skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Novel Bioengineered Cassava Expressing an Archaeal Starch Degradation System and a Bacterial ADP-Glucose Pyrophosphorylase for Starch Self-Digestibility and Yield Increase

Abstract

To address national and global low-carbon fuel targets, there is great interest in alternative plant species such as cassava ( Manihot esculenta), which are high-yielding, resilient, and are easily converted to fuels using the existing technology. In this study the genes encoding hyperthermophilic archaeal starch-hydrolyzing enzymes, α-amylase and amylopullulanase from Pyrococcus furiosus and glucoamylase from Sulfolobus solfataricus, together with the gene encoding a modified ADP-glucose pyrophosphorylase ( glgC) from Escherichia coli, were simultaneously expressed in cassava roots to enhance starch accumulation and its subsequent hydrolysis to sugar. A total of 13 multigene expressing transgenic lines were generated and characterized phenotypically and genotypically. Gene expression analysis using quantitative RT-PCR showed that the microbial genes are expressed in the transgenic roots. Multigene-expressing transgenic lines produced up to 60% more storage root yield than the non-transgenic control, likely due to glgC expression. Total protein extracted from the transgenic roots showed up to 10-fold higher starch-degrading activity in vitro than the protein extracted from the non-transgenic control. Interestingly, transgenic tubers released threefold more glucose than the non-transgenic control when incubated at 85°C for 21-h without exogenous application of thermostable enzymes, suggesting that the archaeal enzymes produced in planta maintain their activity and thermostability.

Authors:
 [1];  [1];  [1];  [2];  [2];  [2];  [3];  [2];  [1]
  1. Delaware State Univ., Dover, DE (United States)
  2. Univ. of Georgia, Athens, GA (United States)
  3. Michigan State Univ., East Lansing, MI (United States)
Publication Date:
Research Org.:
Univ. of Georgia, Athens, GA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1510781
Grant/Contract Number:  
FG05-95ER20175
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Frontiers in Plant Science
Additional Journal Information:
Journal Volume: 9; Journal ID: ISSN 1664-462X
Publisher:
Frontiers Research Foundation
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; cassava; hyperthermophilic archaeal; glgC; starch self-processing; bioethanol; multigene-expression; starch-hydrolyzing enzymes

Citation Formats

Ligaba-Osena, Ayalew, Jones, Jenna, Donkor, Emmanuel, Chandrayan, Sanjeev, Pole, Farris, Wu, Chang -Hao, Vieille, Claire, Adams, Michael W. W., and Hankoua, Bertrand B. Novel Bioengineered Cassava Expressing an Archaeal Starch Degradation System and a Bacterial ADP-Glucose Pyrophosphorylase for Starch Self-Digestibility and Yield Increase. United States: N. p., 2018. Web. doi:10.3389/fpls.2018.00192.
Ligaba-Osena, Ayalew, Jones, Jenna, Donkor, Emmanuel, Chandrayan, Sanjeev, Pole, Farris, Wu, Chang -Hao, Vieille, Claire, Adams, Michael W. W., & Hankoua, Bertrand B. Novel Bioengineered Cassava Expressing an Archaeal Starch Degradation System and a Bacterial ADP-Glucose Pyrophosphorylase for Starch Self-Digestibility and Yield Increase. United States. doi:10.3389/fpls.2018.00192.
Ligaba-Osena, Ayalew, Jones, Jenna, Donkor, Emmanuel, Chandrayan, Sanjeev, Pole, Farris, Wu, Chang -Hao, Vieille, Claire, Adams, Michael W. W., and Hankoua, Bertrand B. Mon . "Novel Bioengineered Cassava Expressing an Archaeal Starch Degradation System and a Bacterial ADP-Glucose Pyrophosphorylase for Starch Self-Digestibility and Yield Increase". United States. doi:10.3389/fpls.2018.00192. https://www.osti.gov/servlets/purl/1510781.
@article{osti_1510781,
title = {Novel Bioengineered Cassava Expressing an Archaeal Starch Degradation System and a Bacterial ADP-Glucose Pyrophosphorylase for Starch Self-Digestibility and Yield Increase},
author = {Ligaba-Osena, Ayalew and Jones, Jenna and Donkor, Emmanuel and Chandrayan, Sanjeev and Pole, Farris and Wu, Chang -Hao and Vieille, Claire and Adams, Michael W. W. and Hankoua, Bertrand B.},
abstractNote = {To address national and global low-carbon fuel targets, there is great interest in alternative plant species such as cassava (Manihot esculenta), which are high-yielding, resilient, and are easily converted to fuels using the existing technology. In this study the genes encoding hyperthermophilic archaeal starch-hydrolyzing enzymes, α-amylase and amylopullulanase from Pyrococcus furiosus and glucoamylase from Sulfolobus solfataricus, together with the gene encoding a modified ADP-glucose pyrophosphorylase (glgC) from Escherichia coli, were simultaneously expressed in cassava roots to enhance starch accumulation and its subsequent hydrolysis to sugar. A total of 13 multigene expressing transgenic lines were generated and characterized phenotypically and genotypically. Gene expression analysis using quantitative RT-PCR showed that the microbial genes are expressed in the transgenic roots. Multigene-expressing transgenic lines produced up to 60% more storage root yield than the non-transgenic control, likely due to glgC expression. Total protein extracted from the transgenic roots showed up to 10-fold higher starch-degrading activity in vitro than the protein extracted from the non-transgenic control. Interestingly, transgenic tubers released threefold more glucose than the non-transgenic control when incubated at 85°C for 21-h without exogenous application of thermostable enzymes, suggesting that the archaeal enzymes produced in planta maintain their activity and thermostability.},
doi = {10.3389/fpls.2018.00192},
journal = {Frontiers in Plant Science},
issn = {1664-462X},
number = ,
volume = 9,
place = {United States},
year = {2018},
month = {2}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Save / Share:

Works referenced in this record:

Efficient Succinic Acid Production from Glucose through Overexpression of Pyruvate Carboxylase in an Escherichia coli Alcohol Dehydrogenase and Lactate Dehydrogenase Mutant
journal, January 2005

  • S�nchez, Ailen M.; Bennett, George N.; San, Ka-Yiu
  • Biotechnology Progress, Vol. 21, Issue 2, p. 358-365
  • DOI: 10.1021/bp049676e

Transcriptional and Biochemical Analysis of Starch Metabolism in the Hyperthermophilic Archaeon Pyrococcus furiosus
journal, March 2006


Autohydrolysis of plant polysaccharides using transgenic hyperthermophilic enzymes
journal, August 2000


Medium and techniques for induction and growth of monocotyledonous and dicotyledonous plant cell cultures
journal, January 1972

  • Schenk, Roy U.; Hildebrandt, A. C.
  • Canadian Journal of Botany, Vol. 50, Issue 1, p. 199-204
  • DOI: 10.1139/b72-026

A Revised Medium for Rapid Growth and Bio Assays with Tobacco Tissue Cultures
journal, July 1962


Engineering the Provitamin A (-Carotene) Biosynthetic Pathway into (Carotenoid-Free) Rice Endosperm
journal, January 2000


Industrial relevance of thermophilic Archaea
journal, December 2005

  • Egorova, Ksenia; Antranikian, Garabed
  • Current Opinion in Microbiology, Vol. 8, Issue 6, p. 649-655
  • DOI: 10.1016/j.mib.2005.10.015

Genome Sequencing of a Genetically Tractable Pyrococcus furiosus Strain Reveals a Highly Dynamic Genome
journal, May 2012

  • Bridger, S. L.; Lancaster, W. A.; Poole, F. L.
  • Journal of Bacteriology, Vol. 194, Issue 15, p. 4097-4106
  • DOI: 10.1128/JB.00439-12