2.2. Immobilization of PLA1For the

2.2. Immobilization of PLA1
For the immobilization of PLA1, a free type PLA1 was mixed with sodium phosphate buffer solution (50 mM, pH 7.5) at various ratios. The enzyme suspension was stirred at 300 rpm for 30 min
and its soluble protein concentration was determined according to Lowry, Rosebrough, Farr, and Randall (1951) using bovine serum albumin as the standard. For hydrophobic carriers, one gram of each carrier was wetted with 20 mL of ethanol for 2 h until the carrier sedimented. After decanting ethanol, the carrier was washed with 40 mL sodium phosphate buffer and subsequently placed in a flask with 10 mL of enzyme suspension. Immobilization was performed in a water bath shaker operating at 200 rpm and 30 C. After shaking for 16 h, the suspension was filtered to recover the resulting immobilized enzyme preparation, which was then rinsed with 40 mL of sodium phosphate buffer and then dried at 45 Cina vacuum oven overnight, and stored at 4 C until use. The fixation level (wt%) and the amount of protein bound in immobilized enzyme (mg/g) were estimated by subtracting the protein remaining in the enzyme suspension after immobilization compared with the initial protein concentration. Fixation level (%) and amount of protein adsorbed onto the carrier (mg/g) were calculated by following equations:

Hasil (Bahasa Indonesia) 2:

2.2. Imobilisasi PLA1
Untuk imobilisasi PLA1, jenis PLA1 gratis dicampur dengan penyangga sodium fosfat solusi (50 mM, pH 7,5) pada berbagai rasio. Suspensi enzim diaduk pada 300 rpm selama 30 menit
dan konsentrasi protein terlarut yang ditentukan menurut Lowry, Rosebrough, Farr, dan Randall (1951) menggunakan serum albumin sapi sebagai standar. Untuk operator hidrofobik, satu gram masing-masing operator yang dibasahi dengan 20 ml etanol selama 2 jam sampai pembawa mengendap. Setelah decanting etanol, pembawa dicuci dengan 40 mL dapar natrium fosfat dan kemudian ditempatkan dalam sebuah fl meminta dengan 10 mL suspensi enzim. Imobilisasi dilakukan dalam shaker bath air beroperasi pada 200 rpm dan 30? C. Setelah gemetar selama 16 jam, suspensi itu disaring untuk memulihkan persiapan yang dihasilkan amobil enzim, yang kemudian dibilas dengan 40 ml buffer fosfat natrium dan kemudian dikeringkan pada 45? Cina vakum oven semalam, dan disimpan pada 4 ° C sampai digunakan. Tingkat fi xation (wt%) dan jumlah protein terikat dalam enzim amobil (mg / g) diperkirakan dengan mengurangi protein yang tersisa di suspensi enzim setelah imobilisasi dibandingkan dengan konsentrasi protein awal. Tingkat fiksasi (%) dan jumlah protein teradsorbsi ke pembawa (mg / g) dihitung dengan mengikuti persamaan: