Enzymatic determination of the degree of chitosan deacetylation by family 18 chitinase
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Abstract
The degree of chitosan deacetylation is the ratio of D-glucosamine (GlcN) units to the sum of N-acetyl-D-glucosamine (GlcNAc) and GlcN. An enzymatic method was developed to determine the percent degree of chitosan deacetylation (%DD) samples. A completely hydrolyzed chitosan solution was obtained by using crude chitinase from Bacillus licheniformis SK-1. The optimum condition was as follows: 20 mU of crude chitinase per milligram of chitosan in 0.1 M acetate buffer (pH 4.5) at 40 °C for 8 h. The %DD was determined by generating a standard calibration curve. The total reducing sugar (TRS) and the %DD of standard chitosan samples (ranging from 43% to 100%) were determined by using nuclear magnetic resonance and colloidal titration methods. The standard calibration curve for determining the %DD was obtained with a linear correlation: %DD = − 21.479 (TRS) + 102.45, r2 = 0.9933. The enzymatic method showed reproducible results and did not require special instruments.
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