| RRC ID |
84651
|
| Author |
Kasahara E, Kitamura Y, Katada M, Mizuki M, Okumura N, Sano T, Koizumi Y, Maeda K, Takahashi-Ando N, Kimura M, Nakajima Y.
|
| Title |
Attempting to Create a Pathway to 15-Deacetylcalonectrin with Limited Accumulation in Cultures of Fusarium Tri3 Mutants: Insight into Trichothecene Biosynthesis Machinery.
|
| Journal |
Int J Mol Sci
|
| Abstract |
The compound 15-deacetylcalonectrin (15-deCAL) is a common pathway intermediate in the biosynthesis of Fusarium trichothecenes. This tricyclic intermediate is metabolized to calonectrin (CAL) by trichothecene 15-O-acetyltransferase encoded by Tri3. Unlike other trichothecene pathway Tri gene mutants, the Δtri3 mutant produces lower amounts of the knocked-out enzyme's substrate 15-deCAL, and instead, accumulates higher quantities of earlier bicyclic intermediate and shunt metabolites. Furthermore, evolutionary studies suggest that Tri3 may play a role in shaping the chemotypes of trichothecene-producing Fusarium strains. To better understand the functional role of Tri3p in biosynthesis and evolution, we aimed to develop a method to produce 15-deCAL by using transgenic Fusarium graminearum strains derived from a trichothecene overproducer. Unfortunately, introducing mutant Tri3, encoding a catalytically impaired but structurally intact acetylase, did not improve the low 15-deCAL production level of the ΔFgtri3 deletion strain, and the bicyclic products continued to accumulate as the major metabolites of the active-site mutant. These findings are discussed in light of the enzyme responsible for 15-deCAL production in trichothecene biosynthesis machinery. To efficiently produce 15-deCAL, we tested an alternative strategy of using a CAL-overproducing transformant. By feeding a crude CAL extract to a Fusarium commune strain that was isolated in this study and capable of specifically deacetylating C-15 acetyl, 15-deCAL was efficiently recovered. The substrate produced in this manner can be used for kinetic investigations of this enzyme and its possible role in chemotype diversification.
|
| Volume |
25(12)
|
| Published |
2024-6-11
|
| DOI |
10.3390/ijms25126414
|
| PII |
ijms25126414
|
| PMID |
38928120
|
| PMC |
PMC11203908
|
| MeSH |
Acetyltransferases / genetics
Acetyltransferases / metabolism
Biosynthetic Pathways / genetics
Fungal Proteins / genetics
Fungal Proteins / metabolism
Fusarium* / genetics
Fusarium* / metabolism
Mutation*
Trichothecenes* / metabolism
|
| Resource |
| General Microbes |
JCM9873 |
