Engineering filamentous fungi for conversion of d-galacturonic acid to L-galactonic acid

Joosu Kuivanen, Dominik Mojzita, Yanming Wang, Satu Hilditch, Merja Penttilä, Peter Richard, Marilyn G. Wiebe (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

34 Citations (Scopus)

Abstract

D-Galacturonic acid, the main monomer of pectin, is an attractive substrate for bioconversions, since pectin-rich biomass is abundantly available and pectin is easily hydrolyzed. L-Galactonic acid is an intermediate in the eukaryotic pathway for D-galacturonic acid catabolism, but extracellular accumulation of L-galactonic acid has not been reported. By deleting the gene encoding L-galactonic acid dehydratase (lgd1 or gaaB) in two filamentous fungi, strains were obtained that converted D-galacturonic acid to L-galactonic acid. Both Trichoderma reesei Δlgd1 and Aspergillus niger ΔgaaB strains produced L-galactonate at yields of 0.6 to 0.9 g per g of substrate consumed. Although T. reesei Δlgd1 could produce L-galactonate at pH 5.5, a lower pH was necessary for A. niger ΔgaaB. Provision of a cosubstrate improved the production rate and titer in both strains. Intracellular accumulation of L-galactonate (40 to 70 mg g biomassΔ1) suggested that export may be limiting. Deletion of the L-galactonate dehydratase from A. niger was found to delay induction of D-galacturonate reductase and overexpression of the reductase improved initial production rates. Deletion of the L-galactonate dehydratase from A. niger also delayed or prevented induction of the putative D-galacturonate transporter An14g04280. In addition, A. niger ΔgaaB produced L-galactonate from polygalacturonate as efficiently as from the monomer.

Original languageEnglish
Pages (from-to)8676-8683
JournalApplied and Environmental Microbiology
Volume78
Issue number24
DOIs
Publication statusPublished - 1 Dec 2012
MoE publication typeA1 Journal article-refereed

Fingerprint

galacturonic acid
Aspergillus niger
galactonate dehydratase
engineering
Fungi
fungus
fungi
acids
acid
pectins
Trichoderma reesei
Hydro-Lyases
Trichoderma
biotransformation
Biomass
substrate
transporters
catabolism
Oxidoreductases
galactonic acid

Cite this

@article{5e6f7dd213614748b73c02e78ef9e911,
title = "Engineering filamentous fungi for conversion of d-galacturonic acid to L-galactonic acid",
abstract = "D-Galacturonic acid, the main monomer of pectin, is an attractive substrate for bioconversions, since pectin-rich biomass is abundantly available and pectin is easily hydrolyzed. L-Galactonic acid is an intermediate in the eukaryotic pathway for D-galacturonic acid catabolism, but extracellular accumulation of L-galactonic acid has not been reported. By deleting the gene encoding L-galactonic acid dehydratase (lgd1 or gaaB) in two filamentous fungi, strains were obtained that converted D-galacturonic acid to L-galactonic acid. Both Trichoderma reesei Δlgd1 and Aspergillus niger ΔgaaB strains produced L-galactonate at yields of 0.6 to 0.9 g per g of substrate consumed. Although T. reesei Δlgd1 could produce L-galactonate at pH 5.5, a lower pH was necessary for A. niger ΔgaaB. Provision of a cosubstrate improved the production rate and titer in both strains. Intracellular accumulation of L-galactonate (40 to 70 mg g biomassΔ1) suggested that export may be limiting. Deletion of the L-galactonate dehydratase from A. niger was found to delay induction of D-galacturonate reductase and overexpression of the reductase improved initial production rates. Deletion of the L-galactonate dehydratase from A. niger also delayed or prevented induction of the putative D-galacturonate transporter An14g04280. In addition, A. niger ΔgaaB produced L-galactonate from polygalacturonate as efficiently as from the monomer.",
author = "Joosu Kuivanen and Dominik Mojzita and Yanming Wang and Satu Hilditch and Merja Penttil{\"a} and Peter Richard and Wiebe, {Marilyn G.}",
note = "CA2: TK402 CA2: TK400 SDA: BIC ISI: BIOTECHNOLOGY & APPLIED MICROBIOLOGY",
year = "2012",
month = "12",
day = "1",
doi = "10.1128/AEM.02171-12",
language = "English",
volume = "78",
pages = "8676--8683",
journal = "Applied and Environmental Microbiology",
issn = "0099-2240",
publisher = "American Society for Microbiology",
number = "24",

}

Engineering filamentous fungi for conversion of d-galacturonic acid to L-galactonic acid. / Kuivanen, Joosu; Mojzita, Dominik; Wang, Yanming; Hilditch, Satu; Penttilä, Merja; Richard, Peter; Wiebe, Marilyn G. (Corresponding Author).

In: Applied and Environmental Microbiology, Vol. 78, No. 24, 01.12.2012, p. 8676-8683.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Engineering filamentous fungi for conversion of d-galacturonic acid to L-galactonic acid

AU - Kuivanen, Joosu

AU - Mojzita, Dominik

AU - Wang, Yanming

AU - Hilditch, Satu

AU - Penttilä, Merja

AU - Richard, Peter

AU - Wiebe, Marilyn G.

N1 - CA2: TK402 CA2: TK400 SDA: BIC ISI: BIOTECHNOLOGY & APPLIED MICROBIOLOGY

PY - 2012/12/1

Y1 - 2012/12/1

N2 - D-Galacturonic acid, the main monomer of pectin, is an attractive substrate for bioconversions, since pectin-rich biomass is abundantly available and pectin is easily hydrolyzed. L-Galactonic acid is an intermediate in the eukaryotic pathway for D-galacturonic acid catabolism, but extracellular accumulation of L-galactonic acid has not been reported. By deleting the gene encoding L-galactonic acid dehydratase (lgd1 or gaaB) in two filamentous fungi, strains were obtained that converted D-galacturonic acid to L-galactonic acid. Both Trichoderma reesei Δlgd1 and Aspergillus niger ΔgaaB strains produced L-galactonate at yields of 0.6 to 0.9 g per g of substrate consumed. Although T. reesei Δlgd1 could produce L-galactonate at pH 5.5, a lower pH was necessary for A. niger ΔgaaB. Provision of a cosubstrate improved the production rate and titer in both strains. Intracellular accumulation of L-galactonate (40 to 70 mg g biomassΔ1) suggested that export may be limiting. Deletion of the L-galactonate dehydratase from A. niger was found to delay induction of D-galacturonate reductase and overexpression of the reductase improved initial production rates. Deletion of the L-galactonate dehydratase from A. niger also delayed or prevented induction of the putative D-galacturonate transporter An14g04280. In addition, A. niger ΔgaaB produced L-galactonate from polygalacturonate as efficiently as from the monomer.

AB - D-Galacturonic acid, the main monomer of pectin, is an attractive substrate for bioconversions, since pectin-rich biomass is abundantly available and pectin is easily hydrolyzed. L-Galactonic acid is an intermediate in the eukaryotic pathway for D-galacturonic acid catabolism, but extracellular accumulation of L-galactonic acid has not been reported. By deleting the gene encoding L-galactonic acid dehydratase (lgd1 or gaaB) in two filamentous fungi, strains were obtained that converted D-galacturonic acid to L-galactonic acid. Both Trichoderma reesei Δlgd1 and Aspergillus niger ΔgaaB strains produced L-galactonate at yields of 0.6 to 0.9 g per g of substrate consumed. Although T. reesei Δlgd1 could produce L-galactonate at pH 5.5, a lower pH was necessary for A. niger ΔgaaB. Provision of a cosubstrate improved the production rate and titer in both strains. Intracellular accumulation of L-galactonate (40 to 70 mg g biomassΔ1) suggested that export may be limiting. Deletion of the L-galactonate dehydratase from A. niger was found to delay induction of D-galacturonate reductase and overexpression of the reductase improved initial production rates. Deletion of the L-galactonate dehydratase from A. niger also delayed or prevented induction of the putative D-galacturonate transporter An14g04280. In addition, A. niger ΔgaaB produced L-galactonate from polygalacturonate as efficiently as from the monomer.

UR - http://www.scopus.com/inward/record.url?scp=84871030777&partnerID=8YFLogxK

U2 - 10.1128/AEM.02171-12

DO - 10.1128/AEM.02171-12

M3 - Article

C2 - 23042175

AN - SCOPUS:84871030777

VL - 78

SP - 8676

EP - 8683

JO - Applied and Environmental Microbiology

JF - Applied and Environmental Microbiology

SN - 0099-2240

IS - 24

ER -