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Twenty three isolates which were collected from soil samples around UTAR, Kampar, Perak and Pantai Bagan Nakhoda Omar, Sabak, Selangor were successfully isolated using chitinase selective medium supplemented with colloidal chitin and bromocresol purple as indicator. Based on their enzyme production, two most potent isolates were selected for identification. The isolates were tentatively identified as Enterobacter sp. and Zymomonas sp. based on their morphological and biochemical test. The preliminary study showed that both Enterobacter sp. and Zymomonas sp. produced the highest chitinase at 48 hours. The process optimization was carried out using one-factor-at-a-time and the parameters involved were the percentage of colloidal chitin, incubation temperature and pH. The optimum parameters for Enterobacter sp. to produce chitinase were 5 % colloidal chitin, at 30 °C and pH 7 with 2.92 U/mL/day of productivity. As for Zymomonas sp., the chitinase productivity (2.54 U/mL/day) was obtained from the optimum conditions i.e. 15 % colloidal chitin at 30 °C and pH 6. citrate utilization, catalase, hydrogen sulfide, indole, triple sugar iron, starch hydrolysis, gelatin hydrolysis, oxidase, mobility, oxidation fermentation, methyl red and Voges Proskauer. E. Optimization of Chitinase Production The optimization was carried based on the one-factor-at-a-time where Lauria Bertaini (LB) broth (g/L: tryptone, 10; yeast extract, 5; NaCl, 10 amended with colloidal chitin 5, pH 7) was used for the chitinase production. To determine the optimum time for chitinase production, the bacteria (10%) was inoculated and incubated at 30 °C, 200 rpm for 7 days. Sampling was carried out every 24 h, where the cultures were centrifuged at 10,000 rpm for 14 min and the supernatant was used for chitinase assay. Effect of different concentration of colloidal chitin (5 – 20 g/L) was carried to determine the best substrate concentration. After that, with the optimized substrate concentration, different incubation temperature (28 – 30 °C) will be investigated and follow by different pH (4 – 7).

Process Optimization on Chitinase Production by Locally Isolated Enterobacter sp. and Zymomonas sp.