Metabolic engineering of Priestia megaterium for 2’-fucosyllactose production
Abstract Background 2′-Fucosyllactose (2′-FL) is a predominant human milk oligosaccharide that significantly enhances infant nutrition and immune health. This study addresses the need for a safe and economical production of 2’-FL by employing Generally Recognized As Safe (GRAS) microbial strain, Pri...
Saved in:
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2025-01-01
|
| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12934-024-02620-w |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1841559018087120896 |
|---|---|
| author | Bu-Soo Park Jihee Yoon Jun-Min Lee Sang-Hyeok Cho Yoojeong Choi Byung-Kwan Cho Min-Kyu Oh |
| author_facet | Bu-Soo Park Jihee Yoon Jun-Min Lee Sang-Hyeok Cho Yoojeong Choi Byung-Kwan Cho Min-Kyu Oh |
| author_sort | Bu-Soo Park |
| collection | DOAJ |
| description | Abstract Background 2′-Fucosyllactose (2′-FL) is a predominant human milk oligosaccharide that significantly enhances infant nutrition and immune health. This study addresses the need for a safe and economical production of 2’-FL by employing Generally Recognized As Safe (GRAS) microbial strain, Priestia megaterium ATCC 14581. This strain was chosen for its robust growth and established safety profile and attributing suitable for industrial-scale production. Results The engineering targets included the deletion of the lacZ gene to prevent lactose metabolism interference, introduction of α-1,2-fucosyltransferase derived from the non-pathogenic strain, and optimization of the GDP-L-fucose biosynthesis pathway through the overexpression of manA and manC. These changes, coupled with improvements in lactose uptake and utilization through random mutagenesis, led to a high 2’-FL yield of 28.6 g/L in fed-batch fermentation, highlighting the potential of our metabolic engineering strategies on P. megaterium. Conclusions The GRAS strain P. megaterium ATCC 14581 was successfully engineered to overproduce 2’-FL, a valuable human milk oligosaccharide, through a series of genetic modifications and metabolic pathway optimizations. This work underscores the feasibility of using GRAS strains for the production of oligosaccharides, paving the way for safer and more efficient methods in biotechnological applications. Future studies could explore additional genetic modifications and optimization of fermentation conditions of the strain to further enhance 2’-FL yield and scalability. |
| format | Article |
| id | doaj-art-92f25fddad7c41599a90f28b5d5e08ba |
| institution | Kabale University |
| issn | 1475-2859 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj-art-92f25fddad7c41599a90f28b5d5e08ba2025-01-05T12:50:46ZengBMCMicrobial Cell Factories1475-28592025-01-0124111010.1186/s12934-024-02620-wMetabolic engineering of Priestia megaterium for 2’-fucosyllactose productionBu-Soo Park0Jihee Yoon1Jun-Min Lee2Sang-Hyeok Cho3Yoojeong Choi4Byung-Kwan Cho5Min-Kyu Oh6Department of Chemical & Biological Engineering, Korea UniversitySamyang Corp.Department of Chemical & Biological Engineering, Korea UniversityDepartment of Biological Sciences, Korea Advanced Institute of Science and TechnologyDepartment of Biological Sciences, Korea Advanced Institute of Science and TechnologyDepartment of Biological Sciences, Korea Advanced Institute of Science and TechnologyDepartment of Chemical & Biological Engineering, Korea UniversityAbstract Background 2′-Fucosyllactose (2′-FL) is a predominant human milk oligosaccharide that significantly enhances infant nutrition and immune health. This study addresses the need for a safe and economical production of 2’-FL by employing Generally Recognized As Safe (GRAS) microbial strain, Priestia megaterium ATCC 14581. This strain was chosen for its robust growth and established safety profile and attributing suitable for industrial-scale production. Results The engineering targets included the deletion of the lacZ gene to prevent lactose metabolism interference, introduction of α-1,2-fucosyltransferase derived from the non-pathogenic strain, and optimization of the GDP-L-fucose biosynthesis pathway through the overexpression of manA and manC. These changes, coupled with improvements in lactose uptake and utilization through random mutagenesis, led to a high 2’-FL yield of 28.6 g/L in fed-batch fermentation, highlighting the potential of our metabolic engineering strategies on P. megaterium. Conclusions The GRAS strain P. megaterium ATCC 14581 was successfully engineered to overproduce 2’-FL, a valuable human milk oligosaccharide, through a series of genetic modifications and metabolic pathway optimizations. This work underscores the feasibility of using GRAS strains for the production of oligosaccharides, paving the way for safer and more efficient methods in biotechnological applications. Future studies could explore additional genetic modifications and optimization of fermentation conditions of the strain to further enhance 2’-FL yield and scalability.https://doi.org/10.1186/s12934-024-02620-wMetabolic engineering2’-fucosyllactoseα-1,2-fucosyltransferaseHMOGRASPriestia megaterium ATCC 14581 |
| spellingShingle | Bu-Soo Park Jihee Yoon Jun-Min Lee Sang-Hyeok Cho Yoojeong Choi Byung-Kwan Cho Min-Kyu Oh Metabolic engineering of Priestia megaterium for 2’-fucosyllactose production Microbial Cell Factories Metabolic engineering 2’-fucosyllactose α-1,2-fucosyltransferase HMO GRAS Priestia megaterium ATCC 14581 |
| title | Metabolic engineering of Priestia megaterium for 2’-fucosyllactose production |
| title_full | Metabolic engineering of Priestia megaterium for 2’-fucosyllactose production |
| title_fullStr | Metabolic engineering of Priestia megaterium for 2’-fucosyllactose production |
| title_full_unstemmed | Metabolic engineering of Priestia megaterium for 2’-fucosyllactose production |
| title_short | Metabolic engineering of Priestia megaterium for 2’-fucosyllactose production |
| title_sort | metabolic engineering of priestia megaterium for 2 fucosyllactose production |
| topic | Metabolic engineering 2’-fucosyllactose α-1,2-fucosyltransferase HMO GRAS Priestia megaterium ATCC 14581 |
| url | https://doi.org/10.1186/s12934-024-02620-w |
| work_keys_str_mv | AT busoopark metabolicengineeringofpriestiamegateriumfor2fucosyllactoseproduction AT jiheeyoon metabolicengineeringofpriestiamegateriumfor2fucosyllactoseproduction AT junminlee metabolicengineeringofpriestiamegateriumfor2fucosyllactoseproduction AT sanghyeokcho metabolicengineeringofpriestiamegateriumfor2fucosyllactoseproduction AT yoojeongchoi metabolicengineeringofpriestiamegateriumfor2fucosyllactoseproduction AT byungkwancho metabolicengineeringofpriestiamegateriumfor2fucosyllactoseproduction AT minkyuoh metabolicengineeringofpriestiamegateriumfor2fucosyllactoseproduction |