Production Optimization of an Active β-Galactosidase of Bifidobacterium animalis in Heterologous Expression Systems
Joint Authors
Xu, Xinxin
Fan, Xiaohu
Fan, Chao
Qin, Xing
Liu, Bo
Nie, Chunming
Sun, Ning
Yao, Qingzhi
Zhang, Yuhong
Zhang, Wei
Source
Issue
Vol. 2019, Issue 2019 (31 Dec. 2019), pp.1-10, 10 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2019-02-20
Country of Publication
Egypt
No. of Pages
10
Main Subjects
Abstract EN
β-Galactosidase (E.C.3.2.1.23) catalyzes the hydrolysis of lactose into glucose and galactose and the synthesis of galacto-oligosaccharides as well.
The β-galactosidases from bacteria, especially lactobacilli, and yeast have neutral pH and are much more likely to be developed as food additives.
However, the challenges of cumbersome purification, product toxicity, and low yield in protein production have limited the commercialization of many excellent candidates.
In this study, we identified a β-galactosidase gene (bg42-106) in Bifidobacterium animalis ACCC05790 and expressed the gene product in Escherichia coli BL21(DE3) and Pichia pastoris GS115, respectively.
The recombinant bG42-106 purified from E.
coli cells was found to be optimally active at pH 6.0 and 60°C and had excellent stability over a wide pH range (5.0–8.0) and at high temperature (60°C).
The specific activity of bG42-106 reached up to 2351 U/mg under optimal conditions.
The galacto-oligosaccharide yield was 24.45 g/L after incubation with bG42-106 at 60°C for 2 h.
When recombinant bG42-106 was expressed in Pichia pastoris GS115, it was found in the culture medium but only at a concentration of 1.73 U/ml.
To increase its production, three strategies were employed, including codon optimization, disulfide formation, and fusion with a Cherry tag, with Cherry-tag fusion being most effective.
The culture medium of P.
pastoris that expressed Cherry-tagged bG42-106 contained 24.4 U/mL of β-galactosidase activity, which is 14-fold greater than that produced by culture of P.
pastoris harboring wild-type bG42-106.
American Psychological Association (APA)
Xu, Xinxin& Fan, Xiaohu& Fan, Chao& Qin, Xing& Liu, Bo& Nie, Chunming…[et al.]. 2019. Production Optimization of an Active β-Galactosidase of Bifidobacterium animalis in Heterologous Expression Systems. BioMed Research International،Vol. 2019, no. 2019, pp.1-10.
https://search.emarefa.net/detail/BIM-1127652
Modern Language Association (MLA)
Xu, Xinxin…[et al.]. Production Optimization of an Active β-Galactosidase of Bifidobacterium animalis in Heterologous Expression Systems. BioMed Research International No. 2019 (2019), pp.1-10.
https://search.emarefa.net/detail/BIM-1127652
American Medical Association (AMA)
Xu, Xinxin& Fan, Xiaohu& Fan, Chao& Qin, Xing& Liu, Bo& Nie, Chunming…[et al.]. Production Optimization of an Active β-Galactosidase of Bifidobacterium animalis in Heterologous Expression Systems. BioMed Research International. 2019. Vol. 2019, no. 2019, pp.1-10.
https://search.emarefa.net/detail/BIM-1127652
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1127652