Title:
GENETICALLY ENGINEERED BACTERIA AND METHODS FOR PREPARING A FUCOSYLATED OLIGOSACCHARIDE USING THE SAME
Document Type and Number:
WIPO Patent Application WO/2023/098299
Kind Code:
A1
Abstract:
The invention discloses a genetically engineered bacterium and a method for preparing a fucosylated oligosaccharide using the same. The method comprises: transferring a fucosyl group of a donor to an oligosaccharide receptor by a fucosyltransferase heterologously expressed in a genetically engineered bacterium; wherein the donor is a nucleotide-activated donor, the fucosyltransferase has α-1, 2-fucosyltransferase activity; wherein, the fucosyltransferase is selected from one or more of the enzymes corresponding to NCBI Accession Numbers WP_109047124.1, RTL12957.1, MBP7103497.1, WP_120175093.1, RYE22506.1, WP_140393075.1 and HJB91111.1. The preparation method of the invention has high yield, greatly improved substrate conversion rate and product conversion rate, and has the potential to be applied to industrial production.
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Inventors:
CHENG ZHANBING (CN)
LIU YUROU (CN)
JIAO QI (CN)
XU HONG (CN)
TIAN ZHENHUA (CN)
LIU YUROU (CN)
JIAO QI (CN)
XU HONG (CN)
TIAN ZHENHUA (CN)
Application Number:
PCT/CN2022/124634
Publication Date:
June 08, 2023
Filing Date:
October 11, 2022
Export Citation:
Assignee:
SYNAURA BIOTECHNOLOGY SHANGHAI CO LTD (CN)
International Classes:
C12P19/00; C12N9/10; C12P19/18; C12P19/26
Domestic Patent References:
| WO2015175801A1 | 2015-11-19 | |||
| WO2014018596A2 | 2014-01-30 |
Foreign References:
| EP3425052A1 | 2019-01-09 |
Other References:
DATABASE NCBI [online] NIH (USA); 22 May 2018 (2018-05-22), ANONYMOUS: "alpha-1,2-fucosyltransferase [Azospirillum sp. TSA6c]", XP093017314, retrieved from https://www.ncbi.nlm.nih.gov/protein/WP_109047124 Database accession no. WP_109047124
DATABASE GenBank [online] NIH; 22 December 2018 (2018-12-22), KANTOR R S ET AL: "MAG: alpha-1,2-fucosyltransferase [Neisseriaceae bacterium]", XP093017320, Database accession no. RTL12957.1
DATABASE GenBank [online] 15 April 2021 (2021-04-15), SCHNEIDER D: "MAG: alpha-1,2-fucosyltransferase [Bacteroidales bacterium]", XP093017326, retrieved from NIH Database accession no. MBP7103497.1
DATABASE GenBank [online] NIH; 6 February 2019 (2019-02-06), CROMBIE ANDREW T.: "alpha-1,2-fucosyltransferase [Sphingobacteriaceae bacterium]", XP093017328, retrieved from https://www.ncbi.nlm.nih.gov/protein/RYE22506 Database accession no. RYE22506.1
DATABASE NCBI [online] NIH; 4 October 2018 (2018-10-04), ANONYMOUS: "alpha-1,2-fucosyltransferase [Prevotella melaninogenica]", XP093017335, retrieved from https://www.ncbi.nlm.nih.gov/protein/WP_120175093 Database accession no. WP_120175093.1
DATABASE NCBI [online] NIH; 6 October 2020 (2020-10-06), ANONYMOUS: "alpha-1,2-fucosyltransferase [Lachnoclostridium sp. An138]", XP093017337, retrieved from https://www.ncbi.nlm.nih.gov/protein/WP_140393075 Database accession no. WP_140393075.1
DATABASE Genbank [online] NIH; 5 July 2021 (2021-07-05), GILROY R: "MAG TPA: alpha-1,2-fucosyltransferase [Candidatus Eisenbergiella merdigallinarum]", XP093017342, retrieved from https://www.ncbi.nlm.nih.gov/protein/HJB91111 Database accession no. HJB91111.1
"NCBI", Database accession no. WP_ 109047124.1
QI LIBINGBING SUNJUN CHENYIWEN ZHANGYU JIANGSHENG YANG: "A modified pCas/pTargetF system for CRISPR-Cas9-assisted genome editing in Escherichia coli", ACTA BIOCHIMICA ET BIOPHYSICA SINICA, vol. 53, May 2021 (2021-05-01), pages 620 - 627
DUMON, C.PRIEM, B.MARTIN, S.L. ET AL.: "In vivo fucosylation of lacto-N-neotetraose and lacto-N-neohexaose by heterologous expression of Helicobacter pylori a-1,3 fucosyltransferase in engineered Escherichia coli", GLYCOCONJ J, vol. 18, 2001, pages 465 - 474, XP002380631, DOI: 10.1023/A:1016086118274
DATABASE GenBank [online] NIH; 22 December 2018 (2018-12-22), KANTOR R S ET AL: "MAG: alpha-1,2-fucosyltransferase [Neisseriaceae bacterium]", XP093017320, Database accession no. RTL12957.1
DATABASE GenBank [online] 15 April 2021 (2021-04-15), SCHNEIDER D: "MAG: alpha-1,2-fucosyltransferase [Bacteroidales bacterium]", XP093017326, retrieved from NIH Database accession no. MBP7103497.1
DATABASE GenBank [online] NIH; 6 February 2019 (2019-02-06), CROMBIE ANDREW T.: "alpha-1,2-fucosyltransferase [Sphingobacteriaceae bacterium]", XP093017328, retrieved from https://www.ncbi.nlm.nih.gov/protein/RYE22506 Database accession no. RYE22506.1
DATABASE NCBI [online] NIH; 4 October 2018 (2018-10-04), ANONYMOUS: "alpha-1,2-fucosyltransferase [Prevotella melaninogenica]", XP093017335, retrieved from https://www.ncbi.nlm.nih.gov/protein/WP_120175093 Database accession no. WP_120175093.1
DATABASE NCBI [online] NIH; 6 October 2020 (2020-10-06), ANONYMOUS: "alpha-1,2-fucosyltransferase [Lachnoclostridium sp. An138]", XP093017337, retrieved from https://www.ncbi.nlm.nih.gov/protein/WP_140393075 Database accession no. WP_140393075.1
DATABASE Genbank [online] NIH; 5 July 2021 (2021-07-05), GILROY R: "MAG TPA: alpha-1,2-fucosyltransferase [Candidatus Eisenbergiella merdigallinarum]", XP093017342, retrieved from https://www.ncbi.nlm.nih.gov/protein/HJB91111 Database accession no. HJB91111.1
"NCBI", Database accession no. WP_ 109047124.1
QI LIBINGBING SUNJUN CHENYIWEN ZHANGYU JIANGSHENG YANG: "A modified pCas/pTargetF system for CRISPR-Cas9-assisted genome editing in Escherichia coli", ACTA BIOCHIMICA ET BIOPHYSICA SINICA, vol. 53, May 2021 (2021-05-01), pages 620 - 627
DUMON, C.PRIEM, B.MARTIN, S.L. ET AL.: "In vivo fucosylation of lacto-N-neotetraose and lacto-N-neohexaose by heterologous expression of Helicobacter pylori a-1,3 fucosyltransferase in engineered Escherichia coli", GLYCOCONJ J, vol. 18, 2001, pages 465 - 474, XP002380631, DOI: 10.1023/A:1016086118274
Attorney, Agent or Firm:
LIU, SHEN & ASSOCIATES (CN)
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