L-Arginine is essential amino acid that is widely used in medicine, food addition, and other industries. The metabolic pathway engineering of Corynebacterium crenatum SYPA5-5 was performed to further enhance the flux of L-arginine biosynthesis. Comparing the enzymatic properties of acetyl-CoA synthase (ACS) from different bacterial sources, and heterologous expression in C. crenatum SYPA-ACS to effectively utilize acetic acid and increase L-arginine yield. Firstly, the four genes encoding ACS from different bacterial sources were cloned and expressed in Escherichia coli, and the highest activity of ApACS1 from Acetobacter pasteurianus ATCC 33445 was 784.59 U/mL, the optimum pH was 7.0 and temperature was 37 ℃. Compared with the other ACSs, ApACS1 has better pH and temperature stability. The acs1 from A. pasteurianus was then cloned into plasmid pXMJ19 and expressed in C. crenatum, which was effectively improving the acetyl-CoA content. The recombinant strain SYPA-ACS was cultured in 5 L fermenter for 96 h, L-arginine accumulation increased to 53.43 g/L with the productivity of 0.577 g/(L·h) in the recombinant SYPA-ACS, which was approximately 27.48% higher than that of SYPA5-5. The ACS from A. pasteurianus has been screened and overexpressed in C. crenatum, which could use acetic acid to improve the content of acetyl-CoA and achieve L-arginine accumulation. Thus, this strategy provides a safe and efficient synthesis of L-arginine and has important industrial application potential.
WU Jieqin
,
SHI Dian
,
XU Hua
,
ZHANG Xian
,
YANG Taowei
,
XU Meijuan
,
RAO Zhiming
. Enhancement of L-arginine yield by heterologously expressing acetyl-CoA synthetase in Corynebacterium crenatum[J]. Food and Fermentation Industries, 2022
, 48(21)
: 9
-16
.
DOI: 10.13995/j.cnki.11-1802/ts.030952
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