US 11,739,357 B2
Recombinant microorganism for producing citicoline and method for producing citicoline
Junjun Jiang, Jiangsu (CN); Junying Fan, Jiangsu (CN); Feng Tian, Jiangsu (CN); Xintong Wang, Jiangsu (CN); Kailin Zhang, Jiangsu (CN); and Zhihao Hu, Castro Valley, CA (US)
Assigned to SUZHOU BIOSYNTHETICA CO., LTD, Suzhou (CN)
Filed by SUZHOU BIOSYNTHETICA CO., LTD, Jiangsu (CN)
Filed on Jan. 7, 2020, as Appl. No. 16/736,781.
Application 16/736,781 is a continuation of application No. PCT/CN2018/075238, filed on Feb. 5, 2018.
Claims priority of application No. 201710549120.0 (CN), filed on Jul. 7, 2017.
Prior Publication US 2020/0140910 A1, May 7, 2020
Int. Cl. C12P 19/30 (2006.01); C12N 1/20 (2006.01); C12N 9/12 (2006.01); C12N 15/70 (2006.01)
CPC C12P 19/305 (2013.01) [C12N 1/20 (2013.01); C12N 9/1205 (2013.01); C12N 9/1241 (2013.01); C12N 15/70 (2013.01); C12Y 207/01032 (2013.01); C12Y 207/07015 (2013.01)] 3 Claims
 
1. A genetically modified Escherichia coli for producing citicoline comprising the following modifications:
a. enzymes involved in reuse of citicoline, or choline, or phosphocholine being disrupted, wherein the enzymes comprise cytidine-5′-diphosphoinositol hydrolase, cytidine-5′-diphosphate-diacylglycerol pyrophosphatase, choline dehydrogenase, alkaline phosphatase or acid phosphatase;
b. choline kinase for catalyzing choline chloride to generate phosphocholine being overexpressed, wherein the choline kinase comprises CKI1 or EKI1 obtained from Saccharomyces cerevisiae, or LicC obtained from Streptococcus;
c. cytidylyltransferase for catalyzing phosphocholine to generate citicoline being overexpressed, wherein the cytidylyltransferase comprises PCT1, CDS1, or ECT1 obtained from Saccharomyces cerevisiae; or CdsA, IspD, MocA, KdsB, or Cca obtained from Escherichia coli; or LicC obtained from Streptococcus; or CD36_40620 obtained from Candida dubliniensis;
d. choline transporter protein, for transporting choline chloride into a cell being overexpressed, wherein the choline transporter protein comprises BetT obtained from Escherichia coli; and
e. uridine-5′-monophosphate phosphorylase for degrading uridine-5′-monophosphate into uridine is disrupted; and the uridine-5′-monophosphate phosphorylase comprises UmpH, UmpG, PhoA, AphA, or YjjG.