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One-step multi-enzyme pretreatment and biohydrogen production from

sp. biomass
Chlorella

2,3,*
2,3
1,2,3
Kawinda Sriyod , Alissara Reungsang , Pensri Plangklang

1 Graduate School, Khon Kaen University, Khon Kaen 40002, Thailand
2 Research Group for Development of Microbial Hydrogen Production Process from Biomass, Khon Kaen University,
Khon Kaen 40002, Thailand
3 Department of Biotechnology, Faculty of Technology, Khon Kaen University, Khon Kaen, 40002, Thailand
*Corresponding author: penspl@kku.ac.th
________________________________________________________________________________
Abstract

One-step multi-enzyme (OSME) pretreatment of Chlorella sp. biomass was statistically
optimized. Enzyme dosages of 380 units/g-VS Termamyl SC, 229.45 units/g-VS Dextrozyme GA,

and 34.13 filter paper units/g-VS Cellic CTec 2, and a hydrolysis time of 90 minutes were the
optimal conditions yielding a reducing sugar yield of 167.61 mg/g-VS, equivalent to 49.29%

carbohydrate solubilization. OSME pretreated biomass was used as a substrate for biohydrogen
production that was optimized under a feed/inoculum ratio of 3.0 and initial pH of 6.5. A
hydrogen yield of 43.16 mL-H /g-VS was obtained under optimal conditions, which was 39.63%
2
greater than that observed with non-treated biomass. The addition of nutrients in the form of
a basic anaerobic medium with or without a nitrogen source tended to decrease the efficiency

of hydrogen production from the hydrolyzed biomass. This work demonstrated OSME
hydrolysis as an alternative to the sequential hydrolysis processes for efficient conversion of

microalgae biomass to fermentable sugars and biohydrogen.

Keywords: 3 generation biomass; one-step enzyme pretreatment; biogas; biofuel
rd