Characterization of vitamin B12 in Dunaliella salina

Vitamin B12 is one of nature's complex metabolite which is industrially produced using certain bacteria. Algae could be an alternative source of vitamin B12 and in this study, vitamin B12 from a halotolerant green alga, Dunaliella salina V-101 was purified and characterized. The extract of Dunaliella was purified by passing through Amberlite XAD-2 and EASI-extract vitamin B12 immunoaffinity column. The total vitamin B12 content in purified sample fractions was 42 ± 2 μg/100 g dry weight as determined by the chemiluminescence method which was almost close to 49 ± 2 μg/100 g dry weight as estimated by microbiological method. Further quantification of total vitamin B12 using gold nanoparticle (AUNPs) based aptamer showed 40 ± 0.8/100 g dry weight. There was a good correlation among all the methods of quantification. Adenosylcobalamin, a form of vitamin B12 which is a cofactor for methylmalonyl CoA mutase was identified by HPLC. Upon quantification, Dunaliella was found to contain 34 ± 4 μg of adenosylcobalamin for 100 g dry biomass. Authenticity of adenosylcobalmin was confirmed by tandem mass spectrometry (MS/MS), selected ion recording (SIR) and multiple reaction monitoring (MRM) studies.