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2.3. Activity test for immobilized
2.3. Activity test for immobilized PLA1
Enzymatic modification of PC was evaluated by performing activity tests. A 1.2 g sample of highly enriched n3 PUFA from fish oil (8 mol) and 2.8 g of PC (1 mol) were placed in a 50 mL waterjacketed glass vessel, mixed at 250 rpm and preheated to 55 C. Then, 0.5 g of the enzyme (10% of total substrate weight) was added. Samples (50 mg) of the product mixture were taken at regular intervals, dissolved in chloroform, and applied to silica-coated preparative TLC plates and developed to obtain the PC fraction. After saponification and methylation, the resulting fatty acid methyl esters (FAMEs) were subjected to gas chromatographic analysis. The apparent activity was defined as the initial reaction rate divided by the amount of immobilized enzyme. The specific activity was defined as the initial reaction rate divided by the amount of
protein. The acidolysis reaction involves concurrent decomposition of PC and addition of fatty acid to the molecule. Since esterification of fatty acid should be preceded by hydrolysis of PC, hydrolytic activity of the enzyme was used to measure the initial reaction rate. The apparent activity and the specific activity were calculated as follows:
Enzymatic modification of PC was evaluated by performing activity tests. A 1.2 g sample of highly enriched n3 PUFA from fish oil (8 mol) and 2.8 g of PC (1 mol) were placed in a 50 mL waterjacketed glass vessel, mixed at 250 rpm and preheated to 55 C. Then, 0.5 g of the enzyme (10% of total substrate weight) was added. Samples (50 mg) of the product mixture were taken at regular intervals, dissolved in chloroform, and applied to silica-coated preparative TLC plates and developed to obtain the PC fraction. After saponification and methylation, the resulting fatty acid methyl esters (FAMEs) were subjected to gas chromatographic analysis. The apparent activity was defined as the initial reaction rate divided by the amount of immobilized enzyme. The specific activity was defined as the initial reaction rate divided by the amount of
protein. The acidolysis reaction involves concurrent decomposition of PC and addition of fatty acid to the molecule. Since esterification of fatty acid should be preceded by hydrolysis of PC, hydrolytic activity of the enzyme was used to measure the initial reaction rate. The apparent activity and the specific activity were calculated as follows:
0/5000
2.3. Activity test for immobilized PLA1
Enzymatic modification of PC was evaluated by performing activity tests. A 1.2 g sample of highly enriched n3 PUFA from fish oil (8 mol) and 2.8 g of PC (1 mol) were placed in a 50 mL waterjacketed glass vessel, mixed at 250 rpm and preheated to 55 C. Then, 0.5 g of the enzyme (10% of total substrate weight) was added. Samples (50 mg) of the product mixture were taken at regular intervals, dissolved in chloroform, and applied to silica-coated preparative TLC plates and developed to obtain the PC fraction. After saponification and methylation, the resulting fatty acid methyl esters (FAMEs) were subjected to gas chromatographic analysis. The apparent activity was defined as the initial reaction rate divided by the amount of immobilized enzyme. The specific activity was defined as the initial reaction rate divided by the amount of
protein. The acidolysis reaction involves concurrent decomposition of PC and addition of fatty acid to the molecule. Since esterification of fatty acid should be preceded by hydrolysis of PC, hydrolytic activity of the enzyme was used to measure the initial reaction rate. The apparent activity and the specific activity were calculated as follows:
Enzymatic modification of PC was evaluated by performing activity tests. A 1.2 g sample of highly enriched n3 PUFA from fish oil (8 mol) and 2.8 g of PC (1 mol) were placed in a 50 mL waterjacketed glass vessel, mixed at 250 rpm and preheated to 55 C. Then, 0.5 g of the enzyme (10% of total substrate weight) was added. Samples (50 mg) of the product mixture were taken at regular intervals, dissolved in chloroform, and applied to silica-coated preparative TLC plates and developed to obtain the PC fraction. After saponification and methylation, the resulting fatty acid methyl esters (FAMEs) were subjected to gas chromatographic analysis. The apparent activity was defined as the initial reaction rate divided by the amount of immobilized enzyme. The specific activity was defined as the initial reaction rate divided by the amount of
protein. The acidolysis reaction involves concurrent decomposition of PC and addition of fatty acid to the molecule. Since esterification of fatty acid should be preceded by hydrolysis of PC, hydrolytic activity of the enzyme was used to measure the initial reaction rate. The apparent activity and the specific activity were calculated as follows:
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2.3. Uji aktivitas untuk bergerak PLA1
enzimatik modi fi kasi dari PC dievaluasi dengan melakukan tes aktivitas. Sebuah 1,2 g sampel yang sangat diperkaya n? 3 PUFA dari minyak ikan (8 mol) dan 2,8 g PC (1 mol) ditempatkan dalam bejana kaca waterjacketed 50 mL, dicampur pada 250 rpm dan dipanaskan sampai 55 ° C. Kemudian, 0,5 g enzim (10% dari total substrat berat) ditambahkan. Sampel (50 mg) dari campuran produk diambil secara berkala, dilarutkan dalam kloroform, dan diterapkan untuk preparatif pelat TLC berlapis-silika dan dikembangkan untuk memperoleh fraksi PC. Setelah saponifikasi fi kasi dan metilasi, ester metil asam lemak yang dihasilkan (James) menjadi sasaran analisis kromatografi gas. Kegiatan yang jelas adalah didefinisikan sebagai laju reaksi awal dibagi dengan jumlah enzim amobil. The spesifik Kegiatan fi c adalah didefinisikan sebagai laju reaksi awal dibagi dengan jumlah
protein. Reaksi acidolysis melibatkan dekomposisi bersamaan PC dan penambahan asam lemak ke dalam molekul. Sejak Esteri fi kasi asam lemak harus didahului oleh hidrolisis PC, aktivitas hidrolisis enzim digunakan untuk mengukur laju reaksi awal. Kegiatan jelas dan spesifik aktivitas fi c dihitung sebagai berikut:
enzimatik modi fi kasi dari PC dievaluasi dengan melakukan tes aktivitas. Sebuah 1,2 g sampel yang sangat diperkaya n? 3 PUFA dari minyak ikan (8 mol) dan 2,8 g PC (1 mol) ditempatkan dalam bejana kaca waterjacketed 50 mL, dicampur pada 250 rpm dan dipanaskan sampai 55 ° C. Kemudian, 0,5 g enzim (10% dari total substrat berat) ditambahkan. Sampel (50 mg) dari campuran produk diambil secara berkala, dilarutkan dalam kloroform, dan diterapkan untuk preparatif pelat TLC berlapis-silika dan dikembangkan untuk memperoleh fraksi PC. Setelah saponifikasi fi kasi dan metilasi, ester metil asam lemak yang dihasilkan (James) menjadi sasaran analisis kromatografi gas. Kegiatan yang jelas adalah didefinisikan sebagai laju reaksi awal dibagi dengan jumlah enzim amobil. The spesifik Kegiatan fi c adalah didefinisikan sebagai laju reaksi awal dibagi dengan jumlah
protein. Reaksi acidolysis melibatkan dekomposisi bersamaan PC dan penambahan asam lemak ke dalam molekul. Sejak Esteri fi kasi asam lemak harus didahului oleh hidrolisis PC, aktivitas hidrolisis enzim digunakan untuk mengukur laju reaksi awal. Kegiatan jelas dan spesifik aktivitas fi c dihitung sebagai berikut:
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