Carbohydrate-binding module assisted purification and immobilization of β-glucosidase onto cellulose and application in hydrolysis of soybean isoflavone glycosides

Identification and Immobilization of an Invertase With High Specific Activity and Sucrose Tolerance Ability of Gongronella sp. w5 for High Fructose Syrup Preparation

“…Meanwhile, no GspInv was bound to cellulose (Figure 6). CBMs such as CBM 24 from family 3 usually exhibit high absorption efficiencies toward cellulose because they usually have a flat hydrophobic binding surface comprising of aromatic residues (Boraston et al, 2004;Chang et al, 2018). The planar architecture of the binding sites is consistent with the flat surfaces of crystalline polysaccharides such as cellulose, facilitating their irreversible binding and reaching a plateau within a short time (Boraston et al, 2004).…”

“…GspInv was purified and immobilized onto cellulose through fusing a CBM 24 from Clostridium thermocellum (GenBank No. HF912724.1) to GspInv (Chang et al, 2018). In brief, the CBM 24 gene was codon-optimized in accordance with the K. pastoris codon bias and synthesized by Sangon Biotech (Shanghai, China).…”

“…In brief, the CBM 24 gene was codon-optimized in accordance with the K. pastoris codon bias and synthesized by Sangon Biotech (Shanghai, China). It was fused to the N-or C-terminus of GspInv by using overlap extension PCR (Chang et al, 2018). Given that CBM 24 contains three potential glycosylation sites at 11, 65, and 121 in the amino acid sequence, the amino acids at these sites were mutated to aspartic acid to obtain CBM 24 DG (DG stands for deglycosylation).…”

Preparation of immobilized/stabilized biocatalysts of β‐glucosidases from different sources: Importance of the support active groups and the immobilization protocol

“…In this work, we evaluated the possibility to immobilize, improve the stability, and reduce glucose inhibition of β‐glucosidases from two different commercial preparations, Pectinex Ultra SP‐L and Celluclast® 1.5L using agarose beads activated by either glyoxyl, epoxide, or vinylsulfone. β‐Glucosidases (β‐ d ‐glucopyranoside glucohydrolase; EC 3.2.1.21) are very important enzymes in diverse areas from energy production (hydrolyzing cellobiose to fermentable glucose) to food and feed technology, pharmaceutical, textile, and paper industries . Both preparations, Pectinex Ultra SP‐L and Celluclast® 1.5L, are composed mainly by pectinase and cellulase activities, respectively, and were previously characterized by their β‐glucosidase activities, showing good specific activity and glucose tolerance, which encouraged us to improve their properties by immobilization.…”

“…β-Glucosidases (β-D-glucopyranoside glucohydrolase; EC 3.2.1.21) are very important enzymes in diverse areas from energy production (hydrolyzing cellobiose to fermentable glucose) to food and feed technology, pharmaceutical, textile, and paper industries. [37][38][39][40] Both preparations, Pectinex Ultra SP-L and Celluclast ® 1.5L, are composed mainly by pectinase and cellulase activities, respectively, and were previously characterized by their β-glucosidase activities, showing good specific activity and glucose tolerance, 41 which encouraged us to improve their properties by immobilization. In Figure 1, we present a schematic representation of all immobilizations tested in our work, showing the reactions between enzyme and support, and blocking agent and support.…”