• Title/Summary/Keyword: L-tyrosine

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Purification and Characterization of Protease from the Hepatopancreas of Shrimp, Penaeus orientalis

  • Oh Eun-Sil;Kim Doo-Sang;Choi Sung-Mi;Kim Jeong-Han;Pyeun Jae-Hyeung;Cho Deuk-Moon;Kim Hyeung-Rak
    • Fisheries and Aquatic Sciences
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    • v.2 no.2
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    • pp.218-225
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    • 1999
  • A protease without tryptic and chymotryptic activities was purified from the hepatopancreas of shrimp, Penaeus orientalis, using Q-Sepharose ionic exchange, benzamidine Sepharose-6B affinity, Mono-Q, and gel chromatography. Molecular weight (M.W.) of the protease was estimated to be 27kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS­PAGE). The amino acid composition of the protease was different from that of protease from P. japonicus or trypsin from P. orientalis. The protease was completely inhibited by benzamidine, $N\alpha-p-tosyl-L-lysine$ chloromethyl ketone (TLCK), and phenylmethylsulfonyl fluoride (PMSF) and was not affected by leupeptin, pepstatin, N-tosyl-L-phenylalanine chloromethyl ketone (TPCK), iodoacetate, and ethylenediamine tetra acetate (EDTA). The enzyme did not have any activity against Na-benzoyl-DL-arginine p-nitroanilide (BAPNA) or N-benzoyl-L-tyrosine ethyl ester (BTEE) which are specific substrates of trypsin and chymotrypsin, respectively. However, the protease showed hydrolytic activity for a carboxyl terminal of Tyr, Trp, Phe, Glu, and Cys.

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Recent Advances in Tyrosinase Research as An Industrial Enzyme (산업용 효소로써 티로시나아제 연구의 최근 동향)

  • Kim, Hyerin;Kim, Hyunmi;Choi, Yoo Seong
    • KSBB Journal
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    • v.29 no.1
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    • pp.1-8
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    • 2014
  • Tyrosinases catalyze the hydroxylation of monophenolic compounds and the conversion of o-diphenols to oquinones. The enzymes are mainly involved in the modification of tyrosine into L-3,4-dihydroxyphenyl-alanine (L-DOPA) and DOPA/DOPAquinone-drived intermolecular cross-linking, which play the key roles of pigmentation to the cells. It is ubiquitously distributed in microorganisms, plants, and animals all around the nature world. They are classified as copper- containing dioxygen activating enzymes; two copper ions are coordinated with six histidine residues in their active sites and they are distinguished as met-, deoxy-, and oxy-form depending on their oxidative states. Natural extraction and recombinant protein approaches have been tried to obtain practical amounts of the enzymes for industrial application. Tyrosinases have been widely applied to industrial and biomedical usages such as detoxification of waste water containing phenolic compounds, L-DOPA as a drug of Parkinson's disease, biomaterials preparation based on the cross-linking ability and biosensors for the detection of phenolic compounds. Therefore, this review reports the mechanism of tyrosinase, biochemical and structural features and potential applications in industrial field.

Isolation and Identification of Lactic Acid Bacteria with Probiotic Activities from Kimchi and Their Fermentation Properties in Milk (전통 김치로부터 Probiotic 유산균의 분리 및 우유 발효 특성)

  • Lim, Young-Soon;Kim, JiYoun;Kang, HyeonCheol
    • Journal of Dairy Science and Biotechnology
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    • v.37 no.2
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    • pp.115-128
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    • 2019
  • Lactic acid bacteria obtained from traditional Kimchi were selected on the basis of their caseinolytic activity and lactose usability and examined for availability as a starter in probiotic activity. Thirty-two strains were selected as lactic acid producing bacteria in BCP agar, and two strains (KC23 and KF26) with more than 90% resistance for both acid and bile salts were selected. The two strains were identified as L. plantarum (KC23) and L. paracasei (KF26) by API 50 CHL system and 16S rRNA sequence analysis. L. plantarum (KC23) was finally selected based on its biochemical characteristics for lactose and raffinose usability. Free tyrosine content increased rapidly in 10% skimmed milk medium, from $24.1{\mu}g/mL$ after 8 h to $43.9{\mu}g/mL$ after 16 h. Additionally, the caseinolytic clear zone of 12 mm of L. plantarum (KC23) was greater than the 9 mm zone of commercial L. acidophilus CSLA. The bacterium exhibited mesophilic growth and yielded $8.9{\times}10^8CFU/mL$ when incubated at $37^{\circ}C$ for 12 h at pH 4.25. Moreover, L. plantarum KC23 exhibited antibacterial activity as it formed a clear zone of 8-13 mm for the 5 pathogens. Adherent activity was 2.23 fold higher than that of LGG. The acidity of 10% skimmed milk fermented for 12 h was 0.74%.

Quality Characteristics of Soybean Curd Mixed with Freeze Dried Onion Powder (동결건조 양파분말을 첨가한 두부의 품질특성)

  • Kang, Nan-Suk;Kim, Jun-Han;Kim, Jong-Kuk
    • Food Science and Preservation
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    • v.14 no.1
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    • pp.47-53
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    • 2007
  • Soybean curd was mixed with onion powder to develop new foods, and changes in quality characteristics were investigated. The moisture content of onion soybean curd rose as the proportion of onion powder increased Whiteness (as measured by the L value) was high in soybean cud admixed with 0.1% (w/v) onion powder, Redness (the a value) was not significantly altered (the readings were 1.03-1.54) on addition of various onion powder concentrations. Yellowness (the b value) was similarly unaffected (readings 13.00-13.93) when various levels of onion powderwere added. Free sugar analysis showed that glucose was high in soybean curd (67.22 g/100 g) admixed with 0.1% (w/v) onion powder, The main organic acid was tartaric acid, and control organic acids included citric and oxalic acids at high levels. The major free amino acids were L-arginine, ${\gamma}-Amino-n-butyric$ acid, L-histidine, L-glutamic acid, L-serine, L-tyrosine and L-threonine, and amino acid content were high in soybean curd admixed with 0.2% (w/v) onion powder. Major phenolic compounds of onion soybean curd were quercitrin, protocatechuic acid and caffeic acid. The hardness of onion soybean curd was similar to that of the control when onion powder was added to 0.1% or 0.2% (w/v), and decreasedmore onion powder was added. Organoleptic qualities dropped as onion powder levels increased. In summary, onion powder addition to soybean curd is optimal at the 0.2% (w/v) level..

Antioxidant activity and physicochemical composition of fermented Vigna angularis using Bacillus subtilis KCCM 11965P (Bacillus subtilis KCCM 11965P를 이용한 붉은팥 발효물의 항산화 활성 및 이화학성분 분석)

  • Jeong, Kyung Ok;Oh, Keun Su;Moon, Kwang Hyun;Kim, Dae Geun;Im, So Yeon;Lee, Eun Ji;Kim, Na Ri;Kim, Wook;Kim, Hae Jin;Lee, Jeong Ho
    • Food Science and Preservation
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    • v.24 no.7
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    • pp.975-982
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    • 2017
  • Health of human intestine has deteriorated due to excessive stress and western diet. In this study, Vigna angularis was fermented by Bacillus subtilis KCCM11965P in order to develop prebiotic resource for improving bowel movement. The contents of ash and crude protein were $3.35{\pm}0.04%$ and $21.1{\pm}0.19%$ respectively. Vigna angularis extract (1, 3, 5%) were incubated with 3% (v/v) Bacillus subtilis KCCM 11965P for 0, 24, 48, and 72 h. Total bacterial numbers showed that the combination of 3% powdered seeds and 72 h incubation time was optimum condition for this experiment. Total polyphenol content increased from $0.18{\pm}0.010mg/mL$ in pre-incubation to $0.23{\pm}0.007mg/mL$ in post-incubation with the condition mentioned above. DPPH radical scavenging activity also increased from $36.1{\pm}6.0%$ to $63.6{\pm}5.2%$. Analysis of protease activity showed $2.69{\pm}0.003unit/mL$ in combination of 5% powdered seeds and 72 h incubation time. Amylase activity increased from $1.0{\pm}0.1unit/mL$ in pre-incubation to $26.0{\pm}0.2unit/mL$ in post-incubation. The analysis of free amino acids after incubation with Bacillus subtilis KCCM 11965P showed that leucine increased from 5.22 mg/L to 67.59 mg/L and tyrosine, one of non-essential amino acid also increased 10.08 mg/L to 259.35 mg/L by incubation with 5% powdered seeds. Most of organic acid were reduced by incubation for 72 h. These results suggest that Vigna angularis could be utilized most as a prebiotic resources.

Purification and Characterization of Polyphenol Oxidase from Burdock (Arctium lappa L.) (우엉(Arctium lappa L.) 뿌리 Polyphenol Oxidase의 부분정제 및 특성)

  • Lim, Jeong-Ho;Jeong, Moon-Cheol;Moon, Kwang-Deog
    • Food Science and Preservation
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    • v.12 no.5
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    • pp.489-495
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    • 2005
  • Polyphenol oxidase (PPO) from Burdock (Arctium lappa L.) was purified and characterized. Purification of polyphenol oxidase was achieved by ammonium sulfate precipitation, Phenyl-sepharose CL-4B hydrophobic chromatography and Sephadex G-100 gel filtration chromatography. The molecular mass of the purified PPO was estimated to be 30 kDa by SDS polyacrylamide gel electrophoresis. In a substrate specificity, maximum activity was achieved with chlorogenic acid, followed by catechol and catechin. Whereas, there was low activity with hydroquinic acid, resorcinol or tyrosine. The optimum pH and temperature for enzyme activity were 7.0 and 35$\circC$ with catechol, respectively. The enzyme was most stable at pH 7.0 while unstable at acidic and alkaline pH. The enzyme was stable when heated to 40$\circC$. But heating at 50$\circC$ for more than 30 min caused 50% loss of activity. Ascorbic acid, L-cystein and $Cu^{2+}$ inhibited the activity of pholyphenol oxidase.

Changes of Chemical Components during Fermentation of Pear Wine (배술 발효 과정 중 화학 성분의 변화)

  • Lee, Ka-Soon;Park, Hae-Min;Hong, Jong-Sook;Lee, Gyu-Hee;Oh, Man-Jin
    • Food Science and Preservation
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    • v.16 no.6
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    • pp.991-998
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    • 2009
  • We used pears to manufacture wine, and analyzed changes in pH, acidity and ethanol and sugar content during fermentation. Pear wine with added ginger (to improve quality) did not differ from ginger-free wine in pH or acidity level. The ethanol content of pear wine was the highest (13.0%, v/v) inpear wine with 0.1% (w/v) added ginger compared to pear wine with no ginger, and sensory tests examining taste and color yielded the highest scores for pear wine with 0.2% (w/v) ginger. To assess storage stability, pear wine was treated for 30 minutes at $55^{\circ}C$, $60^{\circ}C$, $65^{\circ}C$, or $70^{\circ}C$. Unheated pear wine showed rapid changes in pH and acidity level after 30 days of storage, whereas pear wine treated for 30 minutes at $60^{\circ}C$ did not show such changes. Total organic acid levels in pear wine increased by 0.71% and 0.89% (v/v), respectively. The free sugar level in pear wine decreased from 12.05% to 3.13% (w/v). Turning to phenolic compounds, caffeic acid, catechin, and epicatechin contents in pears were 1.64, 1.40, and 0.23 mg/100mL, respectively, with diverse compositions. Caffeic acid levels in pear wine decreased sharply to 0.12 mg/100 mL upon fermentation, whereas free catechin inpear wine increased to 1.16 mg/100 mL compared with 0.28 mg/100 mL in pears. Free arbutin increased from 8.34 mg/100 mL in pears to 10.39 mg/100 mL in pear wine. The free amino acid content of pear wine was 118.5 g/100 mL, but the levels of serine, alanine, glutamic acid, and aspartic acid decreased sharply upon fermentation, with corresponding increases in tyrosine, GABA, lysine, and arginine.

Biosynthetic pathway of shikimate and aromatic amino acid and its metabolic engineering in plants (식물에서 shikimate 및 방향족 아미노산 생합성 경로와 이의 대사공학적 응용)

  • Lim, Sun-Hyung;Park, Sang Kyu;Ha, Sun-Hwa;Choi, Min Ji;Kim, Da-Hye;Lee, Jong-Yeol;Kim, Young-Mi
    • Journal of Plant Biotechnology
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    • v.42 no.3
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    • pp.135-153
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    • 2015
  • The aromatic amino acids, which are composed of $\small{L}$-phenylalanine, $\small{L}$-tyrosine and $\small{L}$-tryptophan, are general components of protein synthesis as well as precursors for a wide range of secondary metabolites. These aromatic amino acids-derived compounds play important roles as ingredients of diverse phenolics including pigments and cell walls, and hormones like auxin and salicylic acid in plants. Moreover, they also serve as the natural products of alkaloids and glucosinolates, which have a high potential to promote human health and nutrition. The biosynthetic pathways of aromatic amino acids share a chorismate, the common intermediate, which is originated from shikimate pathway. Then, tryptophan is synthesized via anthranilate and the other phenylalanine and tyrosine are synthesized via prephenate, as intermediates. This review reports recent studies about all the enzymatic steps involved in aromatic amino acid biosynthetic pathways and their gene regulation on transcriptional/post-transcriptional levels. Furthermore, results of metabolic engineering are introduced as efforts to improve the production of the aromatic amino acids-derived secondary metabolites in plants.

Synthesis, Crystal Structure and Theoretical Calculation of a Novel Nickel(II) Complex with Dibromotyrosine and 1,10-Phenanthroline

  • Huang, Guimei;Zhang, Xia;Fan, Yuhua;Bi, Caifeng;Yan, Xingchen;Zhang, Zhongyu;Zhang, Nan
    • Bulletin of the Korean Chemical Society
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    • v.34 no.10
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    • pp.2889-2894
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    • 2013
  • A new complex [$Ni(phen)(C_9H_8Br_2NO_3)_2{\cdot}2CH_3OH{\cdot}2H_2O$] [phen: 1,10-phenanthroline $C_9H_8Br_2NO_3$: 3,5-dibromo-L-tyrosine] was synthesized and characterized by IR, elemental analysis and single crystal X-ray diffraction. X-ray crystallography shows that Ni(II) ion is six-coordinated. The Ni(II) ion coordinates with four nitrogen atoms and two oxygen atoms from three ligands, forming a mononuclear Ni(II) complex. The crystal crystallizes in the Orthorhombic system, space group $P2_12_12$ with a = 12.9546 ${\AA}$, b = 14.9822 ${\AA}$, c = 9.9705 ${\AA}$, V = 1935.2 ${\AA}$, Z = 1, F(000) = 1008, S = 0.969, ${\rho}_{calcd}=1.742g{\cdot}cm^{-3}$, ${\mu}=4.688mm^{-1}$, $R_1$ = 0.0529 and $wR_2$ = 0.0738 for 3424 observed reflections (I > $2{\sigma}(I)$). Theoretical study of the title complex was carried out by density functional theory (DFT) method and the B3LYP method employing the $6-3l+G^*$ basis set. The energy gap between HOMO and LUMO indicates that this complex is prone to interact with DNA. CCDC: 908041.

Nutritional and Agronomic Characteristics of Super-Giant Embryo Mutant in Rice (벼 초거대배 돌연변이계통의 영양 및 작물학적 특성)

  • Koh, Hee-Jong;Park, Sun-Zik;Won, Yong-Jae;Heu, Mun-Hue
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.38 no.6
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    • pp.537-544
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    • 1993
  • Super-giant-embryo mutant line (Hwacheong-ge$^s$) with high nutritional quality was developed from Hwacheongbyeo, a Korean Japonica cultivar. Grain weight of brown rice of Hwacheong-ge$^s$ line was lighter than of Hwacheongbyeo. However, embryo dry weight of the line was 3.22 times heavier than that of Hwacheongbyeo on the single grain base. Hwacheong-ge$^s$ line showed higher protein(8.99%) than the original variety(7.39%), without changes in storage protein. In amino acid composition, lysine content was greatly increased in the mutant, while the contents of methionine, serine and tyrosine were slightly decreased as compared with the original one. Lipid content of Hwacheong-ge$^s$ line was 1.5 times higher compared with that of Hwacheongbyeo. The contents of Vitamin Bl, B2, and E(${\alpha}$-tocopherol) were also greatly increased in Hwacheong-ge$^s$line. The grain yields of the mutant lines ranged from 69.2% to 78.8% compared with that of Hwacheongbyeo, 556kg /10a. Most of the mutant lines of M4 generation were homogeneous for growth characters, and some of them seemed to be readily applicable for farmer's field.

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