Unlocking the Potential of Marine Asparaginase Sources

The marine habitat is a pool of valuable microorganisms with novel metabolites and potential products for industrial applications due to the challenging environment and diversity of living things. It is a proposed source for various commercial enzymes, including the asparaginase enzyme. There is a dearth of marine research conducted off the coast of India, particularly regarding biotechnology and industrial applications. Currently, the available types of L-asparaginase have thermostability and allergy issues. This study isolated and identified microorganisms capable of producing L-asparaginase from Arattuvazhy Golden Beach and Veli Beach's marine water in Thiruvananthapuram, Kerala State. Eleven novel bacteria strains were identified, then identified at the molecular level using suitable primers, followed by sequencing and a phylogenetic tree. Three novel species were selected from the Bacillus genera to clone their asparaginase genes for their novelty. Bacillus zhangzhouensis B29 is not reported for the production of L-asparaginase or any other enzymes; also, L-asparaginase I from B. alititudinis and B. australimaris B28A are not reported. The asparaginase genes (ansA and ansB) were cloned from three species except for ansB from B. australimaris B28A, in which the whole genome sequence was done to retrieve the sequence. Finally, ansB, which is expressed as L-asparaginase II, is identified using the annotated genome sequencing (WGS) of B. australimaris B28A, which is cloned and expressed in the bacterial system with a length of 1149 bp and a homo tetramer size unit of 41 kDa. The SDS-PAGE image and the sequencing verification of the cloned plasmid's open reading frame confirm the gene's in silico analyses. The present study confirms the need for further optimisation of culture conditions for the commercial application of the new L-asparaginase source isolated from India's southern marine zone.