	US 12,356,911 B2
	Group of UDP-glycosyltransferase for catalyzing carbohydrate chain elongation and application thereof
Zhihua Zhou, Shanghai (CN); Wei Wei, Shanghai (CN); Xing Yan, Shanghai (CN); Chengshuai Yang, Shanghai (CN); Chaojing Li, Shanghai (CN); Yongjun Wei, Shanghai (CN); and Pingping Wang, Shanghai (CN)
Assigned to GSYNBIOT (SHANGHAI) CO., LTD, Shanghai (CN)
Filed by GSYNBIOT (SHANGHAI) CO., LTD, Shanghai (CN)
Filed on Oct. 28, 2022, as Appl. No. 17/976,715.
Application 17/976,715 is a division of application No. 16/614,944, granted, now 11,542,484, previously published as PCT/CN2018/087678, filed on May 21, 2018.
Claims priority of application No. CN201710359069.7 (CN), filed on May 19, 2017.
Prior Publication US 2023/0203458 A1, Jun. 29, 2023
Int. Cl. A01H 1/00 (2006.01); A01H 5/06 (2018.01); C12N 9/10 (2006.01); C12P 19/18 (2006.01)

CPC A01H 5/06 (2013.01) [A01H 1/00 (2013.01); C12N 9/1048 (2013.01); C12P 19/18 (2013.01)]

2 Claims

1. An in vitro glycosylation method, comprising the steps of:

(i) transforming or transducing a host cell with a polynucleotide encoding a glycosyltransferase, or with a recombinant expression vector containing the polynucleotide;

(ii) culturing the host cell in a culture medium, thereby expressing the glycosyltransferase;

(iii) isolating and purifying the glycosyltransferase from the culture medium or the host cell; and

(iv) transferring a glycosyl group from a glycosyl donor to one of the following positions of a tetracyclic triterpenoid with the glycosyltransferase:

the first glycosyl group on position C20 and/or position C3 of the tetracyclic triterpenoid; and

thereby forming a glycosylated tetracyclic triterpenoid;

wherein the glycosyltransferase comprises SEQ ID NO:43 or a derivative polypeptide thereof; and wherein the derivative polypeptide, having glycosyltransferase activity, is formed by adding a tag sequence, a signal sequence or a secretion signal sequence to SEQ ID NO:43.