Affinity induced immobilization of adenylate cyclase from the crude cell lysate for ATP conversion

Zhu, Qianqian and Zhuang, Wei and Niu, Huanqing and Ge, Lei and Hernandez, Byron Villacorta and Wu, Jinglan and Wang, Kai and Liu, Dong and Chen, Yong and Zhu, Chenjie and Ying, Hanjie (2018) Affinity induced immobilization of adenylate cyclase from the crude cell lysate for ATP conversion. Colloids and Surfaces B: Biointerfaces, 164. pp. 155-164. ISSN 0927-7765

Abstract

The development of an orientation immobilization technique via affinity between polyhistidine tags and metal ions aims at maintaining biocatalytic activity of the enzymes. In this work, to tackle the issue of the immobilization of adenylate cyclase (AC), a simple and effective approach of synthesizing iminodiacetic acid (IDA)-Ni2+ particles was applied for simultaneously purifying and immobilizing his-tagged AC. We chose agarose particles as carriers, and then decorated them with IDA, leading to the formation of a coordination combination of Ni2+. The porous carriers with a large pore size of 50 nm and a specific surface area of 45.8 m2/g exhibited favorable enzymatic activity and loading capacity. The optimal pH of the immobilized enzyme increased from 8.0 to 9.0 and the optimal temperature increased from 30 °C to 35 °C, compared to the free AC. Moreover, the immobilized AC retained a residual activity of approximately 80% after storing it at 25 °C for 48 h, whereas only 40% of the activity was left in the free AC at the same conditions. Maximum yield of cyclic adenosine-3′, 5′- monophosphate (cAMP) reached up to the summit of the reaction. The immobilized AC by affinity adsorption will provide a promising route for the industrial production of cAMP.


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Item Type: Article (Commonwealth Reporting Category C)
Refereed: Yes
Item Status: Live Archive
Additional Information: Access to published version in accordance with the copyright policy of the publisher.
Faculty / Department / School: Current - Faculty of Health, Engineering and Sciences - School of Mechanical and Electrical Engineering
Date Deposited: 03 Apr 2018 05:51
Last Modified: 03 Apr 2018 05:51
Uncontrolled Keywords: adenylate cyclase; cyclic adenosine-3′,5′-monophosphate; iminodiacetic acid; his-tagged; enzyme immobilization; nickel ions
Fields of Research : 03 Chemical Sciences > 0306 Physical Chemistry (incl. Structural) > 030603 Colloid and Surface Chemistry
03 Chemical Sciences > 0306 Physical Chemistry (incl. Structural) > 030601 Catalysis and Mechanisms of Reactions
Socio-Economic Objective: B Economic Development > 86 Manufacturing > 8606 Industrial Chemicals and Related Products > 860699 Industrial Chemicals and Related Products not elsewhere classified
Identification Number or DOI: 10.1016/j.colsurfb.2018.01.033
URI: http://eprints.usq.edu.au/id/eprint/33910

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