Engineering cofactor supply and recycling to drive phenolic acid biosynthesis in yeast

Chen, Ruibing; Gao, Jiaoqi; Wu, Yu; Chen, Xianghui; Zhai, Xiaoxin; Yu, Chen; Zhang, Lei; Zhou, Yongjin J.

doi:10.1038/s41589-022-01014-6

Cited by 97 publications

(129 citation statements)

References 61 publications

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“…In 2022, on the basis of pathway optimization in yeast S. cerevisiae , Chen et al manipulated biosynthesis, compartmentalization, and recycling of two cofactors, NADPH and S -adenosyl- l -methion (SAM), which participate in redox reactions and methylation reactions, respectively. The production of the phenylpropane pathway intermediates CaA (5.5 ± 0.2 g L −1 ) and FA (3.8 ± 0.3 g L −1 ) reached gram-level production in the fed-batch fermentation, which has a considerable production potential at present 33 (Fig. 2).…”

Section: Biosynthesismentioning

confidence: 94%

“…31,32 The upstream compounds caffeic acid (CaA) and ferulic acid (FA) were achieved in gram-level production (5.5 AE 0.2 g L À1 CaA, 3.8 AE 0.3 g L À1 FA) in microbial cell factories by engineering cofactor supply and recycling. 33 Moreover, the catalytic mechanism of pathway enzymes, such as dirigent proteins (DIRs), secoisolariciresinol dehydrogenase (SDH), and deoxypodophyllotoxin synthase (DPS) have been revealed, enhancing understanding of a growing number of biochemical transformations.…”

Section: Introductionmentioning

confidence: 99%

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Biosynthesis, total synthesis, and pharmacological activities of aryltetralin-type lignan podophyllotoxin and its derivatives

et al. 2022

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Section: Biosynthesismentioning

confidence: 94%

Section: Introductionmentioning

confidence: 99%

Biosynthesis, total synthesis, and pharmacological activities of aryltetralin-type lignan podophyllotoxin and its derivatives

et al. 2022

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“…Clade-2 also include commonly used Streptomyces chassis, golden standard Streptomyces coelicolor [29] and fast growing Streptomyces venezuelae [32]. Also, 5 out of the 8 strains examined in this study, including A41733, ATCC23862, ATCC31561, ATCC31713 and A44034, belong to clade-2 [43][44][45], hinting at the potential of these strains as alternative chassis for NP production.…”

Section: Resultsmentioning

confidence: 99%

Cost-effective hybrid long-short read assembly delineates alternative GC-richStreptomyceschassis for natural product discovery

Heng¹,

Tan²,

Tay

et al. 2022

Preprint

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With the advent of rapid automated in silico identification of biosynthetic gene clusters (BGCs), genomics presents vast opportunities to accelerate natural product (NP) discovery. However, prolific NP producers, Streptomyces, are exceptionally GC-rich (>80%) and highly repetitive within BGCs. These pose challenges in sequencing and high-quality genome assembly which are currently circumvented via intensive sequencing. Here, we outline a more cost-effective workflow using multiplex Illumina and Oxford Nanopore sequencing with hybrid long-short read assembly algorithms to generate high quality genomes. Our protocol involves subjecting long read derived assemblies to up to 4 rounds of polishing with short reads to yield accurate BGC predictions. We successfully sequenced and assembled 8 GC-rich Streptomyces genomes whose lengths range from 7.1 to 12.1 Mb at an average N50 of 5.9 Mb. Taxonomic analysis revealed previous misrepresentation among these strains and allowed us to propose a potentially new species, Streptomyces sydneybrenneri. Further comprehensive characterization of their biosynthetic, pan-genomic and antibiotic resistance features especially for molecules derived from type I polyketide synthase (PKS) BGCs reflected their potential as NP chassis. Thus, the genome assemblies and insights presented here are envisioned to serve as gateway for the scientific community to expand their avenues in NP discovery.

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“…However, most studies focused on the discovery and mining of novel P450s 37 , the improvement of P450 catalytic properties 38 and the enhancement of expression levels in microbial hosts 39 . For most of enzymes, the su cient supply of essential cofactors is important for their catalytic e ciencies 40 . Especially for P450s, the apoenzymes will completely lose their activities and are prone to aggregation or degradation when the level of intracellular heme is insu cient 17 .…”

Section: Discussionmentioning

confidence: 99%

Whole-cell P450 biocatalysts using engineered Escherichia coli with fine-tuned heme supply

Zhou

et al. 2022

Preprint

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By exploiting versatile P450 enzymes, whole-cell biocatalysis can be performed to synthesize valuable compounds in Escherichia coli. However, the insufficient supply of heme limits the whole-cell P450 biocatalytic activity. Here we report a strategy of improving intracellular heme supply for enhancing the catalytic efficiencies of P450s. After comparing the effects of enhancing heme transport and biosynthesis on P450 activities, intracellular heme supply was optimized through the integrated expression of necessary synthetic genes at proper ratio and the assembly of rate-limiting enzymes using DNA-guided scaffolds. By the combined use of mutated heme-sensitive biosensor and small regulatory RNA systems, the intracellular heme level was fine-tuned. The catalytic efficiencies of three different P450s, BM3, sca-2 and CYP105D7, were enhanced through fine-tuning heme supply for the synthesis of hydroquinone, pravastatin, and 7,3',4'-trihydroxyisoflavone as examples of chemical intermediate, drug, and natural product, respectively. This strategy will be useful to produce other hemoproteins with high activities.

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Engineering cofactor supply and recycling to drive phenolic acid biosynthesis in yeast

Cited by 97 publications

References 61 publications

Biosynthesis, total synthesis, and pharmacological activities of aryltetralin-type lignan podophyllotoxin and its derivatives

Biosynthesis, total synthesis, and pharmacological activities of aryltetralin-type lignan podophyllotoxin and its derivatives

Cost-effective hybrid long-short read assembly delineates alternative GC-richStreptomyceschassis for natural product discovery

Whole-cell P450 biocatalysts using engineered Escherichia coli with fine-tuned heme supply

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