Partial purification and characterization of ribonuclease III like enzyme activity from cultured mouse embryo cells

Biochemistry ◽  
1978 ◽  
Vol 17 (23) ◽  
pp. 5052-5057 ◽  
Author(s):  
G. Shanmugam
Keyword(s):  
Enzyme Activity ◽  
Mouse Embryo ◽  
Partial Purification ◽  
Purification And Characterization ◽  
Ribonuclease Iii ◽  
Embryo Cells

scholarly journals Partial purification and characterization of an endo-α-N-acetylgalactosaminidase from the culture medium of Streptomyces sp. OH-11242

1992 ◽  
Vol 288 (2) ◽  
pp. 475-482 ◽  
Author(s):  
I Ishii-Karakasa ◽  
H Iwase ◽  
K Hotta ◽  
Y Tanaka ◽  
S Omura
Keyword(s):  
Enzyme Activity ◽  
Culture Medium ◽  
Gel Filtration ◽  
Partial Purification ◽  
Gastric Mucus ◽  
Purification And Characterization ◽  
Streptomyces Sp ◽  
Optimum Ph ◽  
New Type

For the purification of a new type of endo-alpha-N-acetylgalactosaminidase from the culture medium of Streptomyces sp. OH-11242 (endo-GalNAc-ase-S) [Iwase, Ishii, Ishihara, Tanaka, Omura & Hotta (1988) Biochem. Biophys. Res. Commun. 151, 422-428], a method for assaying enzyme activity was established. Using purified pig gastric mucus glycoprotein (PGM) as the substrate, oligosaccharides liberated from PGM were pyridylaminated, and the reducing terminal sugars of oligosaccharides larger than Gal beta 1-3GalNAc were analysed by h.p.1.c. The crude enzyme of endo-GalNAc-ase-S was prepared as an 80% (w/v) ammonium sulphate precipitate from the concentrated culture medium. The enzyme was partially purified by gel chromatofocusing and subsequent DEAE-Toyopearl chromatography. Endo-enzyme activity eluted around pI 4.8 on a gel chromatofocusing column and eluted with 0.19-0.25 M-NaCl on a DEAE-Toyopearl column. In the enzyme fraction obtained, no exo-glycosidases or proteases could be detected. The molecular mass of the enzyme was estimated as 105 kDa by gel filtration, and the optimum pH was 5.5. Endo-GalNAc-ase-S hydrolysed the O-glycosidic linkage between GalNAc and Ser (Thr) in 3H-labelled and unlabelled asialofetuin, liberating both the disaccharide (Gal beta 1-3GalNAc) and the tetrasaccharide [Gal beta 1-3 (Gal beta 1-4GlcNAc beta 1-6)GalNAc]. When endo-alpha-N-acetylgalactosaminidase from Alcaligenes sp. (endo-GalNac-ase-A) was incubated with 3H-labelled and unlabelled asialofetuin, only the disaccharide (Gal beta 1-3GalNAc) was liberated.

scholarly journals Optimization of Growth Conditions for Purification and Production of L-Asparaginase bySpirulina maxima

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Hanaa H. Abd El Baky ◽  
Gamal S. El Baroty
Keyword(s):  
Enzyme Activity ◽  
Gel Filtration ◽  
Growth Conditions ◽  
Partial Purification ◽  
Specific Enzyme ◽  
Purification And Characterization ◽  
Spirulina Maxima ◽  
Optimum Ph ◽  
Biomass Yields

L-asparaginase (L-AsnA) is widely distributed among microorganisms and has important applications in medicine and in food technology sectors. Therefore, the ability of the production, purification, and characterization of AsnA fromSpirulina maxima(SM) were tested. SM cultures grown in Zarrouk medium containing different N2(in NaNO3form) concentrations (1.25, 2.50, and 5.0 g/L) for 18 days contained a significant various quantity of dry biomass yields and AsnA enzyme levels. MS L-AsnA activity was found to be directly proportional to the N2concentration. The cultures of SM at large scales (300 L medium, 5 g/L N2) showed a high AsnA enzyme activity (898 IU), total protein (405 mg/g), specific enzyme activity (2.21 IU/mg protein), and enzyme yield (51.28 IU/L) compared with those in low N2cultures. The partial purification of crude MS AsnA enzyme achieved by 80% ammonium sulfate AS precipitated and CM-Sephadex C-200 gel filtration led to increases in the purification of enzyme with 5.28 and 10.91 times as great as that in SM crude enzymes. Optimum pH and temperature of purified AsnA for the hydrolyzate were 8.5 and 37 ± 0.2°C, respectively. To the best of our knowledge, this is the first report on L-asparaginase production inS. maxima.

Partial purification and characterization of phosphoenolpyruvate carboxylase from the cyanobacterium Anabaena variabilis

1986 ◽  
Vol 64 (5) ◽  
pp. 427-433 ◽  
Author(s):  
Nora W. Lem ◽  
Donna M. Penrose ◽  
Bernard R. Glick
Keyword(s):  
Enzyme Activity ◽  
Molecular Mass ◽  
Polyacrylamide Gel Electrophoresis ◽  
Anabaena Variabilis ◽  
Partial Purification ◽  
Purification And Characterization ◽  
Carboxylase Activity ◽  
Pep Carboxylase ◽  
Cyanobacterium Anabaena

The partial purification and characterization of phosphoenolpyruvate (PEP) carboxylase from the cyanobacterium Anabaena variabilis is reported. Spheroplasts made from photoautotrophically grown cells were lysed to produce a crude cell extract which was fractionated by (i) (NH4)2SO4 precipitation and (ii) gel filtration through Sephacryl S-300. The peak of enzyme activity that was eluted from the column corresponded to a molecular mass of approximately 365 000 daltons; the molecular mass of the sub-units was found to be approximately 100 500 daltons as assessed by polyacrylamide gel electrophoresis. The pH optimum for PEP carboxylase activity was pH 7.0. The response of this A. variabilis enzyme to effectors distinguished this enzyme from that found in C3 plants and in bacteria. Malate inhibited enzyme activity but was a much more effective inhibitor at pH 7.0 (50% inhibition at 1.6 mM malate) than at pH 8.0 (15% inhibition at 10 mM malate). Glycine stimulated PEP carboxylase activity at both pH 7.0 and 8.0, while glucose 6-phosphate had no significant effect. The Km for HCO3− was calculated to be 14.5 ± 9.2 μM. These results suggest that PEP carboxylase from A. variabilis is kinetically similar to maize PEP carboxylase and may participate in inorganic carbon uptake for photosynthesis.

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