Abstract
Extreme environments are the main source of industrially suitable biocatalysts. The non-cultivable approach of searching enzymes is known to provide ample scope to accomplish novelty for their industrial applications. Lip479 clone out of seven lipase-producing clones obtained from Taptapani hot spring was found to be optimally active at pH 8.0 and temperature 65 °C. The recombinant Lip479 was highly stable in organic solvents, methanol, DMF, DMSO, acetone, and dichloromethane. Lip479 lipase activity was enhanced in the presence of K+, Mn2+, Na+, Zn2+, and Ca2+ except for Fe3+. The ability of Lip479 lipase to act on long carbon chain of 4-nitrophenyl myristate suggests it might be a true lipase. Lip479 clone was found to have ORF of 1251 bp encoding 416 amino acid residues of 42.57 KDa size (theoretically calculated). The presence of conserved motif Ala-His-Ser-Gln-Gly and Zn2+-binding consensus sequence (GAAHAAKH) of the clone assigns the protein to lipase family 1.5. Phylogenetic lineage of the protein sequence of Lip479 was traced to family 1.5 as it was clubbed up with those of reported lipases of the same family. The above biochemical features indicated that Lip479 lipase can be a potential biocatalyst for its use in various industries.
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Present research work received the financial support from the DBT, New Delhi through project grants no: BT/PR7944/BCE/8/1036/2013).
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Fig. S1
Plate showing lipase positive clones. a LB agar containing olive oil and Rhodamine B, b LB agar containing tributyrin (PNG 1002 kb)
Fig. S2
Lip479 clone (NCBI Accession No. KR232658) of 1251 bp length which codes 416 amino acid residues. Signal peptide was marked in red colour outline. Zinc-binding site and conserved motif sequence were marked in brown and blue colour outline respectively. The gene sequence has ATG as start codon (green outline) and TAA as stop codon (black outline). Small alphabets represent nucleotides and block alphabets represent the amino acids (PNG 1772 kb)
Fig. S3
pH stability of Lip479. pH stability was investigated by incubating the crude enzyme at pH 7, 8, 9, 10, and 11 for a period ranging from 0 to 6 h (PNG 131 kb)
Fig. S4
Thermostability of Lip479. Thermostability was measured by incubating the crude enzyme at different temperatures 55, 65, 75, 85, and 95 °C for a period ranging from 0 to 6 h (PNG 137 kb)
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Sahoo, R.K., Das, A., Sahoo, K. et al. Characterization of novel metagenomic–derived lipase from Indian hot spring. Int Microbiol 23, 233–240 (2020). https://doi.org/10.1007/s10123-019-00095-z
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DOI: https://doi.org/10.1007/s10123-019-00095-z