PMID- 35655274
OWN - NLM
STAT- MEDLINE
DCOM- 20220606
LR  - 20220716
IS  - 1475-2859 (Electronic)
IS  - 1475-2859 (Linking)
VI  - 21
IP  - 1
DP  - 2022 Jun 2
TI  - Engineered Bacillus subtilis for the de novo production of 2'-fucosyllactose.
PG  - 110
LID - 10.1186/s12934-022-01838-w [doi]
LID - 110
AB  - BACKGROUND: The most abundant human milk oligosaccharide in breast milk, 
      2'-fucosyllactose (2'-FL), has been approved as an additive to infant formula due 
      to its multifarious nutraceutical and pharmaceutical functions in promoting 
      neonate health. However, the low efficiency of de novo synthesis limits the 
      cost-efficient bioproduction of 2'-FL. RESULTS: This study achieved 2'-FL de novo 
      synthesis in a generally recognized as safe (GRAS) strain Bacillus subtilis. 
      First, a de novo biosynthetic pathway for 2'-FL was introduced by expressing the 
      manB, manC, gmd, wcaG, and futC genes from Escherichia coli and Helicobacter 
      pylori in B. subtilis, resulting in 2'-FL production of 1.12 g/L. Subsequently, a 
      2'-FL titer of 2.57 g/L was obtained by reducing the competitive lactose 
      consumption, increasing the regeneration of the cofactor 
      guanosine-5'-triphosphate (GTP), and enhancing the supply of the precursor 
      mannose-6-phosphate (M6P). By replacing the native promoter of endogenous manA 
      gene (encoding M6P isomerase) with a constitutive promoter P7, the 2'-FL titer in 
      shake flask reached 18.27 g/L. The finally engineered strain BS21 could produce 
      88.3 g/L 2'-FL with a yield of 0.61 g/g lactose in a 3-L bioreactor, without the 
      addition of antibiotics and chemical inducers. CONCLUSIONS: The efficient de novo 
      synthesis of 2'-FL can be achieved by the engineered B. subtilis, paving the way 
      for the large-scale bioproduction of 2'-FL titer in the future.
CI  - (c) 2022. The Author(s).
FAU - Zhang, Quanwei
AU  - Zhang Q
AD  - Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of 
      Education, Jiangnan University, Wuxi, 214122, China.
AD  - Science Center for Future Foods, Jiangnan University, Wuxi, 214122, China.
FAU - Liu, Zhenmin
AU  - Liu Z
AD  - State Key Laboratory of Dairy Biotechnology, Shanghai Engineering Research Center 
      of Dairy Biotechnology, Dairy Research Institute, Bright Dairy & Food Co., Ltd, 
      Shanghai, 200436, China.
FAU - Xia, Hongzhi
AU  - Xia H
AD  - Nantong Licheng Biological Engineering Co., Ltd, Shanghai, 200000, China.
FAU - Huang, Ziyang
AU  - Huang Z
AD  - Yixing Institute of Food Biotechnology Co., Ltd, Yixing, 214200, China.
FAU - Zhu, Yonglian
AU  - Zhu Y
AD  - Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of 
      Education, Jiangnan University, Wuxi, 214122, China.
AD  - Science Center for Future Foods, Jiangnan University, Wuxi, 214122, China.
FAU - Xu, Linfeng
AU  - Xu L
AD  - Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of 
      Education, Jiangnan University, Wuxi, 214122, China.
AD  - Science Center for Future Foods, Jiangnan University, Wuxi, 214122, China.
FAU - Liu, Yanfeng
AU  - Liu Y
AD  - Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of 
      Education, Jiangnan University, Wuxi, 214122, China.
AD  - Science Center for Future Foods, Jiangnan University, Wuxi, 214122, China.
FAU - Li, Jianghua
AU  - Li J
AD  - Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of 
      Education, Jiangnan University, Wuxi, 214122, China.
AD  - Science Center for Future Foods, Jiangnan University, Wuxi, 214122, China.
FAU - Du, Guocheng
AU  - Du G
AD  - Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of 
      Education, Jiangnan University, Wuxi, 214122, China.
AD  - Science Center for Future Foods, Jiangnan University, Wuxi, 214122, China.
FAU - Lv, Xueqin
AU  - Lv X
AD  - Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of 
      Education, Jiangnan University, Wuxi, 214122, China.
AD  - Science Center for Future Foods, Jiangnan University, Wuxi, 214122, China.
FAU - Liu, Long
AU  - Liu L
AD  - Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of 
      Education, Jiangnan University, Wuxi, 214122, China. longliu@jiangnan.edu.cn.
AD  - Science Center for Future Foods, Jiangnan University, Wuxi, 214122, China. 
      longliu@jiangnan.edu.cn.
LA  - eng
GR  - 31930085, 31870069/National Natural Science Foundation of China/
GR  - SKLF-ZZB-202106, JUSRP121010/Fundamental Research Funds for the Central 
      Universities/
GR  - 2018YFA0900504, 2018YFA0900300/National Key Research and Development Program of 
      China/
PT  - Journal Article
DEP - 20220602
PL  - England
TA  - Microb Cell Fact
JT  - Microbial cell factories
JID - 101139812
RN  - 0 (Trisaccharides)
RN  - EC 2.4.1.- (Fucosyltransferases)
RN  - J2B2A4N98G (Lactose)
RN  - XO2533XO8R (2'-fucosyllactose)
SB  - IM
MH  - *Bacillus subtilis/genetics/metabolism
MH  - Escherichia coli/metabolism
MH  - Fucosyltransferases/metabolism
MH  - Humans
MH  - Infant, Newborn
MH  - *Lactose/metabolism
MH  - Trisaccharides
PMC - PMC9164505
OTO - NOTNLM
OT  - 2'-fucosyllactose
OT  - Bacillus subtilis
OT  - De novo pathway
OT  - GDP-L-fucose
COIS- The authors declare no competing financial interest.
EDAT- 2022/06/03 06:00
MHDA- 2022/06/07 06:00
PMCR- 2022/06/02
CRDT- 2022/06/02 23:46
PHST- 2022/05/04 00:00 [received]
PHST- 2022/05/26 00:00 [accepted]
PHST- 2022/06/02 23:46 [entrez]
PHST- 2022/06/03 06:00 [pubmed]
PHST- 2022/06/07 06:00 [medline]
PHST- 2022/06/02 00:00 [pmc-release]
AID - 10.1186/s12934-022-01838-w [pii]
AID - 1838 [pii]
AID - 10.1186/s12934-022-01838-w [doi]
PST - epublish
SO  - Microb Cell Fact. 2022 Jun 2;21(1):110. doi: 10.1186/s12934-022-01838-w.