OPTIMIZATION OF LACCASE IMMOBILIZATION ON CHITOSAN BEADS USING THE CROSS-LINKING METHOD
DOI:
https://doi.org/10.20414/spin.v7i1.11828Keywords:
chitosan, cross-linking, glutaraldehyde, immobilization, laccaseAbstract
Limbah industri tekstil merupakan salah satu masalah utama lingkungan saat ini. Limbah tersebut mengandung residu pewarna dan berbagai senyawa kompleks. Lakase menjadi salah satu pilihan untuk degradasi biokatalitik zat warna karena biodegradabilitas dan biokompatibilitasnya. Pemanfaatan lakase untuk detoksifikasi polutan secara luas menjadi terbatas karena kerentanannya terhadap perubahan kondisi operasional. Untuk mengatasi masalah tersebut, lakase diimobilisasi pada kitosan dengan glutaraldehida sebagai agen pengikat silang. Penelitian ini dilakukan untuk menentukan kondisi optimum metode ikat silang dan imobilisasi lakase pada manik-manik kitosan. Pada penelitian ini, lakase dari Aspergillus sp. diimobilisasi pada butiran kitosan yang berikatan silang dengan glutaraldehida. Butiran kitosan dibuat dengan menggunakan kitosan 2% (b/v) dan diikat silang dengan glutaraldehida. Selanjutnya, butiran yang sudah teraktivasi glutaraldehida diinkubasi dengan lakase. Lakase yang terimobilisasi pada butiran kitosan diuji kadar protein dan aktivitasnya untuk memperoleh nilai efisiensi imobilisasi. Hasil penelitian menunjukkan bahwa konsentrasi glutaraldehida optimum adalah 0,8% dengan waktu ikat silang selama 6 jam, dosis lakase 0,4 mg/mL dan waktu imobilisasi selama 4 jam. Dari kondisi ini, didapatkan efisiensi imobilisasi sebesar 16,51% dengan aktivitas lakase sebesar 24,58 U/g. Berdasarkan karakterisasi gugus fungsi manik-manik kitosan terimobilisasi lakase, adanya puncak untuk gugus fungsi C=N mengkonfirmasi bahwa lakase telah terimobilisasi pada kitosan secara kovalen.
The textile industry is one of the significant environmental problems. The waste contains dye residues and various complex compounds. Laccase is a viable option for biocatalytic degradation due to its biodegradability and biocompatibility. The widespread use of laccase for pollutant detoxification is limited due to its susceptibility to changes in operational conditions. To overcome this problem, laccase was immobilized on chitosan with glutaraldehyde as a cross-linking agent. This research was conducted to determine the optimum conditions for the cross-linking method and laccase immobilization on chitosan beads. In this study, chitosan beads were developed using 2% (w/v) chitosan and cross-linked with glutaraldehyde. The activated chitosan beads were then incubated with laccase. The immobilized laccase was tested for its protein content and activity to obtain an immobilization efficiency value. The results showed that the optimum glutaraldehyde concentration was 0.8% with a cross-linking time of 6 hours, a laccase dose of 0,4 mg/mL, and an immobilization time of 4 hours. From these conditions, an immobilization efficiency of 16.51% was obtained with an enzyme activity of 24.58 U/g. Based on the functional group characterization of laccase-immobilized chitosan beads, the presence of the C=N functional group peak confirmed that laccase had been covalently immobilized on chitosan.
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