Scaling up and scaling down the production of galactaric acid from pectin using Trichoderma reesei

Research output: Contribution to journalArticleScientificpeer-review

4 Citations (Scopus)

Abstract

Background: Bioconversion of d-galacturonic acid to galactaric (mucic) acid has previously been carried out in small scale (50-1000 mL) cultures, which produce tens of grams of galactaric acid. To obtain larger amounts of biologically produced galactaric acid, the process needed to be scaled up using a readily available technical substrate. Food grade pectin was selected as a readily available source of d-galacturonic acid for conversion to galactaric acid. Results: We demonstrated that the process using Trichoderma reesei QM6a Δgar1 udh can be scaled up from 1 L to 10 and 250 L, replacing pure d-galacturonic acid with commercially available pectin. T. reesei produced 18 g L -1 galactaric acid from food-grade pectin (yield 1.00 g [g d-galacturonate consumed] -1) when grown at 1 L scale, 21 g L -1 galactaric acid (yield 1.11 g [g d-galacturonate consumed] -1) when grown at 10 L scale and 14 g L -1 galactaric acid (yield 0.77 g [g d-galacturonate consumed] -1) when grown at 250 L scale. Initial production rates were similar to those observed in 500 mL cultures with pure d-galacturonate as substrate. Approximately 2.8 kg galactaric acid was precipitated from the 250 L culture, representing a recovery of 77% of the galactaric acid in the supernatant. In addition to scaling up, we also demonstrated that the process could be scaled down to 4 mL for screening of production strains in 24-well plate format. Production of galactaric acid from pectin was assessed for three strains expressing uronate dehydrogenase under alternative promoters and up to 11 g L -1 galactaric acid were produced in the batch process. Conclusions: The process of producing galactaric acid by bioconversion with T. reesei was demonstrated to be equally efficient using pectin as it was with d-galacturonic acid. The 24-well plate batch process will be useful screening new constructs, but cannot replace process optimisation in bioreactors. Scaling up to 250 L demonstrated good reproducibility with the smaller scale but there was a loss in yield at 250 L which indicated that total biomass extraction and more efficient DSP would both be needed for a large scale process.

Original languageEnglish
Article number119
JournalMicrobial Cell Factories
Volume16
Issue number1
DOIs
Publication statusPublished - 11 Jul 2017
MoE publication typeA1 Journal article-refereed

Fingerprint

Trichoderma
Acids
Bioconversion
uronate dehydrogenase
pectin
galactaric acid
Screening
Food
Substrates
Bioreactors
Biomass

Keywords

  • galactaric acid
  • mucic acid
  • d-Galacturonic acid
  • pectin
  • trichoderma reesei
  • scale-up
  • scale-down

Cite this

@article{324f10785847463e8322d28bf3fdbd50,
title = "Scaling up and scaling down the production of galactaric acid from pectin using Trichoderma reesei",
abstract = "Background: Bioconversion of d-galacturonic acid to galactaric (mucic) acid has previously been carried out in small scale (50-1000 mL) cultures, which produce tens of grams of galactaric acid. To obtain larger amounts of biologically produced galactaric acid, the process needed to be scaled up using a readily available technical substrate. Food grade pectin was selected as a readily available source of d-galacturonic acid for conversion to galactaric acid. Results: We demonstrated that the process using Trichoderma reesei QM6a Δgar1 udh can be scaled up from 1 L to 10 and 250 L, replacing pure d-galacturonic acid with commercially available pectin. T. reesei produced 18 g L -1 galactaric acid from food-grade pectin (yield 1.00 g [g d-galacturonate consumed] -1) when grown at 1 L scale, 21 g L -1 galactaric acid (yield 1.11 g [g d-galacturonate consumed] -1) when grown at 10 L scale and 14 g L -1 galactaric acid (yield 0.77 g [g d-galacturonate consumed] -1) when grown at 250 L scale. Initial production rates were similar to those observed in 500 mL cultures with pure d-galacturonate as substrate. Approximately 2.8 kg galactaric acid was precipitated from the 250 L culture, representing a recovery of 77{\%} of the galactaric acid in the supernatant. In addition to scaling up, we also demonstrated that the process could be scaled down to 4 mL for screening of production strains in 24-well plate format. Production of galactaric acid from pectin was assessed for three strains expressing uronate dehydrogenase under alternative promoters and up to 11 g L -1 galactaric acid were produced in the batch process. Conclusions: The process of producing galactaric acid by bioconversion with T. reesei was demonstrated to be equally efficient using pectin as it was with d-galacturonic acid. The 24-well plate batch process will be useful screening new constructs, but cannot replace process optimisation in bioreactors. Scaling up to 250 L demonstrated good reproducibility with the smaller scale but there was a loss in yield at 250 L which indicated that total biomass extraction and more efficient DSP would both be needed for a large scale process.",
keywords = "galactaric acid, mucic acid, d-Galacturonic acid, pectin, trichoderma reesei, scale-up, scale-down",
author = "Toni Paasikallio and Anne Huuskonen and Marilyn Wiebe",
year = "2017",
month = "7",
day = "11",
doi = "10.1186/s12934-017-0736-3",
language = "English",
volume = "16",
journal = "Microbial Cell Factories",
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}

Scaling up and scaling down the production of galactaric acid from pectin using Trichoderma reesei. / Paasikallio, Toni; Huuskonen, Anne; Wiebe, Marilyn.

In: Microbial Cell Factories, Vol. 16, No. 1, 119, 11.07.2017.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Scaling up and scaling down the production of galactaric acid from pectin using Trichoderma reesei

AU - Paasikallio, Toni

AU - Huuskonen, Anne

AU - Wiebe, Marilyn

PY - 2017/7/11

Y1 - 2017/7/11

N2 - Background: Bioconversion of d-galacturonic acid to galactaric (mucic) acid has previously been carried out in small scale (50-1000 mL) cultures, which produce tens of grams of galactaric acid. To obtain larger amounts of biologically produced galactaric acid, the process needed to be scaled up using a readily available technical substrate. Food grade pectin was selected as a readily available source of d-galacturonic acid for conversion to galactaric acid. Results: We demonstrated that the process using Trichoderma reesei QM6a Δgar1 udh can be scaled up from 1 L to 10 and 250 L, replacing pure d-galacturonic acid with commercially available pectin. T. reesei produced 18 g L -1 galactaric acid from food-grade pectin (yield 1.00 g [g d-galacturonate consumed] -1) when grown at 1 L scale, 21 g L -1 galactaric acid (yield 1.11 g [g d-galacturonate consumed] -1) when grown at 10 L scale and 14 g L -1 galactaric acid (yield 0.77 g [g d-galacturonate consumed] -1) when grown at 250 L scale. Initial production rates were similar to those observed in 500 mL cultures with pure d-galacturonate as substrate. Approximately 2.8 kg galactaric acid was precipitated from the 250 L culture, representing a recovery of 77% of the galactaric acid in the supernatant. In addition to scaling up, we also demonstrated that the process could be scaled down to 4 mL for screening of production strains in 24-well plate format. Production of galactaric acid from pectin was assessed for three strains expressing uronate dehydrogenase under alternative promoters and up to 11 g L -1 galactaric acid were produced in the batch process. Conclusions: The process of producing galactaric acid by bioconversion with T. reesei was demonstrated to be equally efficient using pectin as it was with d-galacturonic acid. The 24-well plate batch process will be useful screening new constructs, but cannot replace process optimisation in bioreactors. Scaling up to 250 L demonstrated good reproducibility with the smaller scale but there was a loss in yield at 250 L which indicated that total biomass extraction and more efficient DSP would both be needed for a large scale process.

AB - Background: Bioconversion of d-galacturonic acid to galactaric (mucic) acid has previously been carried out in small scale (50-1000 mL) cultures, which produce tens of grams of galactaric acid. To obtain larger amounts of biologically produced galactaric acid, the process needed to be scaled up using a readily available technical substrate. Food grade pectin was selected as a readily available source of d-galacturonic acid for conversion to galactaric acid. Results: We demonstrated that the process using Trichoderma reesei QM6a Δgar1 udh can be scaled up from 1 L to 10 and 250 L, replacing pure d-galacturonic acid with commercially available pectin. T. reesei produced 18 g L -1 galactaric acid from food-grade pectin (yield 1.00 g [g d-galacturonate consumed] -1) when grown at 1 L scale, 21 g L -1 galactaric acid (yield 1.11 g [g d-galacturonate consumed] -1) when grown at 10 L scale and 14 g L -1 galactaric acid (yield 0.77 g [g d-galacturonate consumed] -1) when grown at 250 L scale. Initial production rates were similar to those observed in 500 mL cultures with pure d-galacturonate as substrate. Approximately 2.8 kg galactaric acid was precipitated from the 250 L culture, representing a recovery of 77% of the galactaric acid in the supernatant. In addition to scaling up, we also demonstrated that the process could be scaled down to 4 mL for screening of production strains in 24-well plate format. Production of galactaric acid from pectin was assessed for three strains expressing uronate dehydrogenase under alternative promoters and up to 11 g L -1 galactaric acid were produced in the batch process. Conclusions: The process of producing galactaric acid by bioconversion with T. reesei was demonstrated to be equally efficient using pectin as it was with d-galacturonic acid. The 24-well plate batch process will be useful screening new constructs, but cannot replace process optimisation in bioreactors. Scaling up to 250 L demonstrated good reproducibility with the smaller scale but there was a loss in yield at 250 L which indicated that total biomass extraction and more efficient DSP would both be needed for a large scale process.

KW - galactaric acid

KW - mucic acid

KW - d-Galacturonic acid

KW - pectin

KW - trichoderma reesei

KW - scale-up

KW - scale-down

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U2 - 10.1186/s12934-017-0736-3

DO - 10.1186/s12934-017-0736-3

M3 - Article

VL - 16

JO - Microbial Cell Factories

JF - Microbial Cell Factories

SN - 1475-2859

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ER -