[Technical Field]
The present disclosure relates to a putrescine-producing microorganism and a
method for producing putrescine by using same.
5 [Background Art]
Putrescine, which is a raw material of nylon, is produced mainly by a chemical
method using petroleum compounds as raw materials. Specifically, putrescine is produced
by adding hydrogen cyanide to acrylonitrile to prepare succinonitrile, followed by
hydrogenation. Such a chemical process has efficiency and economic feasibility, but has
10 the disadvantage of being environmentally unfriendly. Hence, due to the strengthening of
environmental regulations and the above, there is a need for the production of alternative
substances through bio-based pathways.
In relation to this, methods for producing high concentrations of putrescine by
transformation of E. coli and microorganisms of the genus Corynebacterium are disclosed
15 (Morris et al., J Biol. Chem. 241: 13, 3129-3135, 1996; WO 06/005603; WO 09/125924;
Qian ZD et al., Biotechnol. Bioeng. 104: 4, 651-662, 2009; Schneider et al., Appl. Microbiol.
Biotechnol. 88: 4, 859-868, 2010; and Schneider et al., Appl. Microbiol. Biotechnol. 91: 17-
30, 2011). Additionally, various methods for producing putrescine using microorganisms
are known (US 13/992242, US 14/372000, US 14/373265, EP 2236613 B1, and US
20 8497098 B2).
Among the proteins associated with the putrescine biosynthesis pathway, argJ,
bifunctional ornithine acetyltransferase/N-acetylglutamate synthase, is an enzyme that can
perform a conversion reaction of two substances, acetyl-CoA and N-acetylornithine and has
functions of both ornithine acetyltransferase (L-glutamate N-acetyltransferase) and N25 acetylglutamate synthase and has both functions of an ornithine acetyltransferase and an Nacetylglutamate synthase. The enzyme argJ can reduce byproducts and lessen the burden
of putrescine production since one enzyme intermediates two enzymatic reactions.
However, the activity of argJ may be inhibited by intermediate metabolites, such as
ornithine (Sakanyan V, Petrosyan P, Lecocq M, Boyen A, Legrain C, Demarez M, Hallet J,
30 Glansdorff N: Genes and enzymes of the acetyl cycle of arginine biosynthesis in
Corynebacterium glutamicum: enzyme evolution in the early steps of the arginine pathway.
Microbiology 1996, 142:99-108), resulting in lowered biosynthesis efficiency of putrescine.
N-acetyltransferase present in Corynebacterium glutamicum has N-acetyl-L-
3
glutamate producing ability in the presence of acetyl-CoA and glutamate. However, it has
been reported that the N-acetyltransferase has at least 9.43 times higher specific activity
than argJ (A new type of N-acetylglutamate synthase is involved in the first step of arginine
biosynthesis in Corynebacterium glutamicum. "BMC genomics 2013(14) p 713).

We Claim:
1. A microorganism of the genus Corynebacterium having putrescine producing ability,
into which activity of an N-acetyltransferase derived from a strain of the genus
5 Corynebacterium and activity of an acetylornithine deacetylase (argE) derived from E.
coli are introduced.
2. The microorganism of the genus Corynebacterium of claim 1, wherein the Nacetyltransferase derived from the strain of the genus Corynebacterium comprises the
10 amino acid sequence of SEQ ID NO: 1 or an amino acid sequence having sequence
identity of at least 90% thereto.
3. The microorganism of the genus Corynebacterium of claim 1, wherein the
acetylornithine deacetylase (argE) derived from E. coli comprises the amino acid
15 sequence of SEQ ID NO: 3 or an amino acid sequence having sequence identity of at
least 90% thereto.
4. The microorganism of the genus Corynebacterium of claim 1, wherein the
microorganism has weakened activity of bifunctional ornithine acetyltransferase/N20 acetylglutamate synthase (argJ) derived from a strain of the genus Corynebacterium.
5. The microorganism of the genus Corynebacterium of claim 1, wherein the
microorganism is Corynebacterium glutamicum.
25 6. A method for producing putrescine, the method comprising culturing, in a medium, a
microorganism of the genus Corynebacterium having putrescine producing ability,
into which activity of an N-acetyltransferase derived from a strain of the genus
Corynebacterium and activity of an acetylornithine deacetylase (argE) derived from E.
coli are introduced.
30
7. The method of claim 6, further comprising recovering putrescine from the cultured
medium or microorganism.
8. The method of claim 6, wherein the N-acetyltransferase derived from the strain of the
38
genus Corynebacterium has the amino acid sequence of SEQ ID NO: 1.
9. The method of claim 6, wherein the acetylornithine deacetylase derived from E. coli
has the amino acid sequence of SEQ ID NO: 3.
5
10. A composition for producing putrescine, the composition comprising a microorganism
of the genus Corynebacterium having putrescine producing ability, into which activity
of an N-acetyltransferase derived from a strain of the genus Corynebacterium and
activity of an acetylornithine deacetylase (argE) derived from E. coli are introduced.
10
11. The composition of claim 10, wherein the N-acetyltransferase derived from the strain
of the genus Corynebacterium comprises the amino acid sequence of SEQ ID NO: 1
or an amino acid sequence having sequence identity of at least 90% thereto.
15 12. The composition of claim 10, wherein the acetylornithine deacetylase derived from E.
coli comprises the amino acid sequence of SEQ ID NO: 3 or an amino acid sequence
having sequence identity of at least 90% thereto.
13. Use of a microorganism of the genus Corynebacterium having putrescine producing
20 ability for the production of putrescine, wherein activity of an N-acetyltransferase
derived from a strain of the genus Corynebacterium and activity of an acetylornithine
deacetylase (argE) derived from E. coli are introduced into the microorganism.