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Objective: This study aimed to obtain glucose from α-cellulose of water hyacinth through enzymatic hydrolysis.

Method: The cellulase activity was screened by the formation of clear zone in CMC agar medium and DNS-reducing sugar method. α-Cellulose of water hyacinth was hydrolyzed using crude cellulase enzyme extract of Chaetomium globosum. Glucose identified by FTIR spectrophotometer and qualitative analysis test using Fehling’s test method.

Results: Crude enzyme from Chaetomium globosum has high activity of cellulase with a clear zone diameter of 45.00 mm. Crude enzyme from Chaetomium globosum also showed high activity of cellulase with measurement by sugar reduction method which produced a glucose concentration of 215.617 μg/ml. The optimum conditions of enzymatic hydrolysis to produce glucose from α-cellulose water hyacinth powder were at 50°C, pH 5 and using an enzyme concentration of 2% for 48 hours, 160 rpm. The average glucose concentration obtained from α-cellulose water hyacinth powder in optimum conditions of enzymatic hydrolysis was 214.450 μg/ml with glucose yield 0.429%.

Conclusion: The optimum condition of enzymatic hydrolysis was reached at 50°C, pH 5, with the addition of 2% enzyme concentration, 48 hours,160 rpm. The results of identification with FTIR showed that the spectrum of glucose produced has similarities to the spectrum of glucose standard, especially in the fingerprint region. The identification by the Fehling’s test showed positive results for both glucose standard and solution test from hydrolysis results.