Isolation, identification and mutants generation of isolate 04 for its improved bioconversion of ferulic acid to vanillin
DOI:
https://doi.org/10.71336/jabs.1381Keywords:
Lysinibacillus macroides, Biotransformation, UV-spectrophotometry, HPLC, Physical mutagensAbstract
Vanillin is widely used as a food flavouring, scent ingredient in cosmetics and perfumes, as an intermediate in agrochemicals & medicines and also as a nutraceutical. Serial dilution of sediment sample yielded 32 strains, among which three isolates A2, A7, and 04 reported here were characterized at molecular level by 16S rDNA sequencing. They were identified and similar to Pseudomonas aeruginosa (94.97%), Acinetobacter baumannii (99.93%) and Lysinibacillus macroides (98.67%) respectively based on its 16S rDNA sequences. Among these three strains, isolate 04 was with high ferulic acid (FA) to vanillin bioconversion activity. Vanillin concentration was determined by the Thiobarbituric acid (TBA) method and analysed by UV-spectrophotometry, which showed 11.63 ± 0.05 mg/ml vanillin after optimization studies. HPLC with UV detector was also used for the characterization of the vanillin formed, which showed peak at 8.793 rt. To improve FA to vanillin bioconversion, physical mutagenesis was performed on isolate 04, which generated three mutants (1, 2, and 3). These three mutants were subject to FA to vanillin biotransformation study. TBA method and UV spectrophotometric analysis showed a higher conversion by mutant 2 than other two mutants, 1 and 3. Mutant 2 formed almost two-fold vanillin i.e. 19.78 ± 0.05 mg/ml when compared to its wild strain, isolate 04. Hence, this mutant may be further subjected to optimization studies and exploited for industrial biotransformation process.
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