Tryptophol (TOL) is a metabolic derivative of tryptophan
(Trp) and shows pleiotropic effects in humans, plants and
microbes. In this study, the effect of Trp and phenylalanine
(Phe) on TOL production in Saccharomyces cerevisiae was determined,
and a systematic interpretation of TOL accumulation
was offered by transcriptomic and metabolomic analyses.
Trp significantly promoted TOL production, but the output
plateaued (231.02−266.31 mg/L) at Trp concentrations ≥ 0.6
g/L. In contrast, Phe reduced the stimulatory effect of Trp,
which was strongly dependent on the Phe concentration. An
integrated genomic, transcriptomic, and metabolomic analysis
revealed that the effect of Trp and Phe on TOL production
was mainly related to the transamination and decarboxylation
of the Ehrlich pathway. Additionally, other genes, including
thiamine regulon genes (this), the allantoin catabolic
genes dal1, dal2, dal4, and the transcriptional activator gene
aro80, may play important roles. These findings were partly
supported by the fact that the thi4 gene was involved in TOL
production, as shown by heterologous expression analysis. To
the best of our knowledge, this novel biological function of thi4
in S. cerevisiae is reported here for the first time. Overall, our
findings provide insights into the mechanism of TOL production,
which will contribute to TOL production using metabolic
engineering strategies.
Arthrobotrys oligospora is a model species of nematophagous
fungi and has great potential for the biological control of nematode
diseases. Lectin is a protein that binds to carbohydrates
and their complexes with high specificity, which mediates recognition
events in various physiological and pathological
processes. This study aimed to investigate the role of the
Jacalin-related lectin (JRL) gene, AOL_s00083g511, in A. oligospora
development. Through a homology recombination
approach, we obtained the AOL_s00083g511 knockout mutant
strain (Δg511). Next, the biological characteristics of the
Δg511 mutant strain, including growth rate, conidia germination
rate, adaptation to environmental stresses, and nematocidal
activity, were compared with those of the wild-type
(WT) strain. The results showed that the JRL gene AOL_
s00083g511 did not affect fungal growth, conidia germination,
3D-trap formation, and the ability of A. oligospora to
prey on nematodes significantly. We speculate that this phenomenon
may be caused by a loss of the key β1–β2 loops in
the AOL_ s00083g511-encoded JRL domain and an intrinsic
genetic compensation of AOL_s00083g511 in this fungus.
The growth rates of both strains on high salt or surfactant media
were similar; however, in the strong oxidation medium,
the growth rate of the Δg511 mutant was significantly lower
than that of the WT strain, indicating that AOL_s00083g511
might play a role in oxidative stress resistance. These findings
provide a basis for further analysis of the related functions
of the JRL gene in A. oligospora and their potential roles
in the biological control of nematodes in the future.
Newcastle disease, designated a class A disease of poultry by
the Office international des epizooties (OIE), is an acute infection
caused by Newcastle disease virus (NDV). The merging
of the envelope of NDV with the membrane of a target
host cell is the key step in the infection pathway, which is driven
by the concerted action of two glycoproteins: haemagglutinin-
neuraminidase (HN) and fusion (F) protein. When
the HN protein binds to the host cell surface receptor, the F
protein is activated to mediate fusion. The three-dimensional
structure of the F protein has been reported to have low
electron density between the DIII domain and the HRB domain,
and this electron-poor region is defined as the HRB
linker. To clarify the contributing role of the HRB linker in
the NDV F protein-mediated fusion process, 6 single amino
acid mutants were obtained by site-directed mutagenesis of
the HRB linker. The expression of the mutants and their abilities
to mediate fusion were analysed, and the key amino acids
in the HRB linker were identified as L436, E439, I450, and
S453, as they can modulate the fusion ability or expression
of the active form to a certain extent. The data shed light on
the crucial role of the F protein HRB linker in the acquisition
of a normal fusogenic phenotype.
The Gram-positive bacterium Enterococcus faecalis is currently
one of the major pathogens of nosocomial infections.
The lifestyle of E. faecalis relies primarily on its remarkable capacity
to face and survive in harsh environmental conditions.
Toxin-antitoxin (TA) systems have been linked to the growth
control of bacteria in response to adverse environments but
have rarely been reported in Enterococcus. Three functional
type II TA systems were identified among the 10 putative
TA systems encoded by E. faecalis ATCC29212. These toxin
genes have conserved domains homologous to MazF (DR75_
1948) and ImmA/IrrE family metallo-endopeptidases (DR75_
1673 and DR75_2160). Overexpression of toxin genes could
inhibit the growth of Escherichia coli. However, the toxin
DR75_1673 could not inhibit bacterial growth, and the bacteriostatic
effect occurred only when it was coexpressed with
the antitoxin DR75_1672. DR75_1948–DR75_1949 and DR75_
160–DR75_2161 could maintain the stable inheritance of the
unstable plasmid pLMO12102 in E. coli. Moreover, the transcription
levels of these TAs showed significant differences
when cultivated under normal conditions and with different
temperatures, antibiotics, anaerobic agents and H2O2. When
DR75_2161 was knocked out, the growth of the mutant strain
at high temperature and oxidative stress was limited. The experimental
characterization of these TAs loci might be helpful
to investigate the key roles of type II TA systems in the
physiology and environmental stress responses of Enterococcus.