Data for Cloning and Characterization of a Panel of Mitochondrial Targeting Sequences for Compartmentalization Engineering in Saccharomyces cerevisiae

Dong, Chang; Shi, Zhuwei; Huang, Lei; Zhao, Huimin; Xu, Zhinan; Lian, Jiazhang

doi:10.13012/B2IDB-3316538_V1

Title: Data for Cloning and Characterization of a Panel of Mitochondrial Targeting Sequences for Compartmentalization Engineering in Saccharomyces cerevisiae

Dataset
Other Related Research

Abstract

Mitochondrion is generally considered as the most promising subcellular organelle for compartmentalization engineering. Much progress has been made in reconstituting whole metabolic pathways in the mitochondria of yeast to harness the precursor pools (i.e., pyruvate and acetyl-CoA), bypass competing pathways, and minimize transportation limitations. However, only a few mitochondrial targeting sequences (MTSs) have been characterized (i.e., MTS of COX4), limiting the application of compartmentalization engineering for multigene biosynthetic pathways in the mitochondria of yeast. In the present study, based on the mitochondrial proteome, a total of 20 MTSs were cloned and the efficiency of these MTSs in targeting heterologous proteins, including the Escherichia coli FabI and enhanced green fluorescence protein (EGFP) into the mitochondria was evaluated by growth complementation and confocal microscopy. After systematic characterization, six of the well-performed MTSs were chosen for the colocalization of complete biosynthetic pathways into the mitochondria. As proof of concept, the full α-santalene biosynthetic pathway consisting of 10 expression cassettes capable of converting acetyl-coA to α-santalene was compartmentalized into the mitochondria, leading to a 3.7-fold improvement in the production of α-santalene. The newly characterized MTSs should contribute to the expanded metabolic engineering and synthetic biology toolbox for yeast mitochondrial compartmentalization engineering.

Authors:

Dong, Chang; Shi, Zhuwei; Huang, Lei; Zhao, Huimin; Xu, Zhinan; Lian, Jiazhang

Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering ,Zhejiang University, Hangzhou, China; Hangzhou Global Scientific and Technological Innovation Center, Zhejiang University, Hangzhou, China
Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering ,Zhejiang University, Hangzhou, China
Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana‐Champaign, Urbana, Illinois, USA; Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)
Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering ,Zhejiang University, Hangzhou, China; Hangzhou Global Scientific and Technological Innovation Center, Zhejiang University, Hangzhou, China; Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana‐Champaign, Urbana, Illinois, USA; Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)

Publication Date:: Fri Jul 16 20:00:00 EDT 2021

DOE Contract Number:: SC0018420

Research Org.:: Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States); University of Illinois Urbana-Champaign

Sponsoring Org.:: U.S. Department of Energy (DOE)

Subject:: Conversion; Metabolic Engineering

OSTI Identifier:: 3014140

DOI:: https://doi.org/10.13012/B2IDB-3316538_V1

Citation Formats


                    Dong, Chang, Shi, Zhuwei, Huang, Lei, Zhao, Huimin, Xu, Zhinan, and Lian, Jiazhang. Data for Cloning and Characterization of a Panel of Mitochondrial Targeting Sequences for Compartmentalization Engineering in Saccharomyces cerevisiae.  United States: N. p., 2021. 
        Web.  doi:10.13012/B2IDB-3316538_V1.

Copy to clipboard


                    Dong, Chang, Shi, Zhuwei, Huang, Lei, Zhao, Huimin, Xu, Zhinan, & Lian, Jiazhang. Data for Cloning and Characterization of a Panel of Mitochondrial Targeting Sequences for Compartmentalization Engineering in Saccharomyces cerevisiae.  United States.  doi:https://doi.org/10.13012/B2IDB-3316538_V1

Copy to clipboard


                    Dong, Chang, Shi, Zhuwei, Huang, Lei, Zhao, Huimin, Xu, Zhinan, and Lian, Jiazhang. 2021.  
"Data for Cloning and Characterization of a Panel of Mitochondrial Targeting Sequences for Compartmentalization Engineering in Saccharomyces cerevisiae".  United States.  doi:https://doi.org/10.13012/B2IDB-3316538_V1.  https://www.osti.gov/servlets/purl/3014140. Pub date:Fri Jul 16 20:00:00 EDT 2021

Copy to clipboard


                    
@article{osti_3014140,

  title        = {Data for Cloning and Characterization of a Panel of Mitochondrial Targeting Sequences for Compartmentalization Engineering in Saccharomyces cerevisiae},

  author       = {Dong, Chang and Shi, Zhuwei and Huang, Lei and Zhao, Huimin and Xu, Zhinan and Lian, Jiazhang},

  abstractNote = {Mitochondrion is generally considered as the most promising subcellular organelle for compartmentalization engineering. Much progress has been made in reconstituting whole metabolic pathways in the mitochondria of yeast to harness the precursor pools (i.e., pyruvate and acetyl-CoA), bypass competing pathways, and minimize transportation limitations. However, only a few mitochondrial targeting sequences (MTSs) have been characterized (i.e., MTS of COX4), limiting the application of compartmentalization engineering for multigene biosynthetic pathways in the mitochondria of yeast. In the present study, based on the mitochondrial proteome, a total of 20 MTSs were cloned and the efficiency of these MTSs in targeting heterologous proteins, including the Escherichia coli FabI and enhanced green fluorescence protein (EGFP) into the mitochondria was evaluated by growth complementation and confocal microscopy. After systematic characterization, six of the well-performed MTSs were chosen for the colocalization of complete biosynthetic pathways into the mitochondria. As proof of concept, the full α-santalene biosynthetic pathway consisting of 10 expression cassettes capable of converting acetyl-coA to α-santalene was compartmentalized into the mitochondria, leading to a 3.7-fold improvement in the production of α-santalene. The newly characterized MTSs should contribute to the expanded metabolic engineering and synthetic biology toolbox for yeast mitochondrial compartmentalization engineering.},

  doi          = {10.13012/B2IDB-3316538_V1},

  journal      = {},

  number       = ,

  volume       = ,

  place        = {United States},

  year         = {Fri Jul 16 20:00:00 EDT 2021},

  month        = {Fri Jul 16 20:00:00 EDT 2021}

}

Copy to clipboard

Dataset:

View Dataset

DOI: https://doi.org/10.13012/B2IDB-3316538_V1

Save / Share:

Export Metadata

Save to My Library

Similar records in DOE Data Explorer and OSTI.GOV collections:

Cloning and characterization of a panel of mitochondrial targeting sequences for compartmentalization engineering in Saccharomyces cerevisiae

Journal Article Dong, Chang ; Shi, Zhuwei ; Huang, Lei ; ... - Biotechnology and Bioengineering

Mitochondrion is generally considered as the most promising subcellular organelle for compartmentalization engineering. Much progress has been made in reconstituting whole metabolic pathways in the mitochondria of yeast to harness the precursor pools (i.e., pyruvate and acetyl-CoA), bypass competing pathways, and minimize transportation limitations. However, only a few mitochondrial targeting sequences (MTSs) have been characterized (i.e., MTS of COX4), limiting the application of compartmentalization engineering for multigene biosynthetic pathways in the mitochondria of yeast. In the present study, based on the mitochondrial proteome, a total of 20 MTSs were cloned and the efficiency of these MTSs in targeting heterologous proteins,more »« less
Data for L-Malic Acid Production from Xylose by Engineered Saccharomyces cerevisiae

Dataset Kang, Nam Kyu ; Lee, Jaewon ; Ort, Donald ; ...

L-malic acid is widely used in the food, chemical, and pharmaceutical industries. Here, we report on production of malic acid from xylose, the second most abundant sugar in lignocellulosic hydrolysates, by engineered Saccharomyces cerevisiae. To enable malic acid production in a xylose-assimilating S. cerevisiae, we overexpressed PYC1 and PYC2, coding for pyruvate carboxylases, a truncated MDH3 coding for malate dehydrogenase, and SpMAE1, coding for a Schizosaccharomyces pombe malate transporter. Additionally, both the ethanol- and glycerol-producing pathways were blocked to enhance malic acid production. The resulting strain produced malic acid from both glucose and xylose, but it produced much higher titersmore »« less
Data for Controlling Circuitry Underlies the Growth Optimization of Saccharomyces cerevisiae

Dataset Nguyen, Viviana ; Xue, Pu ; Li, Yifei ; ...

Microbial growth emerges from coordinated synthesis of various cellular components from limited resources. In Saccharomyces cerevisiae, cyclic AMP (cAMP)-mediated signaling is shown to orchestrate cellular metabolism; however, it remains unclear quantitatively how the controlling circuit drives resource partition and subsequently shapes biomass growth. Here we combined experiment with mathematical modeling to dissect the signaling-mediated growth optimization of S. cerevisiae. We showed that, through cAMP-mediated control, the organism achieves maximal or nearly maximal steady-state growth during the utilization of multiple tested substrates as well as under perturbations impairing glucose uptake. However, the optimal cAMP concentration varies across cases, suggesting that differentmore »« less
Data from Enhanced 2′-Fucosyllactose Production by Engineered Saccharomyces cerevisiae using Xylose as a Co-Substrate

Dataset Lee, Jaewon ; Kwak, Suryang ; Liu, Jing-Jing ; ...

2′-Fucosyllactose (2′-FL), a human milk oligosaccharide with confirmed benefits for infant health, is a promising infant formula ingredient. Although Escherichia coli, Saccharomyces cerevisiae, Corynebacterium glutamicum, and Bacillus subtilis have been engineered to produce 2′-FL, their titers and productivities need be improved for economic production. Glucose along with lactose have been used as substrates for producing 2′-FL, but accumulation of by-products due to overflow metabolism of glucose hampered efficient production of 2′-FL regardless of a host strain. To circumvent this problem, we used xylose, which is the second most abundant sugar in plant cell wall hydrolysates and is metabolized through oxidativemore »« less
Data for High-Level β-Carotene Production from Xylose by Engineered Saccharomyces cerevisiae without Overexpression of a Truncated HMG1 (tHMG1)

Dataset Sun, Liang ; Atkinson, Christine A. ; Lee, Ye-Gi ; ...

β‐Carotene is a natural pigment and health‐promoting metabolite, and has been widely used in the nutraceutical, feed, and cosmetic industries. Here, we engineered a GRAS yeast Saccharomyces cerevisiae to produce β‐carotene from xylose, the second most abundant and inedible sugar component of lignocellulose biomass. Specifically, a β‐carotene biosynthetic pathway containing crtYB, crtI, and crtE from Xanthophyllomyces dendrorhous was introduced into a xylose‐fermenting S. cerevisiae. The resulting strain produced β‐carotene from xylose at a titer threefold higher than from glucose. Interestingly, overexpression of tHMG1, which has been reported as a critical genetic perturbation to enhance metabolic fluxes in the mevalonate pathwaymore »« less

Similar Records