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) 1:

2.2. Imobilisasi atas PLA1Untuk Imobilisasi PLA1, sejenis gratis PLA1 dicampur dengan larutan penyangga natrium fosfat (50 mM, pH 7.5) di berbagai rasio. Suspensi enzim diaduk pada 300 rpm selama 30menitdan konsentrasi protein larut ditentukan menurut Lowry, Rosebrough, Tintin dan Randall (1951) menggunakan sapi albumin serum sebagai standar. Untuk operator hidrofobik, satu gram masing-masing pembawa adalah dibasahi dengan 20 mL etanol untuk 2 h hingga pembawa sedimented. Setelah decanting etanol, pembawa dicuci dengan 40 mL natrium fosfat buffer dan kemudian ditempatkan dalam flask dengan 10 mL suspensi enzim. Imobilisasi dilakukan dalam shaker mandi air beroperasi pada 200 rpm dan 30 C. Setelah gemetar untuk 16 h, suspensi adalah filtered untuk memulihkan yang dihasilkan bergerak enzim persiapan, yang kemudian dibilas dengan 40 mL buffer natrium fosfat dan kemudian kering di oven vakum Cina 45 semalam, dan disimpan di 4 C sampai digunakan. Tingkat fixation (wt %) dan jumlah protein yang terikat dalam bergerak enzim (mg/g) diperkirakan dengan mengurangi protein yang tersisa dalam suspensi enzim setelah Imobilisasi dibandingkan dengan konsentrasi protein awal. Fiksasi tingkat (%) dan jumlah protein adsorbed ke pembawa (mg/g) dihitung oleh persamaan berikut: