.30 no.6
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    • pp.1476-1479
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    • 1998
  • Effect of chitosan oligosaccharides on the ACE (angiotensin I converting enzyme) inhibition and antihypertension in SHR (Spontaneously hypertensive rat) was examined. The ACE inhibition activity was observed in all the chitosan oligosaccharides used in this study, and chitosan trimer exhibited the highest inhibitory activity $(IC_{50}=0.9{\mu}M)$ compared with other chitosan oligosaccharides $(IC_{50}\;:\;2.4{\sim}100\;{\mu}M)$. The results suggested that chitosan trimer was a good inhibitor of ACE in molecular level. When the single oral dose (2.14 mg/kg, similar to dose level of Captopril, known as strong ACE inhibitor) of chitosan trimer was given to 8 or 21 week aged SHR, the blood pressure reduction of both SHRs in 4hrs were $27{\pm}4.8\;mmHg\;and\;36{\pm}4.3\;mmHg$, respectively. Therefore, it was suggested that chitosan trimer could be applicable as natural ACE inhibitor related to antihypertension.

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