Construction of a cell-free synthetic pathway for the production of lactic acid from spent coffee grounds

D. Kopp, P. L. Bergquist, R. Willows, A. Sunna

Research output: Contribution to journalMeeting abstractResearch

Abstract

Coffee is the second largest traded commodity after petroleum but almost half of it ends up as waste in form of spent coffee grounds. Over 50% of the spent coffee grounds are composed of carbohydrates, with mannose representing the most abundant sugar. However, a holistic approach for its efficient utilisation has not been addressed yet. Cell-free synthetic biology is developing into a platform technology enabling a rapid construction of enzymatic pathways for the production of platform chemicals and pharmaceuticals from various substrates. Here, we designed a cell-free synthetic pathway for the utilisation of mannose derived from spent coffee grounds to produce the platform chemical lactic acid. The concept for the synthetic pathway is based on a putative mannose metabolic pathway from the thermoacidophilic archaeon Thermoplasma acidophilum. We were able to reconstruct this metabolic route by heterologous overexpression of mannose dehydrogenase and mannonate dehydratase genes, and successfully convert mannose into 2-keto-3-deoxygluconate (2-KDG) in a one-pot enzymatic reaction. Further addition of two more enzymes resulted in a conversion of 2-KDG into lactic acid in a controlled and stereo-selective manner. Solid binding peptides (SBP) show great binding capacity towards a wide range of solid materials for the directed immobilisation of proteins and enzymes on solid supports. By co-expression of a silica-specific SBP, the pathway enzymes can be immobilised onto cheap silica-based supports like zeolite. This creates a range of reusable biocatalatyic modules, which can be rapidly assembled for future constructions of cell-free production pathways.
LanguageEnglish
Article numberP1-6
PagesS69-S70
Number of pages2
JournalNew Biotechnology
Volume44
Issue numberSupplement
DOIs
Publication statusPublished - 10 Oct 2018
Event18th European Congress on Biotechnology - Geneva, Switzerland
Duration: 1 Jul 20184 Jul 2018

Fingerprint

Artificial Cells
Coffee
Lactic acid
Mannose
Lactic Acid
Enzymes
Catalyst supports
Peptides
Silicon Dioxide
Silica
Thermoplasma
Synthetic Biology
Zeolites
Carbohydrates
Sugars
Drug products
Archaea
Petroleum
Metabolic Networks and Pathways
Crude oil

Keywords

  • Waste
  • coffee
  • lactic acid
  • synthetic pathway

Cite this

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title = "Construction of a cell-free synthetic pathway for the production of lactic acid from spent coffee grounds",
abstract = "Coffee is the second largest traded commodity after petroleum but almost half of it ends up as waste in form of spent coffee grounds. Over 50{\%} of the spent coffee grounds are composed of carbohydrates, with mannose representing the most abundant sugar. However, a holistic approach for its efficient utilisation has not been addressed yet. Cell-free synthetic biology is developing into a platform technology enabling a rapid construction of enzymatic pathways for the production of platform chemicals and pharmaceuticals from various substrates. Here, we designed a cell-free synthetic pathway for the utilisation of mannose derived from spent coffee grounds to produce the platform chemical lactic acid. The concept for the synthetic pathway is based on a putative mannose metabolic pathway from the thermoacidophilic archaeon Thermoplasma acidophilum. We were able to reconstruct this metabolic route by heterologous overexpression of mannose dehydrogenase and mannonate dehydratase genes, and successfully convert mannose into 2-keto-3-deoxygluconate (2-KDG) in a one-pot enzymatic reaction. Further addition of two more enzymes resulted in a conversion of 2-KDG into lactic acid in a controlled and stereo-selective manner. Solid binding peptides (SBP) show great binding capacity towards a wide range of solid materials for the directed immobilisation of proteins and enzymes on solid supports. By co-expression of a silica-specific SBP, the pathway enzymes can be immobilised onto cheap silica-based supports like zeolite. This creates a range of reusable biocatalatyic modules, which can be rapidly assembled for future constructions of cell-free production pathways.",
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Construction of a cell-free synthetic pathway for the production of lactic acid from spent coffee grounds. / Kopp, D.; Bergquist, P. L.; Willows, R.; Sunna, A.

In: New Biotechnology, Vol. 44, No. Supplement, P1-6, 10.10.2018, p. S69-S70.

Research output: Contribution to journalMeeting abstractResearch

TY - JOUR

T1 - Construction of a cell-free synthetic pathway for the production of lactic acid from spent coffee grounds

AU - Kopp, D.

AU - Bergquist, P. L.

AU - Willows, R.

AU - Sunna, A.

PY - 2018/10/10

Y1 - 2018/10/10

N2 - Coffee is the second largest traded commodity after petroleum but almost half of it ends up as waste in form of spent coffee grounds. Over 50% of the spent coffee grounds are composed of carbohydrates, with mannose representing the most abundant sugar. However, a holistic approach for its efficient utilisation has not been addressed yet. Cell-free synthetic biology is developing into a platform technology enabling a rapid construction of enzymatic pathways for the production of platform chemicals and pharmaceuticals from various substrates. Here, we designed a cell-free synthetic pathway for the utilisation of mannose derived from spent coffee grounds to produce the platform chemical lactic acid. The concept for the synthetic pathway is based on a putative mannose metabolic pathway from the thermoacidophilic archaeon Thermoplasma acidophilum. We were able to reconstruct this metabolic route by heterologous overexpression of mannose dehydrogenase and mannonate dehydratase genes, and successfully convert mannose into 2-keto-3-deoxygluconate (2-KDG) in a one-pot enzymatic reaction. Further addition of two more enzymes resulted in a conversion of 2-KDG into lactic acid in a controlled and stereo-selective manner. Solid binding peptides (SBP) show great binding capacity towards a wide range of solid materials for the directed immobilisation of proteins and enzymes on solid supports. By co-expression of a silica-specific SBP, the pathway enzymes can be immobilised onto cheap silica-based supports like zeolite. This creates a range of reusable biocatalatyic modules, which can be rapidly assembled for future constructions of cell-free production pathways.

AB - Coffee is the second largest traded commodity after petroleum but almost half of it ends up as waste in form of spent coffee grounds. Over 50% of the spent coffee grounds are composed of carbohydrates, with mannose representing the most abundant sugar. However, a holistic approach for its efficient utilisation has not been addressed yet. Cell-free synthetic biology is developing into a platform technology enabling a rapid construction of enzymatic pathways for the production of platform chemicals and pharmaceuticals from various substrates. Here, we designed a cell-free synthetic pathway for the utilisation of mannose derived from spent coffee grounds to produce the platform chemical lactic acid. The concept for the synthetic pathway is based on a putative mannose metabolic pathway from the thermoacidophilic archaeon Thermoplasma acidophilum. We were able to reconstruct this metabolic route by heterologous overexpression of mannose dehydrogenase and mannonate dehydratase genes, and successfully convert mannose into 2-keto-3-deoxygluconate (2-KDG) in a one-pot enzymatic reaction. Further addition of two more enzymes resulted in a conversion of 2-KDG into lactic acid in a controlled and stereo-selective manner. Solid binding peptides (SBP) show great binding capacity towards a wide range of solid materials for the directed immobilisation of proteins and enzymes on solid supports. By co-expression of a silica-specific SBP, the pathway enzymes can be immobilised onto cheap silica-based supports like zeolite. This creates a range of reusable biocatalatyic modules, which can be rapidly assembled for future constructions of cell-free production pathways.

KW - Waste

KW - coffee

KW - lactic acid

KW - synthetic pathway

U2 - 10.1016/j.nbt.2018.05.872

DO - 10.1016/j.nbt.2018.05.872

M3 - Meeting abstract

VL - 44

SP - S69-S70

JO - New Biotechnology

T2 - New Biotechnology

JF - New Biotechnology

SN - 1871-6784

IS - Supplement

M1 - P1-6

ER -