International Journal of Applied Microbiology and Biotechnology Research
ISSN: 2053-1818
Vol. 5(1), pp. 1-11, January 2017



Surface display of human arginase1 through engineered antigen 43 in Escherichia coli

Rongxing Tang1, Zhen Zhang1, Lu Bian2, Fei Wang1,Qiuting Gong1, Jinhui Xue1 and Lixin Ma1*

1Hubei Collaborative Innovation Center for Green Transformation of Bio-resources, Hubei Key Laboratory of Industrial Biotechnology, College of Life Sciences, Hubei University, Wuhan, People’s Republic of China.
2Beihai Blood Center, Guang Xi, People’s Republic of China.

*To whom correspondence should be addressed. E-mail: malixing@hubu.edu.cn.

Received 19 November, 2016; Received in revised form 13 December, 2016; Accepted 15 December, 2016.

Abstract


Keywords:
Antigen 43, Human Arginase1, Charged polypeptides, Surface display.


Escherichia coli auto-transporter protein antigen 43 (Ag43) was one of the superfamily of auto-transporters (AT) which was considered to be a potentially useful anchor for bacteria surface display because of its relative simplicity and high copy numbers. Here, the system used four charged polypeptides [6×Lys, 6×Glu and 6×Asp and 2×(Lys/Lys/Arg)] was engineered to replace its native signal peptide for the surface display of human Arginase1 (ARG1) in E. coli. The results obtained indicate that charged polypeptides 6×Lys, 6×Glu and 2×(Lys/Lys/Arg) could significantly enhance the display efficiency of human ARG1. Among these tags, the 6×Lys polypeptide had the highest display efficiency (37.73%) which was about 7.6-fold of the original Ag43 system (4.91%). The whole cell enzyme activity test also confirmed that optimized system (K6-ARG1-Ag43) obtained the highest activity of 15.21 U/mL(OD600=1.0), which was about 22-fold of the original Ag 43 system (0.69 U/mL, OD600=1.0). Optimized system (K6-ARG1-Ag43) also possessed high stability, after 10 repeated reaction cycles, the whole cell enzyme activity remained more than 35% of its initial activity. Furthermore, almost 100% of L-arginine (200 g/L) transferred into L-ornithine (L-Orn) in 16 h under the optimum condition in the batch conversion test. The engineered Ag43 display system expanded the Ag43 surface display system and provided alternative methods for display of human Arginase1 in E. coli.

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