Characterization of a New α-l-Arabinofuranosidase from Penicillium sp. LYG 0704, and their Application in Lignocelluloses Degradation

Abstract

A gene (arf) encoding an α-L: -arabinofuranosidase (ARF) that hydrolyzes arabinose substituted on xylan was isolated from Penicillium sp. The gene was predicted to encode 339 amino acid residues showing 71-75% homology to GH family 54. E. coli expressed ARF showed optimal activity at 50°C and pH 5-6 on wheat arabinoxylan. The hydrolysis activities on oat spelt xylan by ARF and xylanase were 1.67-fold higher than that of xylanase alone. The synergistic effects of ARF and commercial enzymes (xylanase and cellulase) on popping-pretreated rice straw were 1.15-1.51-fold higher amounts of sugars released in the [ARF + xylanase + cellulase] mixture than in the mixtures [ARF + xylanase], [ARF + cellulase], and [xylanase + cellulase]. Moreover, the liberation of arabinose by ARF was enhanced 2.1-2.9-fold in a reaction with xylanase and cellulase as compared with [xylanase + cellulase] and ARF alone.

Synergistic effects of 2A-mediated polyproteins on the production of lignocellulose degradation enzymes in tobacco plants

“…Ng et al (2011) reported that supplementing β-glucosidase from Taiwanese fungi into an enzyme mixture of Trichoderma reesei , which consisted of the main cellulase enzymes CBH1, CBHII, and EG2 (representing 60, 20, and 12% of the mixture, respectively), enhanced the efficiency of hydrolysis into fermentable sugars. Supplementation with a hemicellulase cocktail containing the side-chain degradation enzymes, endoglucanase, and β-glucosidase of a microbe-produced enzyme mixture of Trichoderma reesei or Trichoderma viride led to more efficient degradation of lignocellulosic biomass (Tabka et al , 2006; Lee et al , 2011). Therefore, 2A-mediated multiple enzyme expression could prove to be a powerful strategy in the enzymic hydrolysis of lignocellulosic biomass when supplemented with additional enzymes.…”

“…For exoglucanase preparation, E3 ( Cel6B ) from Thermomonospora fusca was subcloned into pCold I Expression vector (TaKaRa) for expression in Escherichia coli (Lee et al , 2011) and E6 ( Cel48A ) from Thermomonospora fusca , and the CBHI and CBHII genes from Aspergillus flavus were individually introduced into the vector pPICZα A for expression in Pichia pastoris . The recombinant plasmids were linearized by restriction with Pme 1 and transformed into P. pastoris -competent cells by electroporation.…”

“…Efficient conversion of lignocellulosic biomass to fermentable sugars by enzymic hydrolysis requires three cellulose-degrading enzymes (β-1,4-endoglucanase, β-1,4-exoglucanases, and β- d -glucosidase), two hemicellulose-digesting enzymes (xylanase and xylosidase), and other enzymes involved in debranching side chains (e.g., arabinofuranosidase, ferulic acid esterase, glucuronidase, and acetyl xylan esterase) (Lee at al., 2011; Margolles-Clark, 1996; Saha et al , 2003). Production of lignocellulose hydrolysis enzymes by conventional microbial fermentation methods is not cost-effective for the saccharification of lignocellulosic biomass.…”

Synergistic action of recombinant accessory hemicellulolytic and pectinolytic enzymes to Trichoderma reesei cellulase on rice straw degradation

“…However, only a limited number of studies on synergism of a-arabinofuranosidases on hydrolysis of lignocellulosic substrates have been reported. These included the enhancement of xylose recovery by arabinofuranosidase mutant from Pleutus ostreatus on hydrolysis of pretreated Arundo donax, corn cobs, and brewer's spent grains (Marcolongo et al, 2014), synergistic effects of Penicillium arabinofuranosidase on hydrolysis of rice straw by cellulase and xylanase (Lee et al, 2011), and the combination of a-arabinofuranosidase to mannanase and xylanase on hydrolysis of bagasse (Beukes and Pletschke, 2010). In addition to the action of hydrolytic enzymes, PET can also further increase the efficiency of the enzyme systems by de-esterification of the pectin component, which acts as the embedded matrix in the cell wall structure.…”

Production, Purification and Characterization of Hemicellulose from Penicillium janthinellum 3CHP

“…Optimum temperature for the enzyme was also found to be 50°C for P. purpurogenum (Ioannes et al 2000) and P. sp. LYG0704 (Lee et al 2011). …”