TY - JOUR
T1 - Efficient production of cyclic adenosine monophosphate from adenosine triphosphate by the N-terminal half of adenylate cyclase from Escherichia coli
AU - Ma, Chen
AU - Wang, Jing
AU - Wang, Xuelin
AU - Mai, Dandan
AU - Jin, Yuqi
AU - Chen, Kequan
AU - Wang, Xin
AU - Ouyang, Pingkai
N1 - Publisher Copyright:
© 2020 Elsevier B.V.
PY - 2020/8
Y1 - 2020/8
N2 - In this study, we aimed at developing an efficient biocatalytic process for bio-production of cyclic adenosine monophosphate (cAMP) from adenosine triphosphate (ATP). First, adenylate cyclase from Escherichia coli MG1655 (EAC) and Bordetella Pertussis (BAC) were expressed in E. coli BL21 (DE3) and comparatively analyzed for their activities. As a result, EAC from E. coli MG1655 exhibited a higher activity. However, amount of EAC were obtained in an insoluble form. Therefore, we expressed the first 446 amino acids of EAC (EAC446) to avoid the inclusion body. The effects of induction temperature, incubation time, and incubation pH were further evaluated to improve the expression of EAC446. Subsequently, the reaction process for the production of cAMP with ATP as a starting material was investigated. As none of cAMP was detected in the whole-cell based biocatalytic process, the reaction catalyzed by the crude enzyme was determined for cAMP production. What's more, the reaction temperature, reaction pH, metal ion additives and substrate concentration was optimized, and the maximum cAMP production of 18.45 g·L−1 was achieved with a yield of 95.4% after bioconversion of 6 h.
AB - In this study, we aimed at developing an efficient biocatalytic process for bio-production of cyclic adenosine monophosphate (cAMP) from adenosine triphosphate (ATP). First, adenylate cyclase from Escherichia coli MG1655 (EAC) and Bordetella Pertussis (BAC) were expressed in E. coli BL21 (DE3) and comparatively analyzed for their activities. As a result, EAC from E. coli MG1655 exhibited a higher activity. However, amount of EAC were obtained in an insoluble form. Therefore, we expressed the first 446 amino acids of EAC (EAC446) to avoid the inclusion body. The effects of induction temperature, incubation time, and incubation pH were further evaluated to improve the expression of EAC446. Subsequently, the reaction process for the production of cAMP with ATP as a starting material was investigated. As none of cAMP was detected in the whole-cell based biocatalytic process, the reaction catalyzed by the crude enzyme was determined for cAMP production. What's more, the reaction temperature, reaction pH, metal ion additives and substrate concentration was optimized, and the maximum cAMP production of 18.45 g·L−1 was achieved with a yield of 95.4% after bioconversion of 6 h.
KW - Adenosine triphosphate (ATP)
KW - Adenylate cyclase
KW - Bioconversion
KW - Cyclic adenosine monophosphate (cAMP)
UR - http://www.scopus.com/inward/record.url?scp=85083061683&partnerID=8YFLogxK
U2 - 10.1016/j.cjche.2020.01.003
DO - 10.1016/j.cjche.2020.01.003
M3 - 文章
AN - SCOPUS:85083061683
SN - 1004-9541
VL - 28
SP - 2167
EP - 2172
JO - Chinese Journal of Chemical Engineering
JF - Chinese Journal of Chemical Engineering
IS - 8
ER -