High yields of galactooligosaccharides may be obtained by enzymatic hydrolysis of potato pulp using endo-β-1,4-galactanase (EC 3.2.1.89). In present study, the gene aghA encoding endo-β-1,4-galactanase from Aspergillus niger CICIM F0510 was thus cloned and expressed in Pichia pastoris, which generated the recombinant strain GS115 (pPIC-aghA). In shaking flask experiments, the activity of recombinant enzyme AghA was 130 U/mL. The optimal pH and temperature of AghA were 4.5 and 45 ℃, respectively, and it was stable at pH 4.0-6.0 or at 30-50 ℃. Most of chemical ions and EDTA had no significant effects on the activity of AghA, while it was strongly inhibited by Fe3+ and nearly completely lost in the presence of Hg2+. The Vmax and Km of AghA towards galactan (potato) were determined to be 400 mg/(mL·min) and 0.08 mg/mL, respectively. Besides, potato pulp was hydrolyzed by AghA into galactobiose, galactotrioses and a small amount of galactotetraose. The results obtained paved the way for large scale production of galactanase AghA and its applications in preparing galactooligosaccharides from potato pulp.
CAI Ke
,
WANG Taikang
,
WANG Jun
,
DONG Zixing
,
TIAN Kangming
,
JIN Peng
,
LIU Xiaoguang
,
WANG Zhengxiang
. Molecular cloning and biochemical characterization of endo-beta-1,4-galactanase AghA from Aspergillus niger[J]. Food and Fermentation Industries, 2019
, 45(1)
: 29
-35
.
DOI: 10.13995/j.cnki.11-1802/ts.017756
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