Biosugars 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
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
Limited resources of D ‐ mannose & MOSs Extraction from plants Aloe vera Cranberries Palm kernel Konjac
Spent coffee grounds (SCG): - Everywhere - Cheap/valueless - Harmfull
Containing a lot of valuable polysaccharides… Mannan Galactomannan Arabinogalactan Redgwell et al., 2006
Produces D-mannose and MOSs from SCG Delignification SCG-derived SCG polysaccharides Defatting Chemical composition analysis Short-term saccharification Supernatant Solid Freeze drying 1 st SCG-derived biosugar Long-term saccharification Supernatant Delignification: NaClO 4 (1%, w/w) + Acetic acid (1%, 2 nd SCG-derived biosugar v/w); 3 hours; 80 ᴼ C, stirring. Defatting: Soxhlet; hexane:biomass (3:1, v/w); 6 hours, 60 ᴼ C, refluxing. Fermentation Development of an integrated process to produce d ‐ mannose and bioethanol from pH 9, 9hr Nguyen et al. 2017 coffee residue waste. Bioresour Technol. Pervaporation 2017 Nov;244(Pt 1):1039 ‐ 1048. doi: 10.1016/j.biortech.2017.07.169. Epub 2017 Aug 3. Freeze drying Nguyen QA , Cho E, Trinh LTP, Jeong JS, Bae HJ. 3 rd SCG-derived biosugar Bioethanol
Chemical compositions 100 50 SCG (control) 80 40 SCG ‐ derived polysaccharides 60 30 40 20 20 10 0 0
Mannose 60 40 20 0
SCG SCG-derived polysaccharides 500 X 20,000 X 3291.6cm -1 -OH 1708.9 cm -1 2851.5cm -1 -C=O 2919.4cm -1 -C-H stretch SGC SGC-derived polysaccharides 3900 3400 2900 2400 1900 1400 900 400 cm -1
Comparison enzymatic hydrolysis of SCG (control) and SCG-derived polysaccharides under hydrolysis of in-house produced pectinase Total MSs Total MOSs 80 80 Control SCG-derived polysaccharides 60 60 mg/mL mg/mL 40 40 20 20 0 0 0 2 4 6 8 10 12 0 2 4 6 8 10 12 Time (hr) Time (hr) (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 MSs MOSs 120 120 Glu M-6 M-4 M-3 M-2 Total MOSs Gal 100 100 Man 80 80 Total MSs mg/mL mg/mL 60 60 40 40 20 20 0 0 0 2 4 6 8 10 12 14 16 18 20 22 24 0 2 4 6 8 10 12 14 16 18 20 22 24 Time (hr) Time (hr)
RPM column - MSs ROA column - MOSs 3 rd biosugar 2 nd biosugar 3 rd biosugar 2 nd biosugar 1 st biosugar 1 st biosugar Ara M-5/6 Glu Gal Man M-3 M-4 M-2 Standards Standards 0 5 10 15 20 25 30 35 40 0 5 10 15 20 25
Delignification Defatting In-put SCG SCG-derived polysaccharides SCG-Delignified Out-put NaClO4 (1%, w/w) + Soxhlet: 100 g DW Acetic acid (1%, v/w); 3 Hexane:biomass ~77 g DW ~61 g DW hrs; 80 ᴼ C, stirring. 3:1; 6 hrs; 60 ᴼ C. Short-term In-house produced pectinase saccharification: 6 hours – 4.1 mg/g biomass 1 st SCG-derived biosugar: Supernatant Solid Manno-oligosaccharides & monosaccharides Freeze ~43 g DW drying ~15.9 g DW Long-term 1. In-house produced pectinase – 8.2 saccharification: 24 hours mg/g biomass Man 2. β -glucosidase – 2.1 mg/g biomass 9.9 Supernatant M-2 Freeze 24.5 drying M-6 19.8 2 nd SCG-derived biosugar: M-4 Monosaccharides & manno-oligosaccharides 12.0 ~25.6 g DW Man 49.0 Fermentation Yeast; Other nutrients pH 9, 6hr Pervaporation Liquid nitrogen Freeze drying 3 rd SCG-derived biosugar: Bioethanol Mannose ~3.1 g DW ~18.2 g DW Man 74.1
Conclusion Coffee is not only for drink. It would be for Biosugars - one of game-changers in biorefinery technology. 9.4 g MOSs 100 g 3.1 g 18.6 g SCG EtOH MSs 14.1 g Mannose
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