ISOLATION, BIOCHEMICHAL AND MOLECULAR CHARATERIZATION OF PECTINASE ENZYME PRODUCING ACINETOBACTER SP. HSTU-6 BACILLUS SP. HSTU-7 AND HSTU-8 BACTERIA AND ITS APPLICATION TO BIOSCOURING OF CELLULOSIC FIBERS

Abstract

The conventional scouring process involving the harsh environment is slowly being replaced with environment friendly approach using enzymes. These enzymes remove the noncellulosic impurities present in the fabric. Such a process would enhance the absorbency of the fabric without appreciable strength loss and would also help in the proper dyeing and finishing of the fabric. In the present study pectinase enzyme was isolated from cow dung and cow rumen and was optimized under different cultural conditions. Gram staining result isolated from Cow dung of HSTU 6- and HSTU-7 gave positive results, while the isolate HSTU-8 from Rumen content showed negative result. The biochemical tests of Acinetobacter sp.HSTU-6, Bacillus sp. HSTU-7 and HSTU-8 results indicated that Catalase, Oxidase, Methyl red, Maltose, lactose and Dextrose test and selective media Salmonella Agar, Eosinmethyl Agar, Indole, Manintol Salt Agar, and 110 media Agar was found to be positive in this study. And the results of HSTU-6, HSTU-7 and HSTU-8 of Voges-Prokuer (Vp), Citrate, Gelatin mantol salts agar, are found to be negative. The isolate of HSTU6- and HSTU-8 of Triple sugar Iron ager, indicate positive results, while the isolate HSTU-7 indicated negative result, the MIU, results of HSTU-6 and HSTU-7 were positive, while the isolate HSTU-8 indicated negative result, Sucrose fermentation of HSTU-6 and HSTU-8 are positive, while the HSTU-7 was negative. The molecular characterization results were obtained by using 16s rRNA gene sequencing method with the help of the Polymerase chain reaction (PCR) machine and genetic analyzer ABI 3130. From the analysis forward and reverse primers are fully presented in results and discussions. Also the results for Gel Electrophoresis, spectroscopic data, Nucleotide sequences of are given in the result section. HSTU-6 and HSTU-7 Blast search result was obtained, the HSTU-6 and HSTU-7 Phylogenetic tree result are presented; the results of bio-chemical analysis and molecular characterization of pectinase enzyme testify that the enzymes can be used for obscuring of fabrics in large quantity industrially. The result Lignin degradation test and Cross activity test of Acinetobacter sp, HSTU-6 and Bacillus sp, HSTU-7 showed that isolate HSTU-8 strain grow well on try pan blue, Congo red, Avitera blue and bromo phenol blue plates, consequently the clear holo zones were observed. But the isolates HSTU-6, HSTU-7 and HSTU-8 didn‘t grow well on Toluidine blue. The decolorization of methylene blue has been used previously as an indicator of lignin peroxidase enzyme activity. The effectiveness of the dying from all my agricultural products analyzed, the results of Acinetobacter sp. HSTU- 6, Bacillus sp. HSTU-7 and HSTU-8 of the agricultural products mentioned using the pectinase iv enzymes can be used as another alternative source of utilizing for obscuring of cotton fabric for industrial purposes. The Acinetobacter sp.HSTU-6, Bacillus sp. HSTU-7 and HSTU-8 and all results prove that absorbency and more color intensity of the cotton fibers were good enough to compare with untreated control. The results on dyeing were found to be excellent, showing increase dye uptake due to this pretreatment as compared to untreated control. Acinetobacter sp.HSTU-6, Bacillus sp. HSTU-7 and HSTU-8 Color measurment proves the effciency of the strains and visual observation confirms the color difference that exist between the treated samples and untreated sample as shown in tables and figures, Respectivly.