Mechanism-Guided Design of Highly Efficient Protein Secretion and Lipid Conversion for Biomanufacturing and Biorefining

Shangxian Xie; Su Sun; Furong Lin; Muzi Li; Yunqiao Pu; Yanbing Cheng; Bing Xu; Zhihua Liu; Leonardo da Costa Sousa; Bruce E. Dale; Arthur J. Ragauskas; Susie Y. Dai; Joshua S. Yuan

doi:10.1002/advs.201801980

Mechanism-Guided Design of Highly Efficient Protein Secretion and Lipid Conversion for Biomanufacturing and Biorefining

Shangxian Xie, Su Sun, Furong Lin, Muzi Li, Yunqiao Pu, Yanbing Cheng, Bing Xu, Zhihua Liu, Leonardo da Costa Sousa, Bruce E. Dale, Arthur J. Ragauskas, Susie Y. Dai, Joshua S. Yuan

Biomaterials and Biomass Characterizatio

Research output: Contribution to journal › Article › peer-review

60 Scopus citations

Abstract

Bacterial protein secretion represents a significant challenge in biotechnology, which is essential for the cost-effective production of therapeutics, enzymes, and other functional proteins. Here, it is demonstrated that proteomics-guided engineering of transcription, translation, secretion, and folding of ligninolytic laccase balances the process, minimizes the toxicity, and enables efficient heterologous secretion with a total protein yield of 13.7 g L⁻¹. The secretory laccase complements the biochemical limits on lignin depolymerization well in Rhodococcus opacus PD630. Further proteomics analysis reveals the mechanisms for the oleaginous phenotype of R. opacus PD630, where a distinct multiunit fatty acid synthase I drives the carbon partition to storage lipid. The discovery guides the design of efficient lipid conversion from lignin and carbohydrate. The proteomics-guided integration of laccase-secretion and lipid production modules enables a high titer in converting lignin-enriched biorefinery waste to lipid. The fundamental mechanisms, engineering components, and design principle can empower transformative platforms for biomanufacturing and biorefining.

Original language	English
Article number	1801980
Journal	Advanced Science
Volume	6
Issue number	13
DOIs	https://doi.org/10.1002/advs.201801980
State	Published - Jul 3 2019

Funding

The authors thank Dr. Lindsay D. Eltis for the gift of plasmid pK18mobsacB and advising on the experiment of gene knockout in rhodococci. The work was supported by the U.S. DOE (Department of Energy) Office of Energy Efficiency and Renewable Energy (EERE) BETO (Bioenergy Technology Office) (Grant Nos. DE-EE0006112, DE-EE0007104, and DE-EE0008250) to J.S.Y. and A.R. The research was also supported by Texas A&M Agrilife Research's biofuel initiative to J.S.Y. The authors thank Dr. Lindsay D. Eltis for the gift of plasmid pK18mobsacB and advising on the experiment of gene knockout in rhodococci. The work was supported by the U.S. DOE (Department of Energy) Office of Energy Efficiency and Renewable Energy (EERE) BETO (Bioenergy Technology Office) (Grant Nos. DE-EE0006112, DE-EE0007104, and DE-EE0008250) to J.S.Y. and A.R. The research was also supported by Texas A&M Agrilife Research’s biofuel initiative to J.S.Y.

Funders	Funder number
Bioenergy Technology Office
U.S. DOE
National Science Foundation	1632976
U.S. Department of Energy
Texas AgriLife Research
Office of Energy Efficiency and Renewable Energy
Bioenergy Technologies Office	DE-EE0006112, DE-EE0007104, DE-EE0008250

Keywords

applied microbiology
heterologous protein secretion expression
lignin valorization
lipid biosynthesis
metabolic engineering

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10.1002/advs.201801980

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title = "Mechanism-Guided Design of Highly Efficient Protein Secretion and Lipid Conversion for Biomanufacturing and Biorefining",

abstract = "Bacterial protein secretion represents a significant challenge in biotechnology, which is essential for the cost-effective production of therapeutics, enzymes, and other functional proteins. Here, it is demonstrated that proteomics-guided engineering of transcription, translation, secretion, and folding of ligninolytic laccase balances the process, minimizes the toxicity, and enables efficient heterologous secretion with a total protein yield of 13.7 g L−1. The secretory laccase complements the biochemical limits on lignin depolymerization well in Rhodococcus opacus PD630. Further proteomics analysis reveals the mechanisms for the oleaginous phenotype of R. opacus PD630, where a distinct multiunit fatty acid synthase I drives the carbon partition to storage lipid. The discovery guides the design of efficient lipid conversion from lignin and carbohydrate. The proteomics-guided integration of laccase-secretion and lipid production modules enables a high titer in converting lignin-enriched biorefinery waste to lipid. The fundamental mechanisms, engineering components, and design principle can empower transformative platforms for biomanufacturing and biorefining.",

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author = "Shangxian Xie and Su Sun and Furong Lin and Muzi Li and Yunqiao Pu and Yanbing Cheng and Bing Xu and Zhihua Liu and {da Costa Sousa}, Leonardo and Dale, {Bruce E.} and Ragauskas, {Arthur J.} and Dai, {Susie Y.} and Yuan, {Joshua S.}",

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doi = "10.1002/advs.201801980",

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AU - Pu, Yunqiao

AU - Cheng, Yanbing

AU - Xu, Bing

AU - Liu, Zhihua

AU - da Costa Sousa, Leonardo

AU - Dale, Bruce E.

AU - Ragauskas, Arthur J.

AU - Dai, Susie Y.

AU - Yuan, Joshua S.

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AB - Bacterial protein secretion represents a significant challenge in biotechnology, which is essential for the cost-effective production of therapeutics, enzymes, and other functional proteins. Here, it is demonstrated that proteomics-guided engineering of transcription, translation, secretion, and folding of ligninolytic laccase balances the process, minimizes the toxicity, and enables efficient heterologous secretion with a total protein yield of 13.7 g L−1. The secretory laccase complements the biochemical limits on lignin depolymerization well in Rhodococcus opacus PD630. Further proteomics analysis reveals the mechanisms for the oleaginous phenotype of R. opacus PD630, where a distinct multiunit fatty acid synthase I drives the carbon partition to storage lipid. The discovery guides the design of efficient lipid conversion from lignin and carbohydrate. The proteomics-guided integration of laccase-secretion and lipid production modules enables a high titer in converting lignin-enriched biorefinery waste to lipid. The fundamental mechanisms, engineering components, and design principle can empower transformative platforms for biomanufacturing and biorefining.

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Mechanism-Guided Design of Highly Efficient Protein Secretion and Lipid Conversion for Biomanufacturing and Biorefining

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