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  • The intensification of amyloglucosidase-based saccharification by ultrasound.

The intensification of amyloglucosidase-based saccharification by ultrasound.

Ultrasonics sonochemistry (2018-08-14)
Hugo M Oliveira, Alice Q Pinheiro, António J M Fonseca, Ana R J Cabrita, Margarida R G Maia
ABSTRACT

The present report studied the role of ultrasound (US) energy in the amyloglucosidase-based starch hydrolysis using two complementary approaches: (i) in the activity of six commercially-available amyloglucosidases (using soluble starch as substrate), and (ii) in the hydrolysis of four pure starches from different botanical sources. This corresponds to the first systematic evaluation of the role of US in starch hydrolysis mediated by amyloglucosidase, being a consequence of our previous report that assessed the effect of US in the activity of alpha-amylase (LWT - Food Science and Technology 84 (2017) 674-685). Regarding amlyloglucosidases, three enzymes obtained from Aspergillus niger (AN1-AN3), and Spirizyme Achieve (SPA), Spirizyme Fuel (SPF) and Spirizyme Ultra (SPU) were submitted to a Box-Behnken experimental design in order to establish the optimum conditions for their maximum activity. In the presence of US, we found both inactivation and activation, ranging from -88% (AN3) to 699% (SPA). The US promoted the enzyme activity when combined with lower temperatures (40-60 °C), with a marked effect in Spirizyme enzymes. Based on the optimum conditions established by the experimental design, we also evaluated the role of US in the glucose yield resulting from the hydrolysis of pure starches (corn, rice, potato, wheat). In this case, US led to higher glucose yields in all conditions tested. The enhancement factors observed ranged from 1.2 (AN1, rice starch) to 65 (SPA, potato starch) times. We compared these findings with previous reports, which highlighted the role of US in intensifying amyloglucosidase-based saccharification in mild conditions, by simultaneously influencing both enzyme and substrate. Hence, US power has to be fine-tuned for each particular enzyme in order to maximize process intensification.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Starch from potato, Powder
Sigma-Aldrich
Starch from wheat, Unmodified
Supelco
Amyloglucosidase solution from Aspergillus niger, for use in Total Dietary Fiber Assay, TDF-100A
Supelco
Vials, screw top, silane-treated, volume 4 mL, amber glass vial, O.D. × H × I.D. 15 mm × 45 mm × 8 mm, thread for 13-425
Sigma-Aldrich
Starch, soluble, ACS reagent
Sigma-Aldrich
Starch from corn
Sigma-Aldrich
Starch from rice