Direct Screening and Isolation of Carboxymethyl Cellulase producing Bacilli from Environmental Sources of Khyber Pakhtunkhwa

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Research Paper 01/09/2020
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Direct Screening and Isolation of Carboxymethyl Cellulase producing Bacilli from Environmental Sources of Khyber Pakhtunkhwa

Sahar Nigar, Bashir Ahmad, Sumera Afzal Khan, Ibadullah Jan, Amin Ullah
Int. J. Biosci.17( 3), 34-48, September 2020.
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The bacterial species, which belong to the genus Bacillus, can be economically exploited for the efficient industrial production of carboxymethyl cellulases (CMCases/endo-β-1,4-glucanases) by utilizing the cheap cellulose-based medium. The present study was undertaken to directly screen and isolate CMCase producing bacilli from cellulose associated environmental sources of Khyber Pakhtunkhwa. The screening and isolation of 150 samples were performed by heat shock, serial dilution, and spread plate techniques using Czapek-mineral-salt (CMS) agar medium, Congo red and Gram’s iodine clearing zone assays. The isolates were purified on CMS agar medium and were identified. The CMCase positive efficient isolates were selected by using Congo red and Gram’s iodine assays. The overall population of CMCase positive bacilli varied from 3.2×105±1 to 2.9×106±1 CFU/g in paper-pulp/paper waste (PPW) and organic-fertilizer/manure/animal waste (OMA) respectively. A total of 609 isolates were obtained with an average distribution of 20% in the central districts of the province. The maximum diversity of CMCase positive Bacillus isolates was observed in OMA samples. Out of the 609 isolates, 16 isolates displayed a significant CMCase activity, whereas a high, moderate and lower CMCase activity was exhibited by 100, 325, and 168 isolates, respectively. This study concludes that direct screening and isolation is a good primary screening technique for isolation of industrially important CMCase producing bacilli. Further, it may provide the basis for the utilization of inexpensive cellulose-based medium for the economical production of CMCases and other bio-products on an industrial scale to meet the local and national/international needs of such products.


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