Functional characterization of two GH27 ɑ-galactosidases from Penicillium parvum 4–14 and their differential capabilities upon plant biomass degradation

IF 2.4 3区化学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Carbohydrate Research Pub Date : 2025-02-15 DOI:10.1016/j.carres.2025.109428

Wei Wang , Jiarong Zhu , Yizhou Wang , Liangkun Long , Qunying Lin , Jing Wang , Shaojun Ding

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引用次数: 0

Abstract

Two new ɑ-galactosidases PpAgl27B and PpAgl27C from Penicillium parvum 4–14 were functionally investigated in this study. Based on the analysis of catalytic domain and phylogenetic tree, PpAgl27B (435 aa) and PpAgl27C (543 aa) belong to glycoside hydrolase (GH) 27 family. After expression in Pichia pastoris, the recombinant PpAgl27B and PpAgl27C showed the highest activities at pH 3.5 and 65 °C, or 4.0 and 45 °C, respectively. Using p-nitrophenyl-α-d-galactopyranoside (pNPGal) as substrate, the Michaelis constant were 0.90 mM for PpAgl27B and 2.54 mM for PpAgl27C. PpAgl27C had a low catalytic activity toward pNPGal and negligible activities on various natural substrates. Differently, PpAgl27B efficiently released galactose from the artificial substrate, rafﬁnose family oligosaccharides, or galactomannans. Hydrolysis of corn bran arabinoxylan (CBAX) 1 or 2 were conducted by PpAgl27B alone or in combination with the enzyme blend E_CBAX1. PpAgl27B released a small amount of galactose (1.7–3.0 mg/g) from the both substrates. Compared with the individual enzymes, the liberations of galactose, xylose and arabinose from the substrates were significantly enhanced by combing PpAgl27B and E_CBAX1. The degrees of synergy of the enzyme combination for the saccharification of CBAX1 or CBAX2 were 1.20 and 1.13, respectively. PpAgl27B showed promising potential for the valorization of galactose-rich feedstocks as well as CBAX.

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来源期刊

Carbohydrate Research 化学-生化与分子生物学

CiteScore

5.00

自引率

3.20%

发文量

183

审稿时长

3.6 weeks

期刊介绍： Carbohydrate Research publishes reports of original research in the following areas of carbohydrate science: action of enzymes, analytical chemistry, biochemistry (biosynthesis, degradation, structural and functional biochemistry, conformation, molecular recognition, enzyme mechanisms, carbohydrate-processing enzymes, including glycosidases and glycosyltransferases), chemical synthesis, isolation of natural products, physicochemical studies, reactions and their mechanisms, the study of structures and stereochemistry, and technological aspects. Papers on polysaccharides should have a "molecular" component; that is a paper on new or modified polysaccharides should include structural information and characterization in addition to the usual studies of rheological properties and the like. A paper on a new, naturally occurring polysaccharide should include structural information, defining monosaccharide components and linkage sequence. Papers devoted wholly or partly to X-ray crystallographic studies, or to computational aspects (molecular mechanics or molecular orbital calculations, simulations via molecular dynamics), will be considered if they meet certain criteria. For computational papers the requirements are that the methods used be specified in sufficient detail to permit replication of the results, and that the conclusions be shown to have relevance to experimental observations - the authors'' own data or data from the literature. Specific directions for the presentation of X-ray data are given below under Results and "discussion".