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首页> 美国卫生研究院文献>Applied and Environmental Microbiology >The Wood Rot Ascomycete Xylaria polymorpha Produces a Novel GH78 Glycoside Hydrolase That Exhibits α-l-Rhamnosidase and Feruloyl Esterase Activities and Releases Hydroxycinnamic Acids from Lignocelluloses
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The Wood Rot Ascomycete Xylaria polymorpha Produces a Novel GH78 Glycoside Hydrolase That Exhibits α-l-Rhamnosidase and Feruloyl Esterase Activities and Releases Hydroxycinnamic Acids from Lignocelluloses

机译：木材腐烂的子囊菌Xylaria polymorpha生产出一种新型的GH78糖苷水解酶该酶表现出α-1-鼠李糖苷酶和阿魏酸酯酶的活性并从木质纤维素中释放出羟基肉桂酸。

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Soft rot (type II) fungi belonging to the family Xylariaceae are known to substantially degrade hardwood by means of their poorly understood lignocellulolytic system, which comprises various hydrolases, including feruloyl esterases and laccase. In the present study, several members of the Xylariaceae were found to exhibit high feruloyl esterase activity during growth on lignocellulosic materials such as wheat straw (up to 1,675 mU g−1) or beech wood (up to 80 mU g−1). Following the ester-cleaving activity toward methyl ferulate, a hydrolase of Xylaria polymorpha was produced in solid-state culture on wheat straw and purified by different steps of anion-exchange and size-exclusion chromatography to apparent homogeneity (specific activity, 2.2 U mg−1). The peptide sequence of the purified protein deduced from the gene sequence and verified by de novo peptide sequencing shows high similarity to putative α-l-rhamnosidase sequences belonging to the glycoside hydrolase family 78 (GH78; classified under EC 3.2.1.40). The purified enzyme (98 kDa by SDS-PAGE, 103 kDa by size-exclusion chromatography; pI 3.7) converted diverse glycosides (e.g., α-l-rhamnopyranoside and α-l-arabinofuranoside) but also natural and synthetic esters (e.g., chlorogenic acid, hydroxycinnamic acid glycoside esters, veratric acid esters, or p-nitrophenyl acetate) and released free hydroxycinnamic acids (ferulic and coumaric acid) from arabinoxylan and milled wheat straw. These catalytic properties strongly suggest that X. polymorpha GH78 is a multifunctional enzyme. It is the first fungal enzyme that combines glycosyl hydrolase with esterase activities and may help this soft rot fungus to degrade lignocelluloses.
机译：众所周知，属于木糖科的软腐（II型）真菌通过对木质纤维素分解系统的了解不多，可以显着降解硬木，其中包括各种水解酶，包括阿魏酸酯酶和漆酶。在本研究中，发现木糖科的几个成员在木质纤维素材料（例如麦秸（高达1,675 mU g -1 ）或山毛榉木（高达80）上生长期间表现出高的阿魏酸酯酶活性。 mU g -1 ）。继具有对阿魏酸甲酯的酯裂解活性之后，在小麦秸秆上的固态培养中产生了多形木瓜水解酶，并通过阴离子交换和尺寸排阻色谱的不同步骤纯化至表观同质性（比活性，2.2 U mg < sup> -1 ）。从基因序列推导并通过从头肽测序证实的纯化蛋白的肽序列显示出与属于糖苷水解酶家族78（GH78；归类于EC 3.2.1.40）的推定的α-1-鼠李糖苷酶序列的高度相似性。纯化的酶（通过SDS-PAGE为98 kDa，通过大小排阻色谱法为103 kDa； pI 3.7）不仅可以转化各种糖苷（例如α-1-鼠李糖吡喃糖苷和α-1-阿拉伯呋喃糖苷），还可以转化天然酯和合成酯（例如绿原酸酯）酸，羟基肉桂酸糖苷酯，维酸酯或对硝基苯乙酸酯），并从阿拉伯木聚糖和磨过的麦草中释放出游离的羟基肉桂酸（阿魏酸和香豆酸）。这些催化性能强烈表明多形木霉GH78是一种多功能酶。它是第一个结合糖基水解酶和酯酶活性的真菌酶，可以帮助这种软腐真菌降解木质纤维素。

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