Enhanced vanillin production from recombinant E. coli using NTG mutagenesis and adsorbent resin.
Vanillin production was tested with different concentrations of added ferulic acid in E. coli harboring plasmid pTAHEF containing fcs (feruloyl-CoA synthase) and ech (enoyl-CoA hydratase/aldolase) genes cloned from Amycolatopsis sp. strain HR104. The maximum production of vanillin from E. coli DH5alpha harboring pTAHEF was found to be 1.0 g/L at 2.0 g/L of ferulic acid for 48 h of culture. To improve the vanillin production by reducing its toxicity, two approaches were followed: (1) generation of vanillin-resistant mutant of NTG-VR1 through NTG mutagenesis and (2) removal of toxic vanillin from the medium by XAD-2 resin absorption. The vanillin production of NTG-VR1 increased to three times at 5 g/L of ferulic acid when compared with its wild-type strain. When 50% (w/v) of XAD-2 resin was employed in culture with 10 g/L of ferulic acid, the vanillin production of NTG-VR1 was 2.9 g/L, which was 2-fold higher than that obtained with no use of the resin.
Yoon
Sang-Hwal
SH
Division of Applied Life Science (BK21) and Forest Science, EB-NCRC and PMBBRC, Gyeongsang National University, Jinju 660-701, Korea.
Lee
Eun-Gyeong
EG
Das
Amitabha
A
Lee
Sook-Hee
SH
Li
Cui
C
Ryu
Hee-Kyoung
HK
Choi
Myung-Suk
MS
Seo
Weon-Taek
WT
Kim
Seon-Won
SW
D000042:Absorption; D000180:Acrylic Resins / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D024861:Genetic Enhancement / Q000379:methods*; D008769:Methylnitronitrosoguanidine / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D011995:Recombination, Genetic / Q000502:physiology; D014462:Ultrafiltration / Q000379:methods*
D000180:Acrylic Resins; D008769:Methylnitronitrosoguanidine