Microbial chassis engineering drives heterologous production of complex secondary metabolites

Liu, Jiaqi; Wang, Xue; Dai, Guangzhi; Zhang, Youming; Bian, Xiaoying

doi:10.1016/j.biotechadv.2022.107966

Cited by 36 publications

(30 citation statements)

References 261 publications

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“…Secondary metabolites are biologically active compounds synthesized by microorganisms using primary metabolites as precursors, which may be involved in the physiological processes of the bacterium itself and may also provide defense and resistance to external stressful environments ( Liu et al, 2022 ). The most common secondary metabolic gene clusters are type I, II, and III polyketides synthase (PKS) and non-ribosomal peptides synthase (NRPS) ( Medema et al, 2011 ).…”

Section: Resultsmentioning

confidence: 99%

Complete genome sequence, metabolic model construction, and huangjiu application of Saccharopolyspora rosea A22, a thermophilic, high amylase and glucoamylase actinomycetes

Liu²,

Han³

et al. 2022

Front. Microbiol.

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Saccharopolyspora is an important microorganism in the fermentation process of wheat qu and huangjiu, yet the mechanisms by which it performs specific functions in huangjiu remain unclear. A strain with high amylase and glucoamylase activities was isolated from wheat qu and identified as Saccharopolyspora rosea (S. rosea) A22. We initially reported the whole genome sequence of S. rosea A22, which comprised a circular chromosome 6,562,638 bp in size with a GC content of 71.71%, and 6,118 protein-coding genes. A functional genomic analysis highlighted regulatory genes involved in adaptive mechanisms to harsh conditions, and in vitro experiments revealed that the growth of S. rosea A22 could be regulated in response to the stress condition. Based on whole-genome sequencing, the first genome-scale metabolic model of S. rosea A22 named iSR1310 was constructed to predict the growth ability on different media with 91% accuracy. Finally, S. rosea A22 was applied to huangjiu fermentation by inoculating raw wheat qu, and the results showed that the total higher alcohol content was reduced by 12.64% compared with the control group. This study has elucidated the tolerance mechanisms and enzyme-producing properties of S. rosea A22 at the genetic level, providing new insights into its application to huangjiu.

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Section: Resultsmentioning

confidence: 99%

Complete genome sequence, metabolic model construction, and huangjiu application of Saccharopolyspora rosea A22, a thermophilic, high amylase and glucoamylase actinomycetes

Liu²,

Han³

et al. 2022

Front. Microbiol.

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“…Abundant secondary metabolites indicated that it could provide abundant substrates for expressing heterologous BGCs. In addition to possessing huge biosynthetic potential, strain E264 also has a fast growth rate, which makes it more suitable as a heterologous host ( Mao et al, 2017 ; Liu et al, 2022 ). However, as a member of the genus Burkholderia , the intrinsic multidrug-resistance of the wild-type strain E264 makes genetic manipulation difficult, which partially results from the resistance-nodulation-division family efflux pumps ( Biot et al, 2011 , 2013 ).…”

Section: Introductionmentioning

confidence: 99%

Engineering of Burkholderia thailandensis strain E264 serves as a chassis for expression of complex specialized metabolites

et al. 2022

Self Cite

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Heterologous expression is an indispensable approach to exploiting natural products from phylogenetically diverse microbial communities. In this study, we constructed a heterologous expression system based on strain Burkholderia thailandensis E264 by deleting efflux pump genes and screening constitutive strong promoters. The biosynthetic gene cluster (BGC) of disorazol from Sorangium cellulosum So ce12 was expressed successfully with this host, and the yield of its product, disorazol F2, rather than A1, was improved to 38.3 mg/L by promoter substitution and insertion. In addition to the disorazol gene cluster, the BGC of rhizoxin from Burkholderia rhizoxinica was also expressed efficiently, whereas no specific peak was detected when shuangdaolide BGC from Streptomyces sp. B59 was transformed into the host. This system provides another option to explore natural products from different phylogenetic taxa.

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“…During this assembly process, each NRPS has to interact with its upstream and/or downstream NRPS(s) to transfer the monomer substrate or the growing peptidyl chain intermediates. Previous studies have reported the use of heterologous hosts (e.g., Escherichia coli , yeast, and Streptomyces ) to produce NRPs by recombinant coexpression of the related NRPS enzymes (Li & Neubauer, 2014; J. Liu, Wang, et al, 2022; Tippelt & Nett, 2021; Zhang et al, 2022). However, these product yields are often low even after a series of systematic optimizations, for instance, strain engineering, precursor feeding, and high cell density cultivation.…”

Section: Introductionmentioning

confidence: 99%

Stapled NRPS enhances the production of valinomycin in Escherichia coli

Huang

Liu

et al. 2022

Biotech & Bioengineering

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Nonribosomal peptides (NRPs) are a large family of secondary metabolites with notable bioactivities, which distribute widely in natural resources across microbes and plants. To obtain these molecules, heterologous production of NRPs in robust surrogate hosts like Escherichia coli represent a feasible approach. However, reconstitution of the full biosynthetic pathway in a host often leads to low productivity, which is at least in part due to the low efficiency of enzyme interaction in vivo except for the well‐known reasons of metabolic burden (e.g., expression of large NRP synthetases—NRPSs with molecular weights of >100 kDa) and cellular toxicity on host cells. To enhance the catalytic efficiency of large NRPSs in vivo, here we propose to staple NRPS enzymes by using short peptide/protein pairs (e.g., SpyTag/SpyCatcher) for enhanced NRP production. We achieve this goal by introducing a stapled NRPS system for the biosynthesis of the antibiotic NRP valinomycin in E. coli. The results indicate that stapled valinomycin synthetase (Vlm1 and Vlm2) enables higher product accumulation than those two free enzymes (e.g., the maximum improvement is nearly fourfold). After further optimization by strain and bioprocess engineering, the final valinomycin titer maximally reaches about 2800 µg/L, which is 73 times higher than the initial titer of 38 µg/L. We expect that stapling NRPS enzymes will be a promising catalytic strategy for high‐level biosynthesis of NRP natural products.

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Microbial chassis engineering drives heterologous production of complex secondary metabolites

Cited by 36 publications

References 261 publications

Complete genome sequence, metabolic model construction, and huangjiu application of Saccharopolyspora rosea A22, a thermophilic, high amylase and glucoamylase actinomycetes

Complete genome sequence, metabolic model construction, and huangjiu application of Saccharopolyspora rosea A22, a thermophilic, high amylase and glucoamylase actinomycetes

Engineering of Burkholderia thailandensis strain E264 serves as a chassis for expression of complex specialized metabolites

Stapled NRPS enhances the production of valinomycin in Escherichia coli

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