Enhanced reducing sugar production and extraction for Chlorella vulgaris in mixotrophic cultivation using high hydrostatic pressure processing and ultrasound

Although extraction of polysaccharides to convert reducing sugars (RS) from microalgae by acid or alkali pretreatments and enzymatic hydrolysis has been extensively studied, few reports exploring the use of high hydrostatic pressure processing (HHP) and ultrasonication (US) as emerging technologies for the extraction of sugars from microalgae biomass exist. Thus, the present study was conducted to determine the effects of mixotrophic growth and stress conditions (NaNO3 and CO2 concentration and light intensity) on RS and protein accumulation in the unicellular green alga Chlorella vulgaris in addition to optimization of the effectiveness of the sequential applications of HHP and US with dilute acid as well as simultaneous enzymatic saccharification on the production of RS from microalga cells. High light intensity, high CO2 concentration and limited nitrogen concentration promoted RS production. The maximum protein content (0.0683 mg g(-1)) was achieved at 0.3 g l(-1) NaNO3 concentration, 7000 mu mol photons m(-2) s(-1) and 6 l min(-1) CO2 concentration. The highest RS content of C. vulgaris after 48 h enzymatic saccharification (583.86 +/- 13.23 mg g(-1)) was obtained at 1% (w/w) acid concentration and 80% amplitude for 30 min with 79.4% RS yield. Combined US-assisted dilute acid pretreatment and enzymatic hydrolysis were also found to be more effective than HHP assisted dilute acid pretreatment and enzymatic saccharification. Therefore, microalgal biomass can be considered a suitable renewable feedstock used in fermentation. Highlights center dot The cultivation period of Chlorella vulgaris was reduced from 25 days to 14 days using mixotrophic growing conditions.center dot Mixotrophic conditions enhanced reducing sugar productivity.center dot Novel extraction techniques enhanced the extraction of reducing sugar from microalgae.