Purification and some properties of proteinase fromPseudomonas fluorescensNo. 33

1993 ◽  
Vol 60 (2) ◽  
pp. 229-237 ◽  
Author(s):  
Haruto Kumura ◽  
Katsuhiko Mikawa ◽  
Zenichi Saito
Keyword(s):  
Molecular Mass ◽  
Phosphate Buffer ◽  
Sodium Phosphate ◽  
Extracellular Proteinase ◽  
Original Activity ◽  
Sodium Phosphate Buffer ◽  
Sds Page ◽  
Optimum Ph ◽  
Hydrolysis Of ◽  
Optimum Ph And Temperature

SummaryThe extracellular proteinase fromPseudomonas fluorescensNo. 33 was purified to electrophoretic homogeneity by a procedure including precipitation with HC1 and (NH4)2SO4, and column chromatography. The enzyme was purified 170-fold giving a yield of 7 % of the original activity. The molecular mass of the purified enzyme was 48000 by SDS-PAGE. The optimum pH and temperature for the hydrolysis of casein were 8·0–9·8 and 30–35 °C respectively. The enzyme was more thermostable in synthetic milk salts solution than in 0·1 M-sodium phosphate buffer, but was heat-labile at 50 °C in both buffer Systems. The activity was inhibited byo−phenanthroline, Hg2+, Cu2+, Fe2+and, to a lesser extent, Ni2+. Caseins were susceptible to the proteinase, but degradation patterns were dependent on the form of the casein.

Effects of sodium phosphate buffer on horseradish peroxidase thermal stability

2008 ◽  
Vol 93 (2) ◽  
pp. 569-574 ◽  
Author(s):  
L. Haifeng ◽  
L. Yuwen ◽  
C. Xiaomin ◽  
W. Zhiyong ◽  
W. Cunxin
Keyword(s):  
Thermal Stability ◽  
Horseradish Peroxidase ◽  
Phosphate Buffer ◽  
Sodium Phosphate ◽  
Sodium Phosphate Buffer

scholarly journals Characterization of Biosynthetic Enzymes for Ectoine as a Compatible Solute in a Moderately Halophilic Eubacterium, Halomonas elongata

1999 ◽  
Vol 181 (1) ◽  
pp. 91-99 ◽  
Author(s):  
Hisayo Ono ◽  
Kazuhisa Sawada ◽  
Nonpanga Khunajakr ◽  
Tao Tao ◽  
Mihoko Yamamoto ◽  
...  
Keyword(s):  
Molecular Mass ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Apparent Molecular Mass ◽  
Optimum Temperature ◽  
Sds Page ◽  
Halomonas Elongata ◽  
Optimum Ph

ABSTRACT 1,4,5,6-Tetrahydro-2-methyl-4-pyrimidinecarboxylic acid (ectoine) is an excellent osmoprotectant. The biosynthetic pathway of ectoine from aspartic β-semialdehyde (ASA), in Halomonas elongata, was elucidated by purification and characterization of each enzyme involved. 2,4-Diaminobutyrate (DABA) aminotransferase catalyzed reversively the first step of the pathway, conversion of ASA to DABA by transamination with l-glutamate. This enzyme required pyridoxal 5′-phosphate and potassium ions for its activity and stability. The gel filtration estimated an apparent molecular mass of 260 kDa, whereas molecular mass measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was 44 kDa. This enzyme exhibited an optimum pH of 8.6 and an optimum temperature of 25°C and had Km s of 9.1 mM forl-glutamate and 4.5 mM for dl-ASA. DABA acetyltransferase catalyzed acetylation of DABA to γ-N-acetyl-α,γ-diaminobutyric acid (ADABA) with acetyl coenzyme A and exhibited an optimum pH of 8.2 and an optimum temperature of 20°C in the presence of 0.4 M NaCl. The molecular mass was 45 kDa by gel filtration. Ectoine synthase catalyzed circularization of ADABA to ectoine and exhibited an optimum pH of 8.5 to 9.0 and an optimum temperature of 15°C in the presence of 0.5 M NaCl. This enzyme had an apparent molecular mass of 19 kDa by SDS-PAGE and a Km of 8.4 mM in the presence of 0.77 M NaCl. DABA acetyltransferase and ectoine synthase were stabilized in the presence of NaCl (>2 M) and DABA (100 mM) at temperatures below 30°C.

Recovery of milk fat globule membrane (MFGM) from buttermilk: effect of Ca-binding salts

2019 ◽  
Vol 86 (3) ◽  
pp. 374-376 ◽  
Author(s):  
Vitaly L. Spitsberg ◽  
Liza Ivanov ◽  
Vladimir Shritz
Keyword(s):  
Phosphate Buffer ◽  
Milk Fat ◽  
Sodium Phosphate ◽  
Milk Fat Globule Membrane ◽  
Peripheral Protein ◽  
Sodium Phosphate Buffer ◽  
Milk Fat Globule ◽  
Fat Globule ◽  
First Time ◽  
Phosphate Groups

AbstractIn this Research Communication we present a study of the effect of Ca-binding salts on the recovery of milk fat globule membrane (MFGM) from buttermilk. Sodium phosphate buffer was used for the purpose of MFGM recovery from buttermilk for the first time and we showed that 0.1 M buffer at pH 7.2 was the most effective for the recovery of MFGM. The fact of high efficacy of sodium phosphate buffer in recovery of MFGM from buttermilk allowed us to suggest that MFGM in buttermilk is present in association with casein through Ca- bridges formed between phospholipids of MFGM and phosphate groups of casein, primarily with k-casein as the peripheral protein of casein micelles.

scholarly journals Purification and characterization of beta-1, 3-glucanase from the secretion of Simira glaziovii colleters (Rubiaceae)

2006 ◽  
Vol 49 (6) ◽  
pp. 881-888 ◽  
Author(s):  
Felipe Almeida Vieira ◽  
Maura da Cunha ◽  
Denise Espellet Klein ◽  
André de Oliveira Carvalho ◽  
Valdirene Moreira Gomes
Keyword(s):  
Molecular Mass ◽  
Purification Process ◽  
Purification And Characterization ◽  
Chromatographic Methods ◽  
Sds Page ◽  
Optimum Ph

In this study, beta-1,3-glucanase was isolated from Simira glaziovii secretion. The purification process was achieved by a combination of chromatographic methods and was analyzed by SDS-PAGE. The purified enzyme presented an estimated molecular mass of 35 kDa. The optimum pH of enzyme was 5.2

scholarly journals Large-scale purification of hepatitis B surface antigen

1976 ◽  
Vol 3 (6) ◽  
pp. 626-631
Author(s):  
J Vnek ◽  
A M Prince
Keyword(s):  
Polyethylene Glycol ◽  
Hepatitis B ◽  
Surface Antigen ◽  
Phosphate Buffer ◽  
Sodium Phosphate ◽  
Large Scale ◽  
Hepatitis B Surface Antigen ◽  
Sodium Phosphate Buffer ◽  
Subsequent Elution ◽  
Polyethylene Glycol Precipitation

Hepatitis B surface antigen was concentrated and purified from plasma by two simple steps of purification. In the first step the antigen was purified 24-fold by polyethylene glycol precipitation. An additional 10-fold purification was achieved by batchwise adsorption to hydroxylapatite and subsequent elution with 0.02 M sodium phosphate buffer.

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