Engineering Mycolicibacterium neoaurum for the production of antioxidant ergothioneine

Abstract

Ergothioneine (EGT) represents valuable protective functions for humans, but EGT from the diet cannot meet daily requirements. Although the heterologous synthesis of EGT had been achieved, it is still a challenge to obtain stable and highyield EGT-producing cell factories. Here, after the co-overexpression of the EGT synthetic gene cluster and hisG, hisC, and allB1 in Mycolicibacterium neoaurum, the natural EGT titer was increased by 7.2-folds. However, the degradation problem of EGT in large-scale fermentation needs to be urgently solved. A putative lyase gene Mn_3042 was inactivated, thus inhibiting the product degradation and increasing the EGT titer by 21%. Moreover, the enhancement of S-adenosyl-L-methionine regeneration further increased EGT titer by 28%. After optimization of fed-batch fermentation, the yield of EGT was boosted to 1.56 g/L with a productivity of 7.2 mg/L/h. This study provides a systematic engineering strategy for developing EGT-producing cell factories.

Engineering Methyltransferase and Sulfoxide Synthase for High-Yield Production of Ergothioneine

“…In addition to E. coli, many other organisms have also been engineered for heterologous production of ERG, such as Saccharomyces cerevisiae, Corynebacterium glutamicum, Mycolicibacterium neoaurum, and so on. − van der Hoek et al engineered the amino acid metabolism of S. cerevisiae and expressed the combinatorial ERG synthesis pathway in the engineered strain.…”

Co‐augmentation of a transport gene mfsT1 in Mycolicibacterium neoaurum with genome engineering to enhance ergothioneine production

“…Media and culture conditions in the previous study [22] were used here. E. coli grew in Luria-Bertani (LB) medium at 37°C.…”

“…Hence, to mine potential transporters to increase EGT transmembrane transport in M. neoaurum, the comparative transcriptome data were re-analyzed between the EGT10B and the wild-type strain. In brief, compared with the wild-type strain, a total of 215 genes showed significant upregulation in EGT10B (Log2 fold change ratio ≥1.0, the gene expression upregulation ≥2.0-folds), whereas 108 genes showed significant downregulation (Log2 fold change ratio ≤−1.0, the gene expression downregulation ≥2.0-folds) (Figure 2a) [22]. Among the above genes, we focused on the major facilitator superfamily (MFS), the largest class of secondary active transporters.…”

“…In our previous study [22], we engineered Mycolicibacterium neoaurum into a high‐capacity cell factory for EGT production. The resulting strain achieved an EGT yield of 148.2 ± 13.7 mg/L in 250‐mL shake flask cultivation.…”

ERGOT Claviceps purpurea (Fries) Tulasne ALKALOID DIVERSITY AND VIRULENCE: EVOLUTION, GENETIC DIVERSIFICATION, AND METABOLIC ENGINEERING (review)

“…Таким образом, на сорвеменном этапе исследований C. purpureaэто, с одной стороны, важнейший продуцент большого количества биологически активных соединений (алкалоидов) и уникальная модель системы паразит-хозяин (218)(219)(220)(221), с другой -патоген, наносящий значительный экономический ущерб производителям зерна, кормовой и животноводческой отрасли по всему миру (222,223). В то время как медицинские и биотехнологические исследования сосредоточены на положительных эффектах алкалоидов (224) и других вторичных метаболитов спорыньи и особенностях их получения (224)(225)(226)(227)(228), в растениеводстве, животноводстве и пищевой промышленности опасность эргоалкалоидов остается острой и вызывающей беспокойство общемировой проблемой (229)(230)(231)(232). Ее серьезность усугубляется тем, что в меняющихся условиях окружающей среды у Claviceps изменяются профили и интенсивность продукции алкалоидов (231).…”