RESEARCH ARTICLE


Microrespirometric Validation of a Two-stage Process for Polyhydroxyalkanoates Production from Peanut Oil and Propionate with Cupriavidus necator



Irazú Pérez-Camacho1, Norberto Chavarría-Hernández1, Ma del Rocío López-Cuellar1, Claudia Guerrero-Barajas2, Alberto Ordaz3, *
1 Cuerpo Académico de Biotecnología Agroalimentaria, Instituto de Ciencias Agropecuarias, Universidad Autónoma del Estado de Hidalgo, Av. Universidad km 1, Rancho Universitario, C.P. 43600, Tulancingo de Bravo, Hidalgo, Mexico
2 Laboratorio de Biotecnología Ambiental, Departamento de Bioprocesos, Unidad Profesional Interdisciplinaria de Biotecnología. Instituto Politécnico Nacional. Av. Acueducto s/n Col. Barrio la Laguna Ticomán, Mexico City, 07340, Mexico
3 Departamento de Bioingeniería, Escuela de Ingeniería y Ciencias, Tecnológico de Monterrey, Campus Estado de México. Carretera Lago de Guadalupe Km 3.5, Margarita Maza de Juárez, Atizapán de Zaragoza, Estado de México, México


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Creative Commons License
© 2024 The Author(s). Published by Bentham Open.

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: https://creativecommons.org/licenses/by/4.0/legalcode. This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

* Address correspondence to this author at the Departamento de Bioingeniería, Escuela de Ingeniería y Ciencias, Tecnológico de Monterrey, Campus Estado de México. Carretera Lago de Guadalupe Km 3.5, Margarita Maza de Juárez, Atizapán de Zaragoza, Estado de México, México; E-mail: alberto.ordaz@tec.mx


Abstract

Introduction

Poly(3-hydroxyalkanoates) (PHA) is a biodegradable polyester synthesized by various bacteria, including Cupriavidus necator. The composition of PHA is influenced by the type of microorganism, cultivation conditions, and carbon substrate. The selection of a carbon source is critical for PHA production, significantly impacting process costs. Therefore, the common goal is scalability, with the design and optimization of PHA production relying on experimental determination of constitutive parameters.

Methods

In this work, a two-stage process for PHA production was conducted in shaking flasks functioning as mini reactors with the bacteria Cupriavidus necator. During the first stage, fructose was used as the sole carbon source. Once fructose was exhausted, a second stage commenced with a new carbon source, either peanut oil or propionate, to enhance PHA production. Ex-situ pulse respirometry approach was employed during the two-stage process to follow the kinetics of substrate consumption.

Results

The results indicated that the use of peanut oil would be advantageous over propionate, resulting in 12.2% more biomass and 13.9% more PHA.

Conclusion

Additionally, the growth rate was 88.9% higher with peanut oil. The characterization by pulse respirometry applied in microreactors, i.e., micro respirometry, allowed for the performance of up to 216 biological experiments to determine four important kinetic and stoichiometric parameters, namely maximum oxygen uptake (rO2max), substrate affinity constant (KS), growth yield (YX/S), and substrate oxidation yield (YO2/S). The values of these parameters indicated that peanut oil would be the best carbon source to promote PHA production during the second stage. The implementation of the microrespirometry technique during the screening of carbon sources for PHA production provided reliable information within a short period and with significantly less experimental effort.

Keywords: PHA, Respirometry, Microreactors, Kinetic parameters, Peanut oil, Propionate.