**E**ff**ect of Novel** *Penicillium verruculosum* **Enzyme Preparations on the Saccharification of Acid- and Alkali-Pretreated Agro-Industrial Residues**

**Susan G. Karp <sup>1</sup> , Dmitrii O. Osipov <sup>2</sup> , Margarita V. Semenova <sup>2</sup> , Alexandra M. Rozhkova 2,\* , Ivan N. Zorov 2,3, Olga A. Sinitsyna <sup>3</sup> , Carlos R. Soccol <sup>1</sup> and Arkady P. Sinitsyn 2,3**


Received: 18 August 2020; Accepted: 4 September 2020; Published: 7 September 2020

**Abstract:** This study aimed at evaluating different enzyme combinations in the saccharification of sugarcane bagasse (SCB), soybean husks (SBH) and oil palm empty fruit bunches (EFB) submitted to mild acid and alkaline pretreatments. Enzyme pools were represented by B1 host (crude cellulase/xylanase complexes of *Penicillium verruculosum*); B1-XylA (*Penicillium canescens* xylanase A expressed in *P. verruculosum* B1 host strain); and F10 (*Aspergillus niger* β-glucosidase expressed in B1 host strain). Enzyme loading was 10 mg protein/g dry substrate and 40 U/g of β-glucosidase (F10) activity. SCB was efficiently hydrolyzed by B1 host after alkaline pretreatment, yielding glucose and reducing sugars at 71 g/L or 65 g/100 g of dry pretreated substrate and 91 g/L or 83 g/100 g, respectively. B1 host performed better also for EFB, regardless of the pretreatment method, but yields were lower (glucose 27–30 g/L, 25–27 g/100 g; reducing sugars 37–42 g/L, 34–38 g/100 g). SBH was efficiently saccharified by the combination of B1 host and B1-XylA, yielding similar concentrations of reducing sugars for both pretreatments (92–96 g/L, 84–87 g/100 g); glucose recovery, however, was higher with alkaline pretreatment (81 g/L, 74 g/100 g). Glucose and reducing sugar yields from initial substrate mass were 42% and 54% for SCB, 36% and 42–47% for SBH and 16–18% and 21–26% for EFB, respectively.

**Keywords:** sugarcane bagasse; soybean husks; palm empty fruit bunches; recombinant enzymes; pretreatment
