Enhancement of pipecolic
acid production by the expression of multiple lysine cyclodeaminase in
the Escherichia coli whole-cell system
Yeong-Hoon Han, Tae-Rim Choi,
Ye-Lim Park, Jun Young Park, Hun-Suk Song, Hyun-Joong Kim, Sun Mi Lee,
Sol Lee Park, Hye Soo Lee, Shashi Kant Bhatia, Ranjit Gurav,
Yung-Hun Yang*
Department of Biological
Engineering, College of Engineering, Konkuk University, 1 Hwayang-dong,
Gwangjin-gu, Seoul 05029, Republic of Korea
*Corresponding author (E-mail:seokor@konkuk.ac.kr )
Running title: Reinforced-LCD improves pipecolic acid production
ABSTRACT
Pipecolic acid, a non-proteinogenic amino acid, is a metabolite in
lysine metabolism and a key chiral precursor in local anesthesia and
macrolide antibiotics. To replace the environmentally unfriendly
chemical production or preparation procedure of pipecolic acid, many
biological synthetic routes have been studied for a long time. Among
them, synthesis by lysine cyclodeaminase (LCD), encoded by pipA ,
has several advantages, including stability of enzyme activity and
NAD+ self-regeneration. Thus, we selected this enzyme
for pipecolic acid biosynthesis in a whole-cell bioconversion. To
construct a robust pipecolic acid production system, we investigated
important conditions including expression vector, strain, culture
conditions, and other reaction parameters. The most important factors
were introduction of multiple pipA genes into the whole-cell
system and control of agitation. As a result, we produced 724 mM
pipecolic acid (72.4% conversion), and the productivity was 0.78 g/L/h
from 1 M l-lysine after 5 days. This is the highest production
reported to date.
Keywords : pipecolic acid (l-PA), pharmaceutical
precursor, whole-cell bioconversion, lysine cyclodeaminse (LCD),
reinforced enzyme