US 12,264,350 B2
Method for producing sulfated polysaccharide and method for producing PAPS
Kenta Fukunaga, Tokyo (JP); Aki Nishibara, Tokyo (JP); Ryosuke Kato, Tokyo (JP); Masahiro Kuratsu, Tokyo (JP); Mikiro Hayashi, Tokyo (JP); and Shin-ichi Hashimoto, Tokyo (JP)
Assigned to RENSSELAER POLYTECHNIC INSTITUTE, Troy, NY (US); OTSUKA PHARMACEUTICAL FACTORY, INC., Naruto (JP); and KIRIN BIOMATERIALS CO., LTD., Tokyo (JP)
Appl. No. 17/995,367
Filed by RENSSELAER POLYTECHNIC INSTITUTE, Troy, NY (US); OTSUKA PHARMACEUTICAL FACTORY, INC., Naruto (JP); and KIRIN BIOMATERIALS CO., LTD., Tokyo (JP)
PCT Filed Apr. 2, 2021, PCT No. PCT/JP2021/014337
§ 371(c)(1), (2) Date Oct. 3, 2022,
PCT Pub. No. WO2021/201282, PCT Pub. Date Oct. 7, 2021.
Claims priority of application No. PCT/JP2020/015388 (WO), filed on Apr. 3, 2020.
Prior Publication US 2023/0159969 A1, May 25, 2023
Int. Cl. C12P 19/04 (2006.01); C12N 9/12 (2006.01); C12P 19/26 (2006.01)
CPC C12P 19/26 (2013.01) [C12N 9/1205 (2013.01); C12N 9/1241 (2013.01); C12P 19/04 (2013.01); C12Y 207/01025 (2013.01); C12Y 207/07004 (2013.01)] 10 Claims
 
1. A method for producing a sulfated polysaccharide, the method comprising the following steps (1-1) and (1-2):
(1-1) preparing a transformant (a) of a bacterium of the genus Corynebacterium, comprising at least a gene encoding an adenosine 5′-triphosphate (ATP) sulfurylase and a gene encoding an adenosine 5′-phosphosulfate (APS) kinase which are introduced thereinto in an expressible manner and in which gene expression of an enzyme related to degradation of 3′-phosphoadenosine 5′-phosphosulfate (PAPS) is not attenuated, or a treated matter of the transformant (a), wherein the treated matter of transformant (a) is a matter in which the cell plasma membrane of the transformant (a) is substance permeable, and wherein the transformant (a) or the treated matter thereof is capable of producing PAPS; and
(1-2) conducting a reaction for producing PAPS by using a reaction solution containing ATP or an ATP source, a sulfate ion source, and the transformant (a) or the treated matter of transformant (a), wherein the enzyme related to degradation of PAPS is not heat inactivated; and
wherein the method further comprises conducting sulfation by a transformant comprising a gene encoding sulfotransferase, which is introduced thereinto in an expressible manner, or a treated matter or extract of the transformant comprising a gene encoding sulfotransferase.