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Title: Total Protein Analysis in Algae via Bulk Amino Acid Detection: Optimization of Amino Acid Derivatization after Hydrolysis with O-Phthalaldehyde 3-Mercaptopropionic Acid (OPA-3MPA)

Abstract

The analysis of protein in algal biomass is one of the most critical areas of commercial development of algae characterization for nutritional or other high value applications. A new rapid and accurate method is required that can be widely implemented and that is free from interferences from the complex algal biomass matrix. We developed a simple spectrophotometric method for primary amino acid quantification bulk measurement in an acid hydrolyzed algal biomass preparation, as an alternative to the more labor-intensive amino HPLC acid analysis or less specific nitrogen-to-protein conversion. We have validated an O-phthalaldehyde (OPA)-based derivatization method, showing accurate and linear quantification for standard reference amino acids as well as mixtures, mimicking the amino acid complexity found in algal biomass. The presence of interferences that may be derived from the complex biomass biochemical composition was tested during the method validation phase. We document the application of a novel method of OPA derivatization with 3-mercaptopropionic acid (3MPA) to determine the total amino acid content of harvested algal biomass collected from different, controlled cultivation conditions and demonstrated a within 10% accuracy against a reference measurement of amino acid content in at least 4 species and 10 algal biomass samples, across early, mid, andmore » late-stages of cultivation.« less

Authors:
 [1]; ORCiD logo [1]
  1. National Renewable Energy Lab. (NREL), Golden, CO (United States)
Publication Date:
Research Org.:
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Sponsoring Org.:
USDOE Office of Energy Efficiency and Renewable Energy (EERE), Sustainable Transportation Office. Bioenergy Technologies Office
OSTI Identifier:
1514837
Report Number(s):
NREL/JA-5100-73803
Journal ID: ISSN 0021-8561
Grant/Contract Number:  
AC36-08GO28308
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Journal of Agricultural and Food Chemistry
Additional Journal Information:
Journal Volume: 67; Journal Issue: 19; Journal ID: ISSN 0021-8561
Publisher:
American Chemical Society
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; algae; amino acid analysis; O-phthalaldehyde; OPA; spectrophotometry

Citation Formats

Cuchiaro, Joshua H., and Laurens, Lieve M. Total Protein Analysis in Algae via Bulk Amino Acid Detection: Optimization of Amino Acid Derivatization after Hydrolysis with O-Phthalaldehyde 3-Mercaptopropionic Acid (OPA-3MPA). United States: N. p., 2019. Web. doi:10.1021/acs.jafc.9b00884.
Cuchiaro, Joshua H., & Laurens, Lieve M. Total Protein Analysis in Algae via Bulk Amino Acid Detection: Optimization of Amino Acid Derivatization after Hydrolysis with O-Phthalaldehyde 3-Mercaptopropionic Acid (OPA-3MPA). United States. https://doi.org/10.1021/acs.jafc.9b00884
Cuchiaro, Joshua H., and Laurens, Lieve M. 2019. "Total Protein Analysis in Algae via Bulk Amino Acid Detection: Optimization of Amino Acid Derivatization after Hydrolysis with O-Phthalaldehyde 3-Mercaptopropionic Acid (OPA-3MPA)". United States. https://doi.org/10.1021/acs.jafc.9b00884. https://www.osti.gov/servlets/purl/1514837.
@article{osti_1514837,
title = {Total Protein Analysis in Algae via Bulk Amino Acid Detection: Optimization of Amino Acid Derivatization after Hydrolysis with O-Phthalaldehyde 3-Mercaptopropionic Acid (OPA-3MPA)},
author = {Cuchiaro, Joshua H. and Laurens, Lieve M.},
abstractNote = {The analysis of protein in algal biomass is one of the most critical areas of commercial development of algae characterization for nutritional or other high value applications. A new rapid and accurate method is required that can be widely implemented and that is free from interferences from the complex algal biomass matrix. We developed a simple spectrophotometric method for primary amino acid quantification bulk measurement in an acid hydrolyzed algal biomass preparation, as an alternative to the more labor-intensive amino HPLC acid analysis or less specific nitrogen-to-protein conversion. We have validated an O-phthalaldehyde (OPA)-based derivatization method, showing accurate and linear quantification for standard reference amino acids as well as mixtures, mimicking the amino acid complexity found in algal biomass. The presence of interferences that may be derived from the complex biomass biochemical composition was tested during the method validation phase. We document the application of a novel method of OPA derivatization with 3-mercaptopropionic acid (3MPA) to determine the total amino acid content of harvested algal biomass collected from different, controlled cultivation conditions and demonstrated a within 10% accuracy against a reference measurement of amino acid content in at least 4 species and 10 algal biomass samples, across early, mid, and late-stages of cultivation.},
doi = {10.1021/acs.jafc.9b00884},
url = {https://www.osti.gov/biblio/1514837}, journal = {Journal of Agricultural and Food Chemistry},
issn = {0021-8561},
number = 19,
volume = 67,
place = {United States},
year = {Wed Apr 24 00:00:00 EDT 2019},
month = {Wed Apr 24 00:00:00 EDT 2019}
}

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Cited by: 6 works
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