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Characterization of in silico modeled synthetic protein enriched with branched-chain amino acids expressed in Pichia pastoris.

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Relation http://ir.cftri.com/14735/
https://doi.org/10.1016/j.ijbiomac.2020.12.080 0141-8130
 
Title Characterization of in silico modeled synthetic protein enriched with
branched-chain amino acids expressed in Pichia pastoris.
 
Creator Sunil, L.
Prakruthi, Appaiah
Asha, Martin
Prasanna, Vasu
 
Subject 16 Protein Biochemistry
02 Amino acids
 
Description We have designed earlier the 3-dimensional structure of protein enriched with 56% branched-chain amino acids
(BCAA) based on an α-helical coiled-coil structure. The chemically synthesized DNA (BCAA51 gene) was
expressed in Pichia pastoris and confirmed by SDS-PAGE and western blot analysis. In the present study, the purified recombinant protein was characterized using circular dichroism and data revealed that the secondary
structure contained 53.5% α-helix, 3.2% β-strand, and 43.3% turns, which is in concurrence with the overall structure predicted by in silico modeling. The LC-ESI-MS/MS spectra revealed that three peptide masses showed similarity to peptides like EQLTK, LEIVIR, and ILDK, of the modeled BCAA51 protein with the sequence coverage of
~16% from N-terminal region. The N-terminal sequence of the first seven amino acid residues (EQLTKLE) was exactly matching with the in silico designed protein. In vitro digestibility of the protein using SGF and SIF showed the
disappearance of ~11 kDa band and appearance of low molecular weight peptides, which indicated that the protein was easily digestible and non-allergenic, which is the overall objective of this study. Further in vivo digestibility and toxicology studies are required to conclusively utilize this protein as a supplement for the treatment
of chronic liver diseases.
 
Date 2021
 
Type Article
PeerReviewed
 
Format pdf
 
Language en
 
Identifier http://ir.cftri.com/14735/1/International%20Journal%20of%20Biological%20Macromolecules%20168%20%282021%29%20518%E2%80%93525.pdf
Sunil, L. and Prakruthi, Appaiah and Asha, Martin and Prasanna, Vasu (2021) Characterization of in silico modeled synthetic protein enriched with branched-chain amino acids expressed in Pichia pastoris. International Journal of Biological Macromolecules, 168. pp. 518-525. ISSN 0141-8130