Good hydrolysis activity on raffinose family oligosaccharides by a novel α-galactosidase from Tremella aurantialba

Xueran Geng; Dongxue Yang; Qiaoyi Zhang; Mingchang Chang; Lijing Xu; Yanfen Cheng; Hexiang Wang; Junlong Meng

doi:10.1016/j.ijbiomac.2019.10.136

Good hydrolysis activity on raffinose family oligosaccharides by a novel α-galactosidase from Tremella aurantialba

Int J Biol Macromol. 2020 May 1:150:1249-1257. doi: 10.1016/j.ijbiomac.2019.10.136. Epub 2019 Nov 15.

Authors

Xueran Geng¹, Dongxue Yang², Qiaoyi Zhang³, Mingchang Chang⁴, Lijing Xu⁴, Yanfen Cheng⁴, Hexiang Wang⁵, Junlong Meng⁶

Affiliations

¹ College of Food Science and Engineering, Shanxi Agricultural University, Taigu, Shanxi 030801, China; Collaborative Innovation Center of Advancing Quality and Efficiency of Loess Plateau Edible Fungi, Taigu, Shanxi 030801, China; State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University, Beijing 100193, China.
² State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University, Beijing 100193, China.
³ Orient Science & Technology College of Hunan Agricultural University, China.
⁴ College of Food Science and Engineering, Shanxi Agricultural University, Taigu, Shanxi 030801, China; Collaborative Innovation Center of Advancing Quality and Efficiency of Loess Plateau Edible Fungi, Taigu, Shanxi 030801, China.
⁵ State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University, Beijing 100193, China. Electronic address: hxwang@cau.edu.cn.
⁶ College of Food Science and Engineering, Shanxi Agricultural University, Taigu, Shanxi 030801, China; Collaborative Innovation Center of Advancing Quality and Efficiency of Loess Plateau Edible Fungi, Taigu, Shanxi 030801, China. Electronic address: mengjunlongseth@126.com.

PMID: 31739012
DOI: 10.1016/j.ijbiomac.2019.10.136

Abstract

An α-galactosidase designated as TAG was purified from the dried fruit bodies of Tremella aurantialba with 182.5-fold purification. The purification procedure involved ion exchange chromatography on Q-sepharose, DEAE-Cellulose, and Mono Q and gel filtration by FPLC on Superdex 75. The purified α-galactosidase was a monomeric protein with a molecular mass of 88 kDa. The optimal pH of TAG was 5.0 and more than 60% of the original enzyme activity remained at pH 2.0 and 3.0. Its optimal temperature was 54 °C with good thermo-stability, 30.8% of the original activity was retained after exposure to a temperature of 70 °C for 1 h. The metal ions Hg²⁺, Cu²⁺, Fe³⁺ and Mg²⁺ strongly inhibited the enzyme activity. The enzyme activity was found to be inhibited by N-bromosuccinimide indicating that tryptophan was essential to the catalytic activity of α-galactosidase. The enzyme completely hydrolysed stachyose and partially hydrolysed raffinose to galactose at 50 °C within 6 h as detected by thin layer chromatography and the dinitrosalicylic acid method and the content of reducing sugar reached 4.36 mg/mL.

Keywords: Mushroom; Oligosaccharides; Tremella aurantialba; α-galactosidase.

MeSH terms

Basidiomycota / enzymology*
Enzyme Stability
Fungal Proteins* / chemistry
Fungal Proteins* / isolation & purification
Hot Temperature
Hydrolysis
Metals / chemistry
Oligosaccharides / chemistry*
Raffinose / chemistry*
alpha-Galactosidase* / chemistry
alpha-Galactosidase* / isolation & purification

Substances

Fungal Proteins
Metals
Oligosaccharides
alpha-Galactosidase
Raffinose

Supplementary concepts

Naematelia aurantialba