Wenger, Alexander; Bär, Cornelia; Portmann, Reto; Schmidt, Remo S.; Eugster, Elisabeth; Weisskopf, Laure; Irmler, Stefan (2023). The aminotransferase Aat initiates 3-phenyllactic acid biosynthesis in Pediococcus acidilactici Frontiers in Microbiology, 14 Frontiers Research Foundation 10.3389/fmicb.2023.1150425
|
Text
fmicb-14-1150425.pdf - Published Version Available under License Creative Commons: Attribution (CC-BY). Download (1MB) | Preview |
The function of the aminotransferase Aat (GenBank Protein WP_159211138) from Pediococcus acidilactici FAM 18098 was studied in vivo. For this purpose, the gene was replaced with an erythromycin resistance gene using the temperature-sensitive Escherichia coli-Pediococcus shuttle plasmid pSET4T_Δaat. The knockout was verified by PCR and genome sequencing. Subsequently, the differences between the metabolism of the knockout and of the wild-type strain were investigated by determining the free amino acids and organic acids in culture supernatants. It was found that the knockout mutant no longer synthesized 3-phenyllactic acid (PLA) and 4-hydroxyphenyllactic acid (HPLA). Additionally, the mutant strain no longer catabolized phenylalanine. Metabolic pathway analysis using the KEGG database indicate that P. acidilactici cannot synthesize α-ketoglutarate that is a predominant amino-group acceptor in many transamination reactions. To study the transfer of the amino group of phenylalanine, the wild-type strain was incubated with [15N] phenylalanine. Mass spectrometry showed that during fermentation, [15N] alanine was formed, indicating that pyruvic acid is an amino group acceptor in P. acidilactici. The present study shows that Aat plays a crucial role in PLA/HPLA biosynthesis and pyruvic acid is an amino acceptor in transamination reactions in P. acidilactici.
Item Type: |
Journal Article (Original Article) |
|---|---|
Division/Institute: |
School of Agricultural, Forest and Food Sciences HAFL School of Agricultural, Forest and Food Sciences HAFL > Consumer-focused Food Production > Bioconversion and Protective Cultures |
Name: |
Wenger, Alexander; Bär, Cornelia; Portmann, Reto; Schmidt, Remo S.; Eugster, Elisabeth  0000-0002-9183-5462; Weisskopf, Laure and Irmler, Stefan |
ISSN: |
1664-302X |
Publisher: |
Frontiers Research Foundation |
Language: |
English |
Submitter: |
Elisabeth Eugster |
Date Deposited: |
13 Jun 2023 15:46 |
Last Modified: |
20 Jun 2023 10:48 |
Publisher DOI: |
10.3389/fmicb.2023.1150425 |
ARBOR DOI: |
10.24451/arbor.19344 |
URI: |
https://arbor.bfh.ch/id/eprint/19344 |
Actions (login required)
 |
View Item |