Substitute Yeast Extract While Maintaining Performance: Showcase Amorpha‐4,11‐Diene Production

ABSTRACT Yeast extract (YE) is a complex nutritional source associated with high performance on microbial production processes. However, its inherent compositional variability challenges its scalability. While prior efforts have focused on growth‐associated products, the dynamics of growth‐uncoupled...

Full description

Saved in:
Bibliographic Details
Main Authors: Carlos Castillo‐Saldarriaga, Christine N. S. Santos, Stephen Sarria, Parayil K. Ajikumar, Ralf Takors
Format: Article
Language:English
Published: Wiley 2024-11-01
Series:Microbial Biotechnology
Subjects:
Online Access:https://doi.org/10.1111/1751-7915.70056
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850224741312364544
author Carlos Castillo‐Saldarriaga
Christine N. S. Santos
Stephen Sarria
Parayil K. Ajikumar
Ralf Takors
author_facet Carlos Castillo‐Saldarriaga
Christine N. S. Santos
Stephen Sarria
Parayil K. Ajikumar
Ralf Takors
author_sort Carlos Castillo‐Saldarriaga
collection DOAJ
description ABSTRACT Yeast extract (YE) is a complex nutritional source associated with high performance on microbial production processes. However, its inherent compositional variability challenges its scalability. While prior efforts have focused on growth‐associated products, the dynamics of growth‐uncoupled production, which leads to higher production rates and conversion yields, still need to be explored. This production scenario is common in large‐scale applications. This study presents a systematic approach to replace YE for the production of the terpene amorpha‐4,11‐diene in Escherichia coli. Sequential processing was successfully applied to identify glutamic acid, alanine, leucine, valine, isoleucine and glycine as the key amino acids (AAs) under slow‐growth conditions. Thoroughly applying biomass retention as part of sequential processing increased production capacity by 45% using these AAs instead of YE. Further studies, including flux balance analyses, targeted pyruvate as the common AA precursor. The optimized fed‐batch process feeding pyruvate with 0.09 gPyr h−1 enhanced amorpha‐4,11‐diene production by 37%, although adding only 1% carbon via pyruvate. Flux balance analysis revealed the criteria for optimum pyruvate feeding, for example, to prevent succinate secretion and maintain the NADH/NAD+ balance. These findings illustrate the interplay between media composition and metabolic activity and provide a successful guideline for identifying lean, best‐performing media for industrial applications.
format Article
id doaj-art-388e1bbcf1754c809bb68f94e409f427
institution OA Journals
issn 1751-7915
language English
publishDate 2024-11-01
publisher Wiley
record_format Article
series Microbial Biotechnology
spelling doaj-art-388e1bbcf1754c809bb68f94e409f4272025-08-20T02:05:32ZengWileyMicrobial Biotechnology1751-79152024-11-011711n/an/a10.1111/1751-7915.70056Substitute Yeast Extract While Maintaining Performance: Showcase Amorpha‐4,11‐Diene ProductionCarlos Castillo‐Saldarriaga0Christine N. S. Santos1Stephen Sarria2Parayil K. Ajikumar3Ralf Takors4Institute of Biochemical Engineering University of Stuttgart Stuttgart GermanyManusBio Cambridge Massachusetts USAManusBio Cambridge Massachusetts USAManusBio Cambridge Massachusetts USAInstitute of Biochemical Engineering University of Stuttgart Stuttgart GermanyABSTRACT Yeast extract (YE) is a complex nutritional source associated with high performance on microbial production processes. However, its inherent compositional variability challenges its scalability. While prior efforts have focused on growth‐associated products, the dynamics of growth‐uncoupled production, which leads to higher production rates and conversion yields, still need to be explored. This production scenario is common in large‐scale applications. This study presents a systematic approach to replace YE for the production of the terpene amorpha‐4,11‐diene in Escherichia coli. Sequential processing was successfully applied to identify glutamic acid, alanine, leucine, valine, isoleucine and glycine as the key amino acids (AAs) under slow‐growth conditions. Thoroughly applying biomass retention as part of sequential processing increased production capacity by 45% using these AAs instead of YE. Further studies, including flux balance analyses, targeted pyruvate as the common AA precursor. The optimized fed‐batch process feeding pyruvate with 0.09 gPyr h−1 enhanced amorpha‐4,11‐diene production by 37%, although adding only 1% carbon via pyruvate. Flux balance analysis revealed the criteria for optimum pyruvate feeding, for example, to prevent succinate secretion and maintain the NADH/NAD+ balance. These findings illustrate the interplay between media composition and metabolic activity and provide a successful guideline for identifying lean, best‐performing media for industrial applications.https://doi.org/10.1111/1751-7915.70056amino acidscomplex mediaEscherichia colifermentationpyruvateterpenoids
spellingShingle Carlos Castillo‐Saldarriaga
Christine N. S. Santos
Stephen Sarria
Parayil K. Ajikumar
Ralf Takors
Substitute Yeast Extract While Maintaining Performance: Showcase Amorpha‐4,11‐Diene Production
Microbial Biotechnology
amino acids
complex media
Escherichia coli
fermentation
pyruvate
terpenoids
title Substitute Yeast Extract While Maintaining Performance: Showcase Amorpha‐4,11‐Diene Production
title_full Substitute Yeast Extract While Maintaining Performance: Showcase Amorpha‐4,11‐Diene Production
title_fullStr Substitute Yeast Extract While Maintaining Performance: Showcase Amorpha‐4,11‐Diene Production
title_full_unstemmed Substitute Yeast Extract While Maintaining Performance: Showcase Amorpha‐4,11‐Diene Production
title_short Substitute Yeast Extract While Maintaining Performance: Showcase Amorpha‐4,11‐Diene Production
title_sort substitute yeast extract while maintaining performance showcase amorpha 4 11 diene production
topic amino acids
complex media
Escherichia coli
fermentation
pyruvate
terpenoids
url https://doi.org/10.1111/1751-7915.70056
work_keys_str_mv AT carloscastillosaldarriaga substituteyeastextractwhilemaintainingperformanceshowcaseamorpha411dieneproduction
AT christinenssantos substituteyeastextractwhilemaintainingperformanceshowcaseamorpha411dieneproduction
AT stephensarria substituteyeastextractwhilemaintainingperformanceshowcaseamorpha411dieneproduction
AT parayilkajikumar substituteyeastextractwhilemaintainingperformanceshowcaseamorpha411dieneproduction
AT ralftakors substituteyeastextractwhilemaintainingperformanceshowcaseamorpha411dieneproduction