Cellulase Production by Pink Pigmented Facultative Methylotrophic Strains (PPFMs) |
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Authors: | Shanmugam Jayashree Rajendran Lalitha Ponnusamy Vadivukkarasi Yuko Kato Sundaram Seshadri |
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Institution: | (1) Shri AMM Murugappa Chettiar Research Centre (MCRC), Taramani, Chennai, 600113, India;(2) Microbiological and Analytical Group, Food Research Laboratories, Mitsui Norin Co. Ltd, 223-1, Miyahara, Fujieda Shizuoka, 426-0133, Japan; |
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Abstract: | Pink pigmented facultative methylotrophs (PPFM) isolated from water samples of Cooum and Adyar rivers in Chennai and soil
samples of forests located in various districts of Tamil Nadu, India were screened for cellulase production using carboxymethylcellulose
agar (CMC agar) medium. The strains showed wide variations in the production of clearing zones around the colonies on CMC
agar medium flooded with Congo red. CMCase and filter paper assays were used to quantitatively measure the cellulase activity
of 13 PPFM strains. Among the strains, Methylobacterium gregans, MNW 60, MHW 109, MSF 34, and MSF 40 showed cellulolytic activity ranging from 0.73 to 1.16 U mL−1 with wide temperature (35–65°C) and pH (5 to 8) tolerance. SDS-PAGE analysis of the crude enzyme of PPFM strain MNW 60 exhibited
several protein bands, and zymogram analysis revealed two dimeric cellulase bands with molecular mass of ~92 and 42 kDa. Scanning
electron microscopic studies revealed significant morphological differences between the cells grown in normal and CMC amended
medium. The strain MNW 60 was identified as Methylobacterium sp. based on biochemical, physiological, and morphological analyses, and the methylotrophic nature was authenticated by the
presence of mxaF gene, encoding methanol dehydrogenase as a key indicator enzyme of methylotrophs, with 99% similarity to Methylobacterium lusitanum. With the 16S ribosomal RNA sequence showing 97% similarity to M. lusitanum strain MP2, this can be proposed as a novel taxon of the genus Methylobacterium. The study forms the first detailed report on the extracellular cellulase production by pink pigmented Methylobacterium sp., and it is expected that this might be the basis for further studies on cellulase production by PPFMs to explore the
molecular mechanism, strain improvement, and large-scale cellulase production for its application. |
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