The influence of carbon sources and mononucleotides on the production of extracellular alkaline phosphatase by bacillus intermedium

The biosynthesis of extracellular alkaline phosphatase in the streptomycin-resistant strains Bacillus intermedius S3-19 and S7 in the presence in the medium of 5′-nucleoside monophosphates and different sources of carbon-glucose, sodium pyruvate, sodium lactate, or glycerol-was studied. It was established that, in the presence of mononucleotides, the content of extracellular alkaline phosphatase in both strains increased; the maximal effect was caused by 5′-AMP at a concentration of 20 jig/ml. In medium with a low orthophosphate content, where active biosynthesis of alkaline phosphatase occurred, 1 % glucose and 0.5% pyruvate stimulated this process 2.5-4 times, and 2% sodium lactate and sodium pyruvate, on the contrary, inhibited it by 20-40%. Analysis of the dynamics of growth and accumulation of extracellular phosphatase in the presence of different sources of carbon in the medium gives evidence of an interrelationship between the biosynthesis of alkaline phosphatase and carbon metabolism in Bacillus intermedius

A novel Bacillus intermedius extracellular alkaline phosphatase: Isolation, physico-chemical and catalytic characteristics

A new alkaline phosphatase was obtained as homogeneous preparation from culture filtrate of the spore-forming Bacillus intermedius. B. intermedius phosphatase was shown to be monomer with molecular weight of 47 kDa. The enzyme possesses phosphomonoesterase and phosphodiesterase activities and exhibits a broad specificity towards a wide variety of substrates. The purified phosphatase had an optimum temperature of 50°C, optimum pH of 9.5 and was stable until 60°C at pH 8-10. The effect of divalent metal ions and thiol reagents on catalytic activity of the enzyme was studied

Secreted hydrolases from streptomycin-resistant strains of Bacillus intermedius

Alkaline phosphatase and serine proteinase have been isolated from streptomycin-resistant strains of Bacillus intermedius using ion-exchange, affinity and FPLC chromatography. Substrate blotting analysis and electrophoresis revealed two phosphatase forms with molecular weight of 40 and 50 kDa. The pH and temperature optima of phosphatase were at pH 9.5 and 50°C. The enzyme showed a broad substrate specificity. It was suggested that the two forms of phosphatase are the products of processing, in which serine proteinase is the participant. Two proteinase peaks with molecular weights of 29 and 33 kDa were isolated from B. intermedius S7, the first peak having only 5% of the activity of the second peak. The major peak was identical to serine proteinase described earlier. The minor peak was distinct from the major one by the pH-optima. Analysis of inhibitors' effects revealed that the minor peak also corresponded to serine proteinase

Isolation and properties of extracellular alkaline phosphatase from bacillus intermedius

Alkaline phosphatase (APase) was isolated from the culture liquid of the streptomycin-resistant strain of Bacillus intermedius S3-19 and purified as a homogeneous preparation by ion-exchange chromatography and FPLC. Electrophoresis and gel-filtration revealed that the active enzyme is a monomer with molecular weight of 46-47 kD. The enzyme possessed phosphomonoesterase and phosphodiesterase activities with maximal levels at pH 9.5 and 55°C and was stable until 60°C at pH 8.0-10.0. The isolated APase exhibits a broad specificity towards a wide variety of substrates. The effect of divalent metal ions and other reagents on its catalytic activities was studied. It was concluded that alkaline phosphatase of B. intermedius is similar to the secreted alkaline phosphatases from other Bacillus species in its physicochemical and catalytic properties

Secreted hydrolases from streptomycin-resistant strains of Bacillus intermedius

Alkaline phosphatases and serine proteinases have been isolated from the culture liquid of streptomycin-resistant strains of Bacillus intermedius using ion-exchange and affinity chromatography and FPLC. Substrate blotting and electrophoresis revealed two phosphatase forms with molecular masses of 40 and 50 kD. The enzyme had maximal activity at pH 9.5 and 50°C and could cleave the phosphate moiety from a range of substrates. It is suggested that both forms of the phosphatase are products of processing that involves a serine proteinase. Two proteinases, with molecular masses of 29 and 33 kD, were purified to homogeneity from the culture liquid of A. intermedius S7. The protein from the major peak was identical in its properties to an earlier described serine proteinase. The minor peak was 5% of the major one. These enzymes had different pH optima. Inhibitor analysis indicated that the minor peak is also a serine proteinase

Isolation and properties of extracellular alkaline phosphatase from Bacillus intermedim

Alkaline phosphatase (APase) was isolated from the culture liquid of the streptomycin-resistant strain of Bacillus intermedius S3-19 and purified as a homogeneous preparation by ion-exchange chromatography and FPLC. Electrophoresis and gel-nitration revealed that the active enzyme is a monomer with molecular weight of 46-47 kD. The enzyme possessed phosphomonoesterase and phosphodiesterase activities with maximal levels at pH 9.5 and 55°C and was stable until 60°C at pH 8.0-10.0. The isolated APase exhibits a broad specificity towards a wide variety of substrates. The effect of divalent metal ions and other reagents on its catalytic activities was studied. It was concluded that alkaline phosphatase of B. intermedius is similar to the secreted alkaline phosphatases from other Bacillus species in its physicochemical and catalytic properties

The influence of carbon sources and mononucleotides on the production of extracellular alkaline phosphatase by Bacillus intermedium

The biosynthesis of extracellular alkaline phosphatase in the streptomycin-resistant strains Bacillus intermedius S3-19 and S7 in the presence in the medium of 5′-nucleoside monophosphates and different sources of carbon - glucose, sodium pyruvate, sodium lactate, or glycerol-was studied. It was established that, in the presence of mononucleotides, the content of extracellular alkaline phosphatase in both strains increased; the maximal effect was caused by 5′-AMP at a concentration of 20 μg/ml. In medium with a low orthophosphate content, where active biosynthesis of alkaline phosphatase occurred, 1% glucose and 0.5% pyruvate stimulated this process 2.5-4 times, and 2% sodium lactate and sodium pyruvate, on the contrary, inhibited it by 20-40%. Analysis of the dynamics of growth and accumulation of extracellular phosphatase in the presence of different sources of carbon in the medium gives evidence of an interrelationship between the biosynthesis of alkaline phosphatase and carbon metabolism in Bacillus intermedius. © 2000 MAIK "Nauka/Interperiodica"

The influence of carbon sources and mononucleotides on the production of extracellular alkaline phosphatase by bacillus intermedium

The biosynthesis of extracellular alkaline phosphatase in the streptomycin-resistant strains Bacillus intermedius S3-19 and S7 in the presence in the medium of 5′-nucleoside monophosphates and different sources of carbon-glucose, sodium pyruvate, sodium lactate, or glycerol-was studied. It was established that, in the presence of mononucleotides, the content of extracellular alkaline phosphatase in both strains increased; the maximal effect was caused by 5′-AMP at a concentration of 20 jig/ml. In medium with a low orthophosphate content, where active biosynthesis of alkaline phosphatase occurred, 1 % glucose and 0.5% pyruvate stimulated this process 2.5-4 times, and 2% sodium lactate and sodium pyruvate, on the contrary, inhibited it by 20-40%. Analysis of the dynamics of growth and accumulation of extracellular phosphatase in the presence of different sources of carbon in the medium gives evidence of an interrelationship between the biosynthesis of alkaline phosphatase and carbon metabolism in Bacillus intermedius

The influence of carbon sources and mononucleotides on the production of extracellular alkaline phosphatase by bacillus intermedium

The biosynthesis of extracellular alkaline phosphatase in the streptomycin-resistant strains Bacillus intermedius S3-19 and S7 in the presence in the medium of 5′-nucleoside monophosphates and different sources of carbon-glucose, sodium pyruvate, sodium lactate, or glycerol-was studied. It was established that, in the presence of mononucleotides, the content of extracellular alkaline phosphatase in both strains increased; the maximal effect was caused by 5′-AMP at a concentration of 20 jig/ml. In medium with a low orthophosphate content, where active biosynthesis of alkaline phosphatase occurred, 1 % glucose and 0.5% pyruvate stimulated this process 2.5-4 times, and 2% sodium lactate and sodium pyruvate, on the contrary, inhibited it by 20-40%. Analysis of the dynamics of growth and accumulation of extracellular phosphatase in the presence of different sources of carbon in the medium gives evidence of an interrelationship between the biosynthesis of alkaline phosphatase and carbon metabolism in Bacillus intermedius

The influence of carbon sources and mononucleotides on the production of extracellular alkaline phosphatase by Bacillus intermedium

The biosynthesis of extracellular alkaline phosphatase in the streptomycin-resistant strains Bacillus intermedius S3-19 and S7 in the presence in the medium of 5′-nucleoside monophosphates and different sources of carbon - glucose, sodium pyruvate, sodium lactate, or glycerol-was studied. It was established that, in the presence of mononucleotides, the content of extracellular alkaline phosphatase in both strains increased; the maximal effect was caused by 5′-AMP at a concentration of 20 μg/ml. In medium with a low orthophosphate content, where active biosynthesis of alkaline phosphatase occurred, 1% glucose and 0.5% pyruvate stimulated this process 2.5-4 times, and 2% sodium lactate and sodium pyruvate, on the contrary, inhibited it by 20-40%. Analysis of the dynamics of growth and accumulation of extracellular phosphatase in the presence of different sources of carbon in the medium gives evidence of an interrelationship between the biosynthesis of alkaline phosphatase and carbon metabolism in Bacillus intermedius. © 2000 MAIK "Nauka/Interperiodica"