• Title/Summary/Keyword: Protein purification

Search Result 1,026, Processing Time 0.037 seconds

Purification and Identification of a Natural Antioxidant Protein from Fertilized Eggs

  • Yang, Shaohua;Wang, Lulu;Wang, Ying;Ou, Xiaoqian;Shi, Zhaoyuan;Lu, Chongchong;Wang, Wei;Liu, Guoqing
    • Food Science of Animal Resources
    • /
    • v.37 no.5
    • /
    • pp.764-772
    • /
    • 2017

  • Fertilized hen eggs are rich in a variety of bioactive ingredients. In this study, we aimed to obtain an antioxidant protein from fertilized eggs and the radical scavenging abilities on 1, 1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl radical (${\bullet}OH$), superoxide anion ($O^{2-}{\bullet}$) were used to evaluate the antioxidant activity of the purified protein. During 20 d of incubation, the radical scavenging ability of protein extracted from fertilized eggs exhibited significantly differences and the protein on day 16 showed higher antioxidant capacity. Based on this, the antioxidant protein of the samples on day 16 were isolated for the follow-up study. With a molecular weight 43.22 kDa, the antioxidant protein was purified by Diethylaminoethyl cellulose -52 (DEAE-52) column and Sephadex G-100. The LC-MS analysis showed that the purified protein molecular weight was 43.22 kDa, named D2-S. The sequence of amino acids was highly similar to ovalbumin and the coverage reached to 84%. The purified protein showed a radical scavenging rate of $52.34{\pm}3.27%$ on DPPH and $63.49{\pm}0.25%$ on ${\bullet}OH$, respectively. Furthermore, the C-terminal amino acid sequence was NAVLFFGRCVSP, which was consistent with the sequence of ovabumin. These results here indicated that purified protein may be a potential resource as a natural antioxidant.

  • https://doi.org/10.5851/kosfa.2017.37.5.764 인용 PDF KSCI

Purification and characterization of Protease from Kyenegum (계내금(鷄內金) 단백질 분해 효소의 정제와 특성)

  • Kim, Do-Wan;Jo, Hye-Sim;Jeong, Yong-Jin;Kim, Kwang-Soo
    • The Korea Journal of Herbology
    • /
    • v.22 no.4
    • /
    • pp.21-28
    • /
    • 2007

  • Objectives : Kyenegum has been popularly used long as the digestive. The purpose of this study was to investigate the purification and characteristics of protease obtained from Kyenegum crude enzyme. Methods : Kyenegum protease was purified by precipitation with ammonium sulfate followed by SP-Spharose ion exchange chromatography. The molecular weight of Kyenegum protease was estimated by SDS-PAGE electrophoresis. Results : Kyenegum protease was 3,087 units/mg protein specific activity, 14.5 purification fold and 9.8 % recovery. The molecular weight of protease was estimated to be 18 kDa. The isoelectric point was pI 8.97 and values of Km and Vmax of its were 48 mg/mL and 2 units/min, respectively. Conclusion : The result suggests that the protease obtained from Kyenegum has excellent stability of temperature, acid and collagen substrate specificity.

  • PDF

Purification of Cold-adapted Protease from Janthinobacterium sp. (Janthinobacterium sp. 유래 저온 활성 프로테아제 정제)

  • Kim, Hyun-do;Choi, Jong-il
    • Microbiology and Biotechnology Letters
    • /
    • v.46 no.2
    • /
    • pp.111-113
    • /
    • 2018

  • In this study, purification of cold-adapted protease from Janthinobacterium sp. was investigated. First, using gradient precipitation, protease was confirmed to be deposited in the 30-80% range of ammonium sulfate. Next, DEAE-Sepharose column was used for the binding of the protease under various conditions. The optimal binding condition was found to be pH 8.5 and flow rate of 30 ml/h. Under the optimal condition, the protease was purified with 29% recovery yield. This result can be useful for the purification of other cold-adapted protein.

  • https://doi.org/10.4014/mbl.1802.02016 인용 PDF KSCI

Purification and Properties of Serratia marcescens Purine Nucleoside Phosphorylase. (Serratia marcescens Purine Nucleoside Phosphorylase의 정제 및 특성)

  • 방성권;신종란;최병범
    • Microbiology and Biotechnology Letters
    • /
    • v.28 no.5
    • /
    • pp.251-257
    • /
    • 2000

  • Serratia marcescens purine nucleoside phosphorylase (PNP) was purfied to homogeneity by streptomycin sulfate treatment, Sephacry HR S-200 gel filtration chromatography and AMP-agarose affinity chromatography. The specific activity of the enzyme was increased 49-fold during purification with an overall yield of 7.0%. The molecular weight was 168kD as estimated by Sephadex G-150 gel filtration chromatography. The S. marcescens enzyme was composed of six identical subunits with subunit molecular weight of 28kD, as estimated by SDS-PAGE. The Km values of S. marcescens enzyme for inosine and deoxyinsoine were 0.38 and 1.20 mM, respectively. The ph optimum was near 8.0, and the enzyme was relatively heat-stable protein. The enzyme was inactivated com-pletely by 0.5 mM of $Cu^{ 2+}$.

  • PDF