Development of hemicellulolytic enzyme mixtures for plant biomass deconstruction on target biotechnological applications

Abstract

An essential step in the conversion of lignocellulosic biomass to ethanol and other biorefinery products is conversion of cell wall polysaccharides into fermentable sugars by enzymatic hydrolysis. The objective of the present study was to understand the mode of action of hemicellulolytic enzyme mixtures for pretreated sugarcane bagasse (PSB) deconstruction and wheat arabinoxylan (WA) hydrolysis on target biotechnological applications. In this study, five hemicellulolytic enzymes-two endo-1,4-xylanases (GH10 and GH11), two α-L-arabinofuranosidases (GH51 and GH54), and one β-xylosidase (GH43)-were submitted to combinatorial assays using the experimental design strategy, in order to analyze synergistic and antagonistic effects of enzyme interactions on biomass degradation. The xylooligosaccharides (XOSs) released from hydrolysis were analyzed by capillary electrophoresis and quantified by high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD). Based on this analysis, it was possible to define which enzymatic combinations favor xylose (X1) or XOS production and thus enable the development of target biotechnological applications. Our results demonstrate that if the objective is X1 production from WA, the best enzymatic combination is GH11 + GH54 + GH43, and for xylobiose (X2) production from WA, it is best to combine GH11 + GH51. However, if the goal is to produce XOS, the five enzymes used in WA hydrolysis are important, but for PSB hydrolysis, only GH11 is sufficient. If the final objective is bioethanol production, GH11 is responsible for hydrolyzing 64.3 % of hemicellulose from PSB. This work provides a basis for further studies on enzymatic mechanisms for XOS production, and the development of more efficient and less expensive enzymatic mixtures, targeting commercially viable lignocellulosic ethanol production and other biorefinery products.

Screening of a Novel Glycoside Hydrolase Family 51 α-L-Arabinofuranosidase from Paenibacillus polymyxa KF-1: Cloning, Expression, and Characterization

“…Hemicellulose is one of the key components in lignocellulose [6]. Hemicellulose is mainly degraded by endoxylanase, β-xylosidase, α-L-arabinofuranosidase, mannanase, α-glucuronidase, and acetylxylan esterase [6,7]. α-L-arabinofuranosidase (Abf, EC.…”

The Coptotermes gestroi aldo–keto reductase: a multipurpose enzyme for biorefinery applications

“…No hydrolysis was detected by Cg AKR-1 or NAPDH in the absence of endoglucanase. The DS for sugar release was calculated as follows: (g/L) of reducing sugar of ( Cg GH9 + enzyme)/(g/L) of reducing sugars of Cg GH9, as described by Goldbeck [86]. …”

A NOVEL Scytalidium SPECIES: UNDERSTAND THE CELLULOLYTIC SYSTEM FOR BIOMASS SACCHARIFICATION

“…Because microcrystalline cellulose is a substrate with predominant crystalline structure, it may likely induce the production of cellobiohydrolases, whose product is cellobiose, a substrate easily hydrolyzed by β-glucosidases in only one step (Zhang et al, 2007). According to Maeda et al (2011) and Goldbeck et al (2014) The equation coefficients are all significant at the 95 % confidence level. The hypothesis of lack-of-fit of the equation was rejected, as the p-value = 0.014 is lower than 0.05.…”