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Valuable biosugars including manno-oligosaccharides and mannose from spent coffee grounds

Bioenergy Research Center – BRC Chonnam National University, Korea

Quynh Anh, NGUYEN

New values of Coffee would be Biosugars

We believe that coffee is more than a pleasure drink

25 mil. 48 bil. ≥ 10 – 20 X

Esquivel & Jiménez, 2012

What is Biosugars?

• Monosaccharides
• Oligosaccharides
• Polysaccharides

Extraction from plants

Palm kernel Cranberries Konjac Aloe vera

Limited resources of D‐mannose & MOSs

Spent coffee grounds (SCG):

• Everywhere
• Cheap/valueless
• Harmfull

Redgwell et al., 2006

Mannan Galactomannan Arabinogalactan Containing a lot of valuable polysaccharides…

SCG SCG-derived polysaccharides Solid Supernatant Fermentation 3rd SCG-derived biosugar Pervaporation Bioethanol

pH 9, 9hr

Delignification Defatting Supernatant

Freeze drying

Chemical composition analysis Short-term saccharification Long-term saccharification 2nd SCG-derived biosugar 1st SCG-derived biosugar

Freeze drying

• Delignification: NaClO4 (1%, w/w) + Acetic acid (1%,

v/w); 3 hours; 80ᴼC, stirring.

• Defatting: Soxhlet; hexane:biomass (3:1, v/w); 6 hours,

60ᴼC, refluxing.

Produces D-mannose and MOSs from SCG

Nguyen et al. 2017

Development of an integrated process to produce d‐mannose and bioethanol from coffee residue waste. Bioresour Technol. 2017 Nov;244(Pt 1):1039‐1048. doi: 10.1016/j.biortech.2017.07.169. Epub 2017 Aug 3. Nguyen QA, Cho E, Trinh LTP, Jeong JS, Bae HJ.

20 40 60 80 100

SCG (control) SCG‐derived polysaccharides

Chemical compositions

10 20 30 40 50

20 40 60

Mannose

500 X 20,000 X

SCG SCG-derived polysaccharides

400 900 1400 1900 2400 2900 3400 3900

cm-1

2919.4cm-1 2851.5cm-1

• C=O

1708.9 cm-1

• C-H stretch

3291.6cm-1

• OH

SGC SGC-derived polysaccharides

20 40 60 80 2 4 6 8 10 12 mg/mL Time (hr)

Total MSs

Control SCG-derived polysaccharides 20 40 60 80 2 4 6 8 10 12 mg/mL Time (hr)

Total MOSs

Comparison enzymatic hydrolysis of SCG (control) and SCG-derived polysaccharides under hydrolysis of in-house produced pectinase

(Results from other commercial enzymes were not presented)

Conversion yield and rate were significantly increased after pretreatment

Enzymatic hydrolysis of SCG-derived polysaccharides under hydrolysis of in-house produced pectinase

20 40 60 80 100 120 2 4 6 8 10 12 14 16 18 20 22 24 mg/mL Time (hr)

MSs

Glu Gal Man Total MSs

20 40 60 80 100 120 2 4 6 8 10 12 14 16 18 20 22 24 mg/mL Time (hr)

MOSs

M-6 M-4 M-3 M-2 Total MOSs

15 20 10 5 25 30 35 Man Gal Glu Ara Standards 1st biosugar 2nd biosugar 3rd biosugar

RPM column - MSs ROA column - MOSs

Standards 1st biosugar 2nd biosugar 3rd biosugar M-2 M-3 M-4 M-5/6 15 20 10 5 25 40

SCG SCG-derived polysaccharides 1st SCG-derived biosugar: Manno-oligosaccharides & monosaccharides Solid 2nd SCG-derived biosugar: Monosaccharides & manno-oligosaccharides Supernatant Fermentation 3rd SCG-derived biosugar: Mannose Pervaporation Bioethanol

pH 9, 6hr

SCG-Delignified 100 g DW ~77 g DW ~61 g DW Delignification Defatting Supernatant

Freeze drying

Long-term saccharification: 24 hours ~15.9 g DW

Freeze drying

~25.6 g DW ~43 g DW ~18.2 g DW ~3.1 g DW

Freeze drying NaClO4 (1%, w/w) + Acetic acid (1%, v/w); 3 hrs; 80ᴼC, stirring. Soxhlet: Hexane:biomass 3:1; 6 hrs; 60ᴼC.

• 1. In-house produced pectinase – 8.2

mg/g biomass

• 2. β-glucosidase – 2.1 mg/g biomass

In-house produced pectinase – 4.1 mg/g biomass Yeast; Other nutrients Liquid nitrogen

In-put Out-put Short-term saccharification: 6 hours Man 9.9 M-2 24.5 M-4 12.0 M-6 19.8 Man 49.0 Man 74.1

100 g SCG 9.4 g MOSs 18.6 g MSs 14.1 g Mannose 3.1 g EtOH Conclusion

Coffee is not only for drink. It would be for Biosugars - one of game-changers in biorefinery technology.