Alkene cleavage activity of Pleurotus sapidus to obtain natural flavors

2020 ◽  
Vol 92 (9) ◽  
pp. 1338-1339
Author(s):  
N.-K. Krahe ◽  
A. B. Omarini ◽  
H. Zorn ◽  
R. G. Berger
Molecules ◽  
2020 ◽  
Vol 25 (7) ◽  
pp. 1536
Author(s):  
Nina-Katharina Krahe ◽  
Ralf G. Berger ◽  
Franziska Ersoy

Alkene cleavage is a possibility to generate aldehydes with olfactory properties for the fragrance and flavor industry. A dye-decolorizing peroxidase (DyP) of the basidiomycete Pleurotus sapidus (PsaPOX) cleaved the aryl alkene trans-anethole. The PsaPOX was semi-purified from the mycelium via FPLC, and the corresponding gene was identified. The amino acid sequence as well as the predicted tertiary structure showed typical characteristics of DyPs as well as a non-canonical Mn2+-oxidation site on its surface. The gene was expressed in Komagataella pfaffii GS115 yielding activities up to 142 U/L using 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) as substrate. PsaPOX exhibited optima at pH 3.5 and 40 °C and showed highest peroxidase activity in the presence of 100 µM H2O2 and 25 mM Mn2+. PsaPOX lacked the typical activity of DyPs towards anthraquinone dyes, but oxidized Mn2+ to Mn3+. In addition, bleaching of β-carotene and annatto was observed. Biotransformation experiments verified the alkene cleavage activity towards the aryl alkenes (E)-methyl isoeugenol, α-methylstyrene, and trans-anethole, which was increased almost twofold in the presence of Mn2+. The resultant aldehydes are olfactants used in the fragrance and flavor industry. PsaPOX is the first described DyP with alkene cleavage activity towards aryl alkenes and showed potential as biocatalyst for flavor production.


2021 ◽  
Vol 22 (3) ◽  
pp. 1363
Author(s):  
Nina-Katharina Krahe ◽  
Ralf G. Berger ◽  
Martin Witt ◽  
Holger Zorn ◽  
Alejandra B. Omarini ◽  
...  

The basidiomycete Pleurotus sapidus produced a dye-decolorizing peroxidase (PsaPOX) with alkene cleavage activity, implying potential as a biocatalyst for the fragrance and flavor industry. To increase the activity, a daughter-generation of 101 basidiospore-derived monokaryons (MK) was used. After a pre-selection according to the growth rate, the activity analysis revealed a stable intraspecific variability of the strains regarding peroxidase and alkene cleavage activity of PsaPOX. Ten monokaryons reached activities up to 2.6-fold higher than the dikaryon, with MK16 showing the highest activity. Analysis of the PsaPOX gene identified three different enzyme variants. These were co-responsible for the observed differences in activities between strains as verified by heterologous expression in Komagataella phaffii. The mutation S371H in enzyme variant PsaPOX_high caused an activity increase alongside a higher protein stability, while the eleven mutations in variant PsaPOX_low resulted in an activity decrease, which was partially based on a shift of the pH optimum from 3.5 to 3.0. Transcriptional analysis revealed the increased expression of PsaPOX in MK16 as reason for the higher PsaPOX activity in comparison to other strains producing the same PsaPOX variant. Thus, different expression profiles, as well as enzyme variants, were identified as crucial factors for the intraspecific variability of the PsaPOX activity in the monokaryons.


RSC Advances ◽  
2020 ◽  
Vol 10 (65) ◽  
pp. 39753-39762
Author(s):  
Yuichiro Otsuka ◽  
Masanobu Nojiri ◽  
Norihisa Kusumoto ◽  
Ronald R. Navarro ◽  
Koh Hashida ◽  
...  

This work explores the utilization of wood for high-value production of novel alcoholic brews and liquors with natural flavors.


2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Wenjing Qi ◽  
Erika D. V. Gromoff ◽  
Fan Xu ◽  
Qian Zhao ◽  
Wei Yang ◽  
...  

AbstractMulticellular organisms coordinate tissue specific responses to environmental information via both cell-autonomous and non-autonomous mechanisms. In addition to secreted ligands, recent reports implicated release of small RNAs in regulating gene expression across tissue boundaries. Here, we show that the conserved poly-U specific endoribonuclease ENDU-2 in C. elegans is secreted from the soma and taken-up by the germline to ensure germline immortality at elevated temperature. ENDU-2 binds to mature mRNAs and negatively regulates mRNA abundance both in the soma and the germline. While ENDU-2 promotes RNA decay in the soma directly via its endoribonuclease activity, ENDU-2 prevents misexpression of soma-specific genes in the germline and preserves germline immortality independent of its RNA-cleavage activity. In summary, our results suggest that the secreted RNase ENDU-2 regulates gene expression across tissue boundaries in response to temperature alterations and contributes to maintenance of stem cell immortality, probably via retaining a stem cell specific program of gene expression.


2021 ◽  
Vol 178 ◽  
pp. 113019
Author(s):  
Kai Zhang ◽  
Zhenqiang Fan ◽  
Bo Yao ◽  
Yuedi Ding ◽  
Jing Zhao ◽  
...  
Keyword(s):  

2021 ◽  
Vol 36 ◽  
pp. 127834
Author(s):  
Koichi Kato ◽  
Yoshimi Ichimaru ◽  
Yoshinori Okuno ◽  
Yoshihiro Yamaguchi ◽  
Wanchun Jin ◽  
...  

2021 ◽  
Author(s):  
Yiren Cao ◽  
Jinjun Wu ◽  
Bo Pang ◽  
Hongquan Zhang ◽  
X. Chris Le

The trans-cleavage activity of the target-activated CRISPR-Cas12a liberated an RNA crosslinker from a molecular transducer, which facilitated assembly of gold nanoparticles. Integration of the molecular transducer with isothermal amplification and...


Foods ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1545
Author(s):  
Valentina Maria Merlino ◽  
Daniela Fracassetti ◽  
Alessandra Di Canito ◽  
Simona Pizzi ◽  
Danielle Borra ◽  
...  

The consumption of traditional wine has decreased in Europe during the last fifteen years. In parallel, new wine alternatives obtained by blending wines and fruit juices or by flavoring wines with artificial or natural flavors have appeared on the market. Recently, an innovative fruit wine obtained by co-fermentation of grape must and kiwi juice has been proposed and its potential of attraction for consumers should be exploited. To assess the potential consumer acceptability and expectations towards this new product, an online choice experiment has been conducted involving a consumer group of young adults (18–35 years old; n = 373). After the data collection, participants were divided into two groups according to whether they had already tasted a fruit wine (neophiles) or had never tasted it (new entries). For each group, the individual’s responses (on wine consumption habits, expectations and willingness to consume and pay a fruit wine) were analyzed through Principal Component Analysis. Different consumption styles and expectation patterns were defined in the two groups. However, in general, neophiles showed consumption patterns based on the evaluation of fruit quality, sales format, alcoholic content and the presence or not of bubbles, not giving importance to the brand. In contrast, new entries’ responses identified consumption patterns driven by the willingness to pay for a new product, the product value for money and packaging features. Differences between the two groups in expectations about the product sensory characteristics also emerged. These findings should contribute to this area of study by integrating environmental, economic and social dimensions and addressing food innovation and sustainability in the fruit and wine chains.


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