Isolation, Screening, and Molecular Identification of Pectinase Producing Microorganism

Abstract - Pectinase is one of the most popular industrial enzymes, utilized in the major processes of clarification of fruit juices, textile processing and biofuel and food. Novel pectinase producing bacteria could be isolated using pectin-rich agro-industrial wastes like fruit and vegetable peels, which are cheap waste. Bacterial species were isolated from fruit and vegetable waste samples collected from Haldwani, Nainital district, India and screened for their pectinase activity by a pectin-agar clearance-zone assay. Eight isolates were cultured, of which six were pectinase positive. Isolate B1B that formed the largest hydrolysis zone (18 mm) and exhibited the highest pectinase activity (157 IU/GDS) was chosen for molecular characterization. After the 16S rRNA gene was amplified and sequenced, isolate B1B was determined to be Bacillus cereus strain SH-02 by BLAST search and phylogenetic tree construction. This strain was closely linked to the reference strains of B. cereus HB45, EM10, and Y3. The results presented here show that Bacillus cereus SH-02 is a new bacterium for use in industry as a pectinase producer and can be used as a validated strain in downstream fermentation and optimization studies.