IDENTIFICATION AND PARTIAL CHARACTERIZATION OF CRUDE EXTRACELLULAR ENZYMES FROM BACTERIA ISOLATED FROM SHRIMP WASTE PROCESSING
Abstract: Attention on chitin
degrading enzymes has been growing since their ability to reduce the waste of
shrimp/other crustaceans processing industries and converting them into value
added products such as biologically active chitin and chitosan oligomer. Previous
experiment found that KLU 11.16 isolate was one of the potential bacteria
isolated from shrimp waste producing chitinolytic enzymes including
chitosanases. A study on the identification of KLU 11.16 extracellular crude
enzyme was carried out by cultivating the bacteria on chitin medium. Due to the
wide application of chitosanase, the characterization of the crude chitosanase
was carried out after an identification of the enzymes secreted. Based on
assessment using zymogram technique, this bacteria secreted a mixed
extracellular chitinolytic enzyme and other hydrolytic enzyme. The crude chitinolytic
enzyme degraded 85% deacetylated (DA) better than 100% DA chitosan, and
slightly degraded glycol chitin, indicating that KLU 11.16 secreted
chitosanases and chitinases enzyme. In addition to the chitinolytic enzyme, the
bacteria also secreted protein and carbohydrate degrading enzymes when running
at SDS-PAGE enriched with casein, soluble starch and CMC substrates. Crude
chitosanases enzyme was performed well at pH 6 and temperature of 300 C, and
the activity can be increased by addition of 1 mM Fe 2+ in form of chloride
salt. Addition of detergent, i.e1mM of Triton X-100 and SDS slightly decreased
the activity. Future application of the crude chitosanase from KLU 11.16 was on
producing chitosan derivative such as chitosan oligomer using substrate of 85%
DA chitosan, which is more digestable by other enzymes secreted by KLU 11.16
Keywords: extracellular
enzyme, KLU 11.16, identification
Author: Ekowati Chasanah,
Mahrus Ali, Miftahul Ilmi
Journal Code: jpperikanangg120014
