scholarly journals Solubilization and other studies on adenylate cyclase of baker's yeast

1976 ◽  
Vol 159 (2) ◽  
pp. 363-370 ◽  
Author(s):  
K Varimo ◽  
J Londesborough
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cyclic Amp ◽  
Adenylate Cyclase ◽  
Gel Filtration ◽  
Baker’S Yeast ◽  
Baker's Yeast ◽  
Centrifugal Forces ◽  
Wide Range ◽  
Solubilized Enzyme ◽  
Fold Activation

1. Adenylate cyclase of Saccharomyces cerevisiae was sedimented from mechanically disintegrated preparations of yeast over an unusually wide range of centrifugal forces. 2. The enzyme was readily solubilized by Ficoll and by Lubrol PX. Lubrol caused a 2-fold activation. 3. Both particle-bound and Lubrol-solubilized enzyme had an apparent Km for ATP of 1.6 mM in the presence of 0.4 mM-cyclic AMP and 5 mM-MnCl2 at pH 6.2 and 30°C. 4. The Lubrol-solubilized enzyme behaved on gel filtration as a monodisperse protein with an apparent mol.wt. of about 450000.

scholarly journals Characterization of two trehalases in baker's yeast

1984 ◽  
Vol 219 (2) ◽  
pp. 511-518 ◽  
Author(s):  
J Londesborough ◽  
K Varimo
Keyword(s):  
Acetic Acid ◽  
Cyclic Amp ◽  
Concanavalin A ◽  
Soluble Fraction ◽  
Gel Filtration ◽  
Baker’S Yeast ◽  
Acidic Protein ◽  
Baker's Yeast ◽  
Ph Optimum

Trehalase activities at pH 5 (not inhibited by EDTA) and pH 7 (inhibited by EDTA) were present in the soluble fraction of disintegrated commercial baker's yeast. The pH 5 activity binds strongly to concanavalin A, is only partially salted out by saturated (NH4)2SO4, has an apparent Mr of 215000 (by gel filtration) and is an acidic protein. It has a Km of 1.4 mM, a broad pH optimum (at 40 mM-trehalose) between pH 4 and 5, and is activated by about 30% by 20-300 mM neutral salts such as KCl, NaNO3 and MnCl2. The enzyme is strongly inhibited by acetic acid/acetate buffers, with a Ki of about 15 mM-acetic acid. The pH 7 activity does not bind to concanavalin A, is salted out at 20-32% (w/v) (NH4)2SO4 and has an Mr of 170000 (by gel filtration). It is absolutely dependent on Ca2+ or Mn2+ ions (Mg2+ is ineffective) and strongly inhibited by neutral salts in the 20-100 mM range. It can be activated by treatment with MgATP in the presence of cyclic AMP. Activation decreases, but does not abolish, the Ca2+ requirement, and does not change the Km for trehalose (5.7 mM) or shift the sharp pH optimum at pH 6.7 (at 40 mM-trehalose).

scholarly journals Characterization of an adenosine 3′:5′-cyclic monophosphate phosphodiesterase from baker's yeast. Its binding to subcellular particles, catalytic properties and gel-filtration behaviour

1977 ◽  
Vol 163 (3) ◽  
pp. 467-476 ◽  
Author(s):  
J Londesborough
Keyword(s):  
Ionic Strength ◽  
Cyclic Amp ◽  
Microsomal Fraction ◽  
Gel Filtration ◽  
Potassium Phosphate ◽  
Apparent Molecular Weight ◽  
Baker’S Yeast ◽  
Baker's Yeast ◽  
Ph Optimum

1. The 3′:5′-cyclic AMP phosphodiesterase in the microsomal fraction of baker's yeast is highly specific for cyclic AMP, and not inhibited by cyclic GMP, cyclic IMP or cyclic UMP. Catalytic activity is abolished by 30 micrometer-EDTA. At 30 degrees C and pH8.1, the Km is 0.17 micrometer, and theophylline is a simple competitive inhibitor with Ki 0.7 micrometer. The pH optimum is about 7.8 at 0.25 micrometer-cyclic AMP, so that over the physiological range of pH in yeast the activity changes in the opposite direction to that of adenylate cyclase [PH optimum about 6.2; Londesborough & Nurminen (1972) Acta Chem. Scand. 26, 3396-3398].2. At pH 7.2, dissociation of the enzyme from dilute microsomal suspensions increased with ionic strength and was almost complete at 0.3 M-KCl. MgCl2 caused more dissociation than did KCl or NaCl at the same ionic strength, but at low KCl concentrations binding required small amounts of free bivalent metal ions. In 0.1 M-KCl the binding decreased between pH 4.7 and 9.3. At pH 7.2 the binding was independent of temperature between 5 and 20 degrees C. These observations suggest that the binding is electrostatic rather than hydrophobic. 3. The proportion of bound activity increased with the concentration of the microsomal fraction, and at 22 mg of protein/ml and pH 7.2 was 70% at I0.18, and 35% at I0.26. Presumably a substantial amount of the enzyme is particle-bound in vivo. 4. At 5 degrees C in 10 mM-potassium phosphate, pH 7.2, the apparent molecular weight of KCl-solubilized enzyme decreased with enzyme concentration from about 200 000 to 40 000. In the presence of 0.5M-KCl, a constant mol.wt. of about 55 000 was observed over a 20-fold range of enzyme concentrations.

The use of β-glucan extracted from baker’s yeast (Saccharomyces cerevisiae) to increase non-specific immune system and resistence of tilapia (Oreochromis niloticus) to Aeromonas hydrophila

2013 ◽  
pp. 92
Author(s):  
Ida N Jamal ◽  
Reiny A Tumbol ◽  
Remy E.P Mangindaan
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Immune System ◽  
Aeromonas Hydrophila ◽  
Phagocytic Activity ◽  
Baker’S Yeast ◽  
Baker's Yeast ◽  
Yeast Saccharomyces Cerevisiae ◽  
Randomized Design ◽  
The Difference ◽  
Motile Aeromonas Septicaemia

Motile Aeromonas Septicaemia disease (MAS) attacking tilapia has increased in recent years as a consequence of intensive aquaculture activities, which led to losses in aquaculture industry. The agent causing MAS disease is Aeromonas hydrophila. The disease can be controlled with the β-glucan. As immunostimulants, β-glucans can also increase resistance in farmed tilapia. Studies on the use of β-glucan extracted from baker's yeast Saccharomyces cerevisiae was intended to evaluate the non-specific immune system of tilapia that were challenged with Aeromonas hydrophila. The method used was an experimental method with a completely randomized design consisting of four treatments with three replicats. The dose of β-glucan used as treatments were 0 mg.kg-1 fish (Control), 5 mg.kg-1 fish (B), 10 mg.kg-1 fish (C) and 20 mg.kg-1 fish (D), each treatment as injected three times at intervals of 3 days, the injection volume of 0.5 ml/fish for nine days and resistance surveillance for seven days. The results showed that the difference in the amount of β-glucan and the frequency of the injected real influence on total leukocytes, phagocytic activity and resistance. Total leukocytes, phagocytic activity and resistance to treatment was best achieved by the administration of C a dose of  10 mg.kg-1 of the fish© Penyakit Motil Aeromonas Septicaemia (MAS) yang menyerang ikan nila mengalami peningkatan selama beberapa tahun terakhir sebagai konsekuensi dari kegiatan akuakultur intensif, yang menyebabkan kerugian dalam industri budidaya. Agen utama penyebab penyakit MAS adalah Aeromonas hydrophila. Untuk mengendalikan penyakit tersebut dapat dilakukan dengan pemberian β-glukan. Sebagai imunostimulan, β-glukan juga dapat  meningkatkan resistensi pada ikan nila yang dibudidayakan. Pengkajian mengenai pemanfaatan β-glukan yang diekstrak dari ragi roti Saccharomyces cerevisiae dimaksudkan untuk menguji sistem imun non spesifik ikan nila yang diuji tantang dengan bakteri Aeromonas hydrophila. Metode yang digunakan yaitu metode eksperimen dengan rancangan acak lengkap yang terdiri dari empat perlakuan dan tiga ulangan. Dosis β-glukan  yang digunakan sebagai perlakuan sebesar 0 mg.kg-1 ikan (Kontrol), 5 mg.kg-1 ikan (B), 10 mg.kg-1 ikan (C) dan 20 mg.kg-1 ikan (D), masing-masing perlakuan diinjeksi sebanyak 3 kali dengan interval waktu 3 hari selama 9 hari, volume injeksi 0,5 mL/ekor ikan dan pengamatan resistensi selama tujuh hari. Hasil penelitian menunjukkan perbedaan jumlah β-glukan dan frekuensi pemberian yang diinjeksikan memberikan pengaruh nyata terhadap total leukosit, aktivitas fagositosis dan resistensi. Total leukosit, aktivitas fagositosis dan resistensi terbaik dicapai pada perlakuan C dengan dosis 10 mg.kg-1 ikan©

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