Engineering Quillaja saponin biosynthesis pathways for bio-production of QS-21
Lead Research Organisation:
John Innes Centre
Department Name: Metabolic Biology
Abstract
Plants are a rich source of drugs and other high-value compounds. Morphine was the first plant drug to be purified and sold commercially and salicylic acid the first to be chemically synthesized. However, despite these and other successes, the vast majority of plant metabolic diversity has remained untapped due to the problems of accessing source species, purifying compounds and the challenges of chemical synthesis. We have established transient plant expression technology for the rapid reconstruction of plant metabolic pathways and demonstrated that this system can be used to access gram-scale quantities of products. This technology is readily scalable and has been used for commercial production of vaccines. The engineering and production of the highly potent Quillaja adjuvant QS-21 using the Hypertrans system will provide a flagship proof of concept to demonstrate the power of this platform for accessing high-value products from plants and pave the way for a new commercial venture.
Organisations
People |
ORCID iD |
| Anne Osbourn (Principal Investigator) |
Publications
Lee N
(2020)
The Global Garden project: Imagining plant science
in PLANTS, PEOPLE, PLANET
Louveau T
(2019)
The Sweet Side of Plant-Specialized Metabolism.
in Cold Spring Harbor perspectives in biology
Orme A
(2019)
A noncanonical vacuolar sugar transferase required for biosynthesis of antimicrobial defense compounds in oat.
in Proceedings of the National Academy of Sciences of the United States of America
Polturak G
(2022)
New and emerging concepts in the evolution and function of plant biosynthetic gene clusters
in Current Opinion in Green and Sustainable Chemistry
Reed J
(2018)
Engineering terpenoid production through transient expression in Nicotiana benthamiana.
in Plant cell reports
Stephenson M
(2020)
Making drugs out of sunlight and 'thin air': an emerging synergy of synthetic biology and natural product chemistry
in The Biochemist
Stephenson M
(2020)
Comprehensive Natural Products III
| Description | Advice for Government Report on Synthetic Cell. |
| Geographic Reach | National |
| Policy Influence Type | Contribution to new or Improved professional practice |
| Description | Attended Roundtable meeting to discuss how Government might further help in the UK's engineering biology sector. |
| Geographic Reach | National |
| Policy Influence Type | Contribution to a national consultation/review |
| Description | BBSRC Global Carbon for Manufacturing Working Group |
| Geographic Reach | Multiple continents/international |
| Policy Influence Type | Participation in a guidance/advisory committee |
| Description | Visit by Minister of State in the Department for Science, Innovation and Technology |
| Geographic Reach | National |
| Policy Influence Type | Participation in a guidance/advisory committee |
| Description | Engineering saponin biosynthesis pathways for bio-production of novel vaccine adjuants |
| Amount | £646,458 (GBP) |
| Funding ID | BB/W017857/1 |
| Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
| Sector | Public |
| Country | United Kingdom |
| Start | 05/2022 |
| End | 04/2024 |
| Title | Transient plant expression technology for triterpene production at preparative scale |
| Description | We have develop a method for transient heterologous expression of biosynthetic enzymes in N. benthamiana for production of high-value triterpenes. Agro-infiltration is an efficient and simple means of achieving transient expression in N. benthamiana. The process involves infiltration of plant leaves with a suspension of Agrobacterium tumefaciens carrying the expression construct(s) of interest. Co-infiltration of an additional A. tumefaciens strain carrying an expression construct encoding an enzyme that boosts precursor supply significantly increases yields. After a period of five days, the infiltrated leaf material can be harvested and processed to extract and isolate the resulting triterpene product(s). This is a process that is linearly and reliably scalable, simply by increasing the number of plants used in the experiment. We have developed a protocol for rapid preparative-scale production of triterpenes utilizing this plant-based platform. The protocol utilizes an easily replicable vacuum infiltration apparatus, which allows the simultaneous infiltration of up to four plants, enabling batch-wise infiltration of hundreds of plants in a short period of time. |
| Type Of Material | Technology assay or reagent |
| Year Produced | 2017 |
| Provided To Others? | Yes |
| Impact | Generation of gram-scale quantities of 98% pure triterpenes and demonstration that we can rapidly carry out combinatorial expression of enzymes from our triterpene toolkit to generate known and new-to-nature compounds. This had attracted considerable interest from industry and led to four new projects directly funded by different companies in the pharma, ag, food and drink and home and personal care sectors. |
| URL | https://www.jove.com/video/58169/transient-expression-nicotiana-benthamiana-leaves-for-triterpene |
| Description | Activity stand at mixed arts festival |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Public/other audiences |
| Results and Impact | We put together an activity for the science marquee at the Latitude Festival, working with colleagues from the University of East Anglia, Food and Farming Discovery Trust, CEFAS, Precious Plastics East and some local artists with the overall topic of microplastics in the environment. Some of the activity was focused on the problems microplastics cause as pollutants in the ocean and in food chains but we also had a section on plastics in our daily lives that enabled exploration of alternative plant-based materials and the potential for greener alternatives from both biological engineering for new materials and better recycling and remaking processes. We feel that by looking at a problem (microplastics in our environment) but then exploring better alternatives we conveyed a sense of agency to the families to seek out and support alternatives in their daily lives. We were particularly pleased to hear many people pledge to give up using clingfilm in favour of beeswax and paper alternatives to wrap their lunch! |
| Year(s) Of Engagement Activity | 2022 |
| Description | Biodiversity Workshop |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | New SAW workshop delivered at Bignold Primary School on World Biodiversity Day to introduce year 4 children to the importance of biodiversity for people and the planet and to some of the factors that are causing species decline across the globe. The children were amazed by the wealth of species, their importance in the ecosystem and how humans utilise natural products and shocked by the driving forces to extinction, a term they believed to only apply in the past, linked to dinosaurs, and not something that was ongoing. |
| Year(s) Of Engagement Activity | 2018 |
| URL | https://www.sawtrust.org/in-schools/world-biodiversity-day-at-bignold-primary-school/ |
| Description | Global garden |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Public/other audiences |
| Results and Impact | Following on from the pilot of the Global Garden workshop in 2017 we ran a public workshop as part of the Norwich Science Festival that invited anyone to explore plant natural products and the regulations and ethics around access and benefit sharing. We then ran another session for a group of scientists which provided an interesting contrast of opinions around ethics compared to those shared by members of the public. This is an ongoing project that seeks to raise awareness of these issues as well as to get people thinking and talking about the importance of plants. In 2019 we took the Global Garden workshop to Cambridge as part of the Festival of Ideas and ran the workshop at the Cambridge Botanic Gardens. We also had garden curator Dr Sam Brockington attend and give a presentation on the importance of plant collections for research as well as for enjoyment. |
| Year(s) Of Engagement Activity | 2017,2018,2019 |
| Description | Royal Norfolk Show plant science stand |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Schools |
| Results and Impact | We designed and delivered science activities in the Discovery Zone at the Royal Norfolk Show for visiting children to learn about the properties of gluten in wheat and the use of insect predators as biocontrol for crop protection. Hundreds of children participated in our activities and most had heard of gluten as it is commonly mentioned now in terms of food allergies so we were able to give them some context around what gluten actually is, where its found and the properties it has. Many children were surprised by this! |
| Year(s) Of Engagement Activity | 2019 |
| Description | SAW Seminar - Sowing the seeds for science outreach |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Professional Practitioners |
| Results and Impact | Presented creative outreach projects and ways to capture impact at the department seminar to encourage scientists to be bold with their outreach plans and to demonstrate the value of documenting the process and the outcomes for reporting, reflection and to improve the method. |
| Year(s) Of Engagement Activity | 2020 |
| Description | Science Art and Writing (SAW) Project in school |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | Delivered a SAW project on the theme of plants at Tacolneston Primary, a very rural school in Norfolk. The school were studying Arctic and Antarctic biomes and so we worked with them to add some plant stories to the very zoocentric approach schools usually take when exploring these regions as habitats. This enabled us to explain adaptations to environments which are particularly fascinating in plants and we touched on some local work by colleagues in Cambridge who discovered alpine plants making a fine wool out of flavonoids to cover its leaves. We explored chemical diversity and its many uses in plants and then looked at chemicals in plants that are useful to humans. The children were surprised to see this dynamic side of plants. We then focused on colours, flavours and fragrances and made extractions and then altered our extracts with acids and bases to make many new versions. The children enjoyed this plant-based chemistry session and then took their new facts, vocabulary and concepts forward to write poetry and create art to explore the topic further. |
| Year(s) Of Engagement Activity | 2023 |
| Description | Talking Plants Conference |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Professional Practitioners |
| Results and Impact | Delivered a workshop at the Talking Plants Conference at Cambridge Botanic Garden. This conference takes place at a different botanic garden nationally each year and brings together garden staff and other organisations that work with plants and the public. Our workshop showcased some of the best plant-themed outreach projects we have delivered to share ideas and activities that provide a platform to engage people with plants. |
| Year(s) Of Engagement Activity | 2018 |
| URL | https://bgen.org.uk/conferences/talking-plants-2018-conference-proceedings/ |
| Description | aBIOTECH Virtual Seminar by Professor Anne Osbourn. Title of talk: 'Harnessing plant metabolic diversity'. Zoom meeting. |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | "aBIOTECH Virtual Seminar": Zoom Presentation: Presenter: Professor Anne Osbourn (John Innes Centre) at the invitation of Professor Xiaoquan Qi (CAS Institute of Botany, Beijing, China). Title of talk: "Harnessing plant metabolic diversity" JIC organiser: Dr Jie Li. Date and time: 10:00-11:30 (UK time), 24 February 2023. Audience of approximately 1000 people. Hosted by aBIOTECH editor in chief Professor Sanwen Huang. Keywords Notes |
| Year(s) Of Engagement Activity | 2023 |