TY - JOUR
T1 - Migration of glutamate decarboxylase by cold treatment on whole-cell biocatalyst triggered activity for 4-aminobutyric acid production in engineering Escherichia coli
AU - Xue, Chengfeng
AU - Yi, Ying Chen
AU - Ng, I. Son
N1 - Funding Information:
The authors are grateful for the financial support for this study provided by the Ministry of Science and Technology of Taiwan (MOST 110-2221-E-006-030-MY3 and MOST 108-2221-E-006-004-MY3). The authors gratefully thank the use of Qrbitrap LC-MS-MS [MS004000] belonging to the Core Facility Center of National Cheng Kung University.
Funding Information:
The authors are grateful for the financial support for this study provided by the Ministry of Science and Technology of Taiwan ( MOST 110-2221-E-006-030-MY3 and MOST 108-2221-E-006-004-MY3 ). The authors gratefully thank the use of Qrbitrap LC-MS-MS [MS004000] belonging to the Core Facility Center of National Cheng Kung University.
Publisher Copyright:
© 2021 Elsevier B.V.
PY - 2021/11/1
Y1 - 2021/11/1
N2 - Glutamate decarboxylase B (GadB) from Escherichia coli, an intrinsic pyridoxal 5′-phosphate (PLP)-dependent enzyme has been employed for 4-aminobutyric acid (GABA) biosynthesis, which involves the glutamate import and GABA export via a transporter located in the inner membrane as rate determined step of whole-cell (WC) biotransformation. Herein, GadB was cloned and overexpressed in E. coli under a constitutive promoter in a high copy number plasmid, and 46.9 g/L GABA was produced. It was observed that GadB migrated to the periplasm when the WC were subjected to −20 °C cold treatment for 24 h prior to the biotransformation. Kinetic studies indicated that the enzymatic turnover rate of WC increased 2-fold after cold treatment, which was correlated with the migration rate of GadB, and up to 88.6% of GadB. The export or possible migration of GadB mitigated the rate-limiting step of WC biotransformation, and a 100% conversion of substrate to GABA was obtained. Finally, we launched a promising strategy for GABA production of 850 g/L from cost-effective monosodium glutamate (MSG) by using WC biocatalysts with 10-times recycling.
AB - Glutamate decarboxylase B (GadB) from Escherichia coli, an intrinsic pyridoxal 5′-phosphate (PLP)-dependent enzyme has been employed for 4-aminobutyric acid (GABA) biosynthesis, which involves the glutamate import and GABA export via a transporter located in the inner membrane as rate determined step of whole-cell (WC) biotransformation. Herein, GadB was cloned and overexpressed in E. coli under a constitutive promoter in a high copy number plasmid, and 46.9 g/L GABA was produced. It was observed that GadB migrated to the periplasm when the WC were subjected to −20 °C cold treatment for 24 h prior to the biotransformation. Kinetic studies indicated that the enzymatic turnover rate of WC increased 2-fold after cold treatment, which was correlated with the migration rate of GadB, and up to 88.6% of GadB. The export or possible migration of GadB mitigated the rate-limiting step of WC biotransformation, and a 100% conversion of substrate to GABA was obtained. Finally, we launched a promising strategy for GABA production of 850 g/L from cost-effective monosodium glutamate (MSG) by using WC biocatalysts with 10-times recycling.
UR - https://www.scopus.com/pages/publications/85114173493
UR - https://www.scopus.com/pages/publications/85114173493#tab=citedBy
U2 - 10.1016/j.ijbiomac.2021.08.166
DO - 10.1016/j.ijbiomac.2021.08.166
M3 - Article
C2 - 34480902
AN - SCOPUS:85114173493
SN - 0141-8130
VL - 190
SP - 113
EP - 119
JO - International Journal of Biological Macromolecules
JF - International Journal of Biological Macromolecules
ER -