TY - JOUR
T1 - Efficient production of 5-aminovalerate from L-lysine by engineered Escherichia coli whole-cell biocatalysts
AU - Wang, Xin
AU - Cai, Peipei
AU - Chen, Kequan
AU - Ouyang, Pingkai
N1 - Publisher Copyright:
© 2016 Elsevier B.V.
PY - 2016/12/1
Y1 - 2016/12/1
N2 - In this study, we developed a whole-cell biocatalysis process for high-level conversion of L-lysine into 5-aminovalerate. To obtain the highly efficient whole-cell biocatalyst, five expression plasmids were constructed to optimize the expression of 5-aminovaleramide amidohydrolase and L-lysine 2-monooxygenase in Escherichia coli. The engineered strain BL-22A-RB-YB harboring plasmid pET22b-davA, pRSFDuet-davB and pACYCDuet-davB was correspondingly obtained. Subsequently, the effects of induction conditions, reaction temperature, metal ion additives, and cell permeability on the whole-cell biocatalyst system were evaluated to improve biocatalytic efficiency. Under optimized reaction conditions, 95.3 g/L 5-aminovalerate was synthesized from 120 g/L L-lysine with a yield of 99.1%, and 103.1 g/L 5-aminovalerate was produced from 150 g/L L-lysine with a molar yield of 85.7%. The 5-aminovalerate production was then further improved using a L-lysine fed-batch strategy, and a hyper 5-aminovalerate production of 240.7 g/L was achieved within 28 h with a yield of 86.8%. The whole-cell biocatalytic system described here demonstrated an environmentally friendly strategy for industrial production of 5-aminovalerate.
AB - In this study, we developed a whole-cell biocatalysis process for high-level conversion of L-lysine into 5-aminovalerate. To obtain the highly efficient whole-cell biocatalyst, five expression plasmids were constructed to optimize the expression of 5-aminovaleramide amidohydrolase and L-lysine 2-monooxygenase in Escherichia coli. The engineered strain BL-22A-RB-YB harboring plasmid pET22b-davA, pRSFDuet-davB and pACYCDuet-davB was correspondingly obtained. Subsequently, the effects of induction conditions, reaction temperature, metal ion additives, and cell permeability on the whole-cell biocatalyst system were evaluated to improve biocatalytic efficiency. Under optimized reaction conditions, 95.3 g/L 5-aminovalerate was synthesized from 120 g/L L-lysine with a yield of 99.1%, and 103.1 g/L 5-aminovalerate was produced from 150 g/L L-lysine with a molar yield of 85.7%. The 5-aminovalerate production was then further improved using a L-lysine fed-batch strategy, and a hyper 5-aminovalerate production of 240.7 g/L was achieved within 28 h with a yield of 86.8%. The whole-cell biocatalytic system described here demonstrated an environmentally friendly strategy for industrial production of 5-aminovalerate.
KW - 5-Aminovaleramide amidohydrolase
KW - 5-Aminovalerate
KW - L-Lysine
KW - L-Lysine 2-monooxygenase
KW - Whole-cell bioconversion
UR - http://www.scopus.com/inward/record.url?scp=84992028817&partnerID=8YFLogxK
U2 - 10.1016/j.molcatb.2016.10.008
DO - 10.1016/j.molcatb.2016.10.008
M3 - 文章
AN - SCOPUS:84992028817
SN - 1381-1177
VL - 134
SP - 115
EP - 121
JO - Journal of Molecular Catalysis - B Enzymatic
JF - Journal of Molecular Catalysis - B Enzymatic
ER -