Abstract
An arginine specific protease, Sp-protease, was purified by column chromatography from freeze-dried Spirulina platensis using a five-step process. Purified Sp-protease has a molecular weight of 80 kDa. It hydrolyzed the synthetic substrates containing arginine residue in the P1 position but did not hydrolyze synthetic substrates containing other amino acid residues, including lysine residue in the P1 position. Among the synthetic substrates tested, a substrate of plasminogen activator (Pyr-Gly-Arg-MCA) was hydrolyzed most effectively with the enzyme (K m = 5.5 × 10−6 M), and fibrin gel was solubilized via activation of intrinsic plasminogen to plasmin with the enzyme. Activity was inhibited completely with camostat mesilate (K i = 1.1 × 10−8 M) and leupeptin (K i = 3.9 × 10−8 M) but was not inhibited with Nα-tosyl-L-lysine chloromethyl ketone (TLCK). The optimum pH of the enzyme has a range of pH 9.0 to pH 11.0. The optimum temperature was 50°C; the enzyme was stable at 0–50°C.
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Bradford MM (1976) A rapid sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254
Clement C, Lonchamp D, Rebeller M, Van Landeghem. H (1980) The development of Spirulina algae cultivation. Chem Eng Sci 35: 119–126
Dunlop JSR, Muir. MD, Milne VA, Groves. DI (1978) A new Dunlop, microfossil assemblage from the Archaean of Western Australia. Nature 274: 676–678
Iwamoto E, Kawanobe H, Shirawa Y, Ikawa T (2001) Purification and characterization of Mannito-I-phosphatase in the Red alga caloglossa continua (Ceramiales, Rhodophyta). Mar Bioteclmol 3: 493–500
Kadowaki T, Yoneda M, Okamoto K, Maeda K, Yamamoto K (1994) Purification and characterization of a novel arginirie- specific cystein proteinase (argingipain) involved in the pathogenesis of periodontal disease from the culture supernatant of Porphyromonas gingivalis, J Biol Chem 269: 21371–21378
Laemmli UK, (1970) Cleavage of structural proteins during the assembly of the head of basteriophage T4. Nature 227: 680–685
Lockau VL, Massalsky B, Dirmeier A (1988) Purification and partial characterization of a calcium-stimulated protease from the cyanobacterium, Anabaena variabilis. Eur J Biochem 172: 433–438
Matsushima A, Shioya K, Kobayashi M, Kodera Y, Inada Y (1993) Activation of fibrinolysis with the protease from Dermatophagoides farinae. Thromb Haemost 70: 545
Nanni B, Balestreri E, Dainese E, Cozzani I, Felicioli R. (2001) Characterisation of a specific phycocyanin-hydrolysing protease purified from Spinrulina platensis. Microbiol Res 156: 259–266
Noguchi Y, Ishii A, Matsushima A, Haishi D, Yasumuro K, Moriguchi T, Wada T, Kodera Y, Hiroto M, Nishimura H, Sekine M, Inada Y (1999) Isolation of biopterin-α-glucoside from Spirulina (Arthrospira) platensis and its physiologic function. Mar Biotechnol 1: 207–210
Richter R, Hejazi M, Kraft R, Ziegler K, Lockau W (1999) Cyanophycinase, a peptidase degrading the cyanobacterial reserve material multi-L-arginyl-poly-L-aspartic acid (cyanophycin). Eur J Biochem 263: 163–169
Saito T, Ishikura H, Hada Y, Fukui K, Kodera Y, Matsushima A, Inada Y (2003) Photostabilization of phycocyanin and anthocyanin in the presence of biopterin-α–glucoside from Spirulina platensis under ultraviolet ray. Dyes Pigments 56: 203–207
Schenkein L, Levy M, Franklin EG, Frangione B (1977) Proteolytic enzymes from the mouse submaxillary gland. Specificity restricted to arginine residues. Arch Biochem Biophys 182: 64–70
Yamano N, Higashida N, Endo C, Sakata N, Fujishima S, Maruyama A, Higashihara T (2000) Purification and characterization of N-Acetylglucosamine-6-phosphate Deacetylase from a psychrotrophic marine bacterium, Alteromonas species. Mar Biotechnol 2: 57–64
Zhao J, Li L, Wu C, He RQ (2003) Hydrolysis of fibrinogen and plasminogen by immobilized earthworm fibrinolytic enzyme II from Eisenia fetida. Int J Biol Macromol 32: 165–171
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Yada, E., Nagata, H., Noguchi, Y. et al. An Arginine Specific Protease from Spirulina platensis. Mar Biotechnol 7, 474–480 (2005). https://doi.org/10.1007/s10126-004-4115-9
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DOI: https://doi.org/10.1007/s10126-004-4115-9