Characterization of two novel highly active glycoside hydrolase family 53 endo-1,4-β-galactanases and their synergism with other carbohydrases in plant polysaccharide decomposition

被引：3

作者：

Zhang, Yurou ^{[1
,2
]}

Liu, Yun ^{[1
,2
]}

Zeng, Chunxiu ^{[1
,2
]}

Shu, Yingyu ^{[1
,2
]}

Wang, Xiaoru ^{[1
,2
]}

Liang, Shuangcheng ^{[1
,2
]}

Wang, Sidi ^{[3
]}

Zhan, Ruoting ^{[1
,2
]}

Wang, Kui ^{[1
,2
]}

机构：

[1] Guangzhou Univ Chinese Med, Sch Pharmaceut Sci, Res Ctr Chinese Herbal Resource Sci & Engn, Guangzhou 510006, Guangdong, Peoples R China

[2] Guangzhou Univ Chinese Med, Key Lab Chinese Med Resource Lingnan, Minist Educ Peoples Republ China, Guangzhou 510006, Guangdong, Peoples R China

[3] Guangzhou Univ Chinese Med, Coll Fundamental Med Sci, Guangzhou 510006, Guangdong, Peoples R China

来源：

INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES | 2023年 / 224卷

基金：

中国国家自然科学基金;

关键词：

GH53; enzymes; Endogalactanase; Galactan; Arabinogalactan; Synergy; ASPERGILLUS-ACULEATUS; THERMOSTABILITY; DEGRADATION; XYLANASE; ENZYMES; SITE; ENDO-BETA-1,4-GALACTANASE; PURIFICATION; PRETREATMENT; EXPRESSION;

D O I：

10.1016/j.ijbiomac.2022.10.154

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Two novel Bacillus glycoside hydrolase family 53 (GH53) endo-1,4-beta-galactanases (Bs936 and Bs4828) were identified. The recombinant Bs936 and Bs4828 displayed maximal activities at pH 5.5/55 degrees C and pH 6.5/65 degrees C, respectively. Stability analyses revealed that the enzymes were stable between pH 4.5-10, retaining over 80% activities after 12 h incubation at 25 degrees C. Moreover, Bs936 maintained about 75% activity after being treated at 45 degrees C for 2 h, while Bs4828 kept full activity after 4 h of incubation at 50 degrees C. Importantly, Bs936 and Bs4828 exhibited good activity towards beta-1,4-galactan, showing specific activities of 1859.46-and 3110.79 U/mg to-wards potato galactan, respectively. The corresponding Km and kcat/Km were 4.93 mg/mL and 296.35 mL/mg/s for Bs936, and 6.60 mg/mL and 707.12 mL/mg/s for Bs4828. Extensive synergy assays revealed that each endo-1,4-beta-galactanase acted synergistically with the other beta-galactosidase (Bs937 or Bs4829), endo-1,5-alpha-L-arabi-nosidase (BsAbn2) and an endoxylanase/endoglucanase+cellulase mixture (BaXynA + LsCel) towards the hy-drolysis of galactan, debranched arabinan/arabinan, and pretreated corn stover, respectively. The corresponding degrees of synergy were 1.28, 1.17/1.09, and 1.15 for Bs486 and 1.55, 1.04/1.08, and 1.11 for Bs4828, respectively. This study expands the repertoire of GH53 enzymes, and demonstrates the role of two highly active endo-1,4-beta-galactanases in polysaccharides hydrolysis.

引用

页码：653 / 666

页数：14

共 1 条

[1] Crystallographic analysis shows substrate binding at the -3 to +1 active-site subsites and at the surface of glycoside hydrolase family 11 endo-1,4-β-xylanases
Vandermarliere, Elien
Bourgois, Tine M.
Rombouts, Sigrid
Van Campenhout, Steven
Volckaert, Guido
Strelkov, Sergei V.
Delcour, Jan A.
Rabijns, Anja
Courtin, Christophe M.
BIOCHEMICAL JOURNAL, 2008, 410 : 71 - 79

← 1 →