Protein Engineering of Mung Bean (Vigna radiata (L.) Wilczek) 8Sα Globulin with Lactostatin

Issue Date

12-2020

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Protein Engineering of Mung Bean (Vigna radiata (L.) Wilczek) 8Sα Globulin with Lactostatin

by Ma. Carla Gamis 1,2,*, Lawrence Yves Uy 3,4, Antonio Laurena 1,5, Wilma Hurtada 1 and Mary Ann Torio 1,3 1 Graduate School, University of the Philippines Los Baños, Jose R. Velasco Ave., Los Baños, Laguna 4031, Philippines 2 National Institute of Molecular Biology and Biotechnology, University of the Philippines Diliman, Quezon City 1101, Philippines 3 Institute of Chemistry, University of the Philippines Los Baños, Los Baños, Laguna 4031, Philippines 4 2nd Level, Science Heritage Building, Sibol St. DOST Compound, General Santos Avenue, Bicutan, Taguig City 1631, Philippines 5 Philippine Genome Center-Program for Agriculture, Livestock, Forestry and Fisheries, Office of the Vice Chancellor for Research and Extension, UPLB, Los Baños, Laguna 4031, Philippines * Author to whom correspondence should be addressed. Appl. Sci. 2020, 10(24), 8787; https://doi.org/10.3390/app10248787 Submission received: 19 September 2020 / Revised: 1 November 2020 / Accepted: 3 November 2020 / Published: 8 December 2020 (This article belongs to the Special Issue Bioactive Peptides from Foods) Downloadkeyboard_arrow_down Browse Figures Versions Notes

Abstract

Mung bean is a well-known good source of protein. To increase its bioactivity, economic value, and nutritional content as a functional food and food additive, lactostatin (IIAEK), a cholesterol-lowering bioactive peptide, was engineered into mung bean 8Sα globulin, a major storage protein. The results showed that the mutated 8Sα globulin has a significant bile acid binding capacity (cholesterol-lowering activity) up to 47.25%. Moreover, superimposed mutant (Mut2) and wild-type (Wt) 3D protein structures showed a 93–97% identity, indicating that the mutant proteins are stable. Ultra-performance liquid chromatography(UPLC)-based assay showed similar retention time for wild-type and mutant protein samples. Both IIAEK peptide standard and Mut2 digest had comparable baseline peaks corresponding to the same molecular size based on the liquid chromatography-mass spectrometry (LC-MS) data. A 573.36-Da mass spectrum was seen in Mut2, which indicates that Mut2 8Sα globulin has been successfully mutated and digested to release the bioactive peptide, IIAEK. In vitro bile acid binding capacity showed that the 6-h Wt and 12-h engineered protein (Mut2) digests had the highest activity. Lastly, potential allergenicity was checked in the Allergen Database for Food Safety (ADFS) and the AllerBase database, and the IIAEK peptide matched the Bos d 5 epitopes. This study provides a strong foundation and basis for mung bean nutrition improvement, development of cholesterol-lowering food supplements, and protein engineering of other food proteins. Keywords: lactostatin; mung bean; protein engineering; bioactive peptide

Source or Periodical Title

The 5th International Symposium of Frontiers in Molecular Science

Document Type

Article

Language

English

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