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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.

Authors:
ORCiD logo [1];  [2];  [2];  [2]; ORCiD logo [3];  [2];  [2];  [2];  [4];  [4]; ORCiD logo [5];  [6]; ORCiD logo [2]
  1. Texas A & M Univ., College Station, TX (United States); Huazhong Univ. of Science and Technology, Wuhan (China
  2. Texas A & M Univ., College Station, TX (United States)
  3. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
  4. Michigan State Univ., East Lansing, MI (United States)
  5. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Univ. of Tennessee, Knoxville, TN (United States)
  6. Texas A & M Univ., College Station, TX (United States); Univ. of Iowa, Coralville, IA (United States). State Hygienic Lab.
Publication Date:
Research Org.:
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Sponsoring Org.:
USDOE Office of Energy Efficiency and Renewable Energy (EERE), Bioenergy Technologies Office (EE-3B)
OSTI Identifier:
1510339
Alternate Identifier(s):
OSTI ID: 1510340; OSTI ID: 1530062
Grant/Contract Number:  
AC05-00OR22725; EE0006112; EE0007104; EE0008250; DE‐EE0006112; DE‐EE0007104; DE‐EE0008250
Resource Type:
Published Article
Journal Name:
Advanced Science
Additional Journal Information:
Journal Volume: TBD; Journal Issue: TBD; Journal ID: ISSN 2198-3844
Publisher:
Wiley
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; applied microbiology; heterologous protein secretion expression; lignin valorization; lipid biosynthesis; metabolic engineering

Citation Formats

Xie, Shangxian, Sun, Su, Lin, Furong, Li, Muzi, Pu, Yunqiao, Cheng, Yanbing, Xu, Bing, Liu, Zhihua, da Costa Sousa, Leonardo, Dale, Bruce E., Ragauskas, Arthur J., Dai, Susie Y., and Yuan, Joshua S. Mechanism-Guided Design of Highly Efficient Protein Secretion and Lipid Conversion for Biomanufacturing and Biorefining. United States: N. p., 2019. Web. doi:10.1002/advs.201801980.
Xie, Shangxian, Sun, Su, Lin, Furong, Li, Muzi, Pu, Yunqiao, Cheng, Yanbing, Xu, Bing, Liu, Zhihua, da Costa Sousa, Leonardo, Dale, Bruce E., Ragauskas, Arthur J., Dai, Susie Y., & Yuan, Joshua S. Mechanism-Guided Design of Highly Efficient Protein Secretion and Lipid Conversion for Biomanufacturing and Biorefining. United States. doi:10.1002/advs.201801980.
Xie, Shangxian, Sun, Su, Lin, Furong, Li, Muzi, Pu, Yunqiao, Cheng, Yanbing, Xu, Bing, Liu, Zhihua, da Costa Sousa, Leonardo, Dale, Bruce E., Ragauskas, Arthur J., Dai, Susie Y., and Yuan, Joshua S. Wed . "Mechanism-Guided Design of Highly Efficient Protein Secretion and Lipid Conversion for Biomanufacturing and Biorefining". United States. doi:10.1002/advs.201801980.
@article{osti_1510339,
title = {Mechanism-Guided Design of Highly Efficient Protein Secretion and Lipid Conversion for Biomanufacturing and Biorefining},
author = {Xie, Shangxian and Sun, Su and Lin, Furong and Li, Muzi and Pu, Yunqiao and Cheng, Yanbing and Xu, Bing and Liu, Zhihua and da Costa Sousa, Leonardo and Dale, Bruce E. and Ragauskas, Arthur J. and Dai, Susie Y. and Yuan, Joshua S.},
abstractNote = {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.},
doi = {10.1002/advs.201801980},
journal = {Advanced Science},
number = TBD,
volume = TBD,
place = {United States},
year = {2019},
month = {5}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
DOI: 10.1002/advs.201801980

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Works referenced in this record:

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