Industrial saponins

Lead Research Organisation: John Innes Centre
Department Name: Metabolic Biology

Abstract

The aim of this project is to bring saponin production into the domain of industrial biotechnology to make this diverse group of biosurfactants available for commercial exploitation at the required scale, structure specificity & cost, in the first instance for home & personal care (HPC) use. We have already demonstrated that saponins can synergise with conventional surfactants to reduce the total surfactant level required for cleaning performance in laundry detergents, and that such mixtures are particularly effective at low temperatures. Saponins are plant-derived triterpenoidal or steroidal surfactants. Saponins currently have limited commercial availability, being sourced from natural plant materials. This supply is deficient in scale, is of limited structure specificity and is too expensive for Unilever's intended use in laundry detergents. Unilever's concentrated detergents business alone would require 100 tonnes pa of saponin at 0.1% level and 20% product range penetration. Leading saponin supplier Desert King produces only 20 tonnes active saponin pa from natural extracts. Furthermore, we have demonstrated that certain saponin structures are significantly more effective than others. The current sources of saponins are natural extracts and comprise a heterogeneous mixture of saponin structures. Our IB innovation will enable considerable enrichment for the most effective saponin structures. Current commercial saponins are too expensive at $35/kg for crude extracts for use in anything other than premium products. We expect IB to be a more cost effective route to saponin supply.

Technical Summary

In a current project (TSB 131168), we have demonstrated production of tailored saponins in Nicotiana benthamiana and have expanded the synthetic biology tool kit genes and enzymes for tailored saponin bioengineering. However, this tobacco platform is not sufficiently scalable for commercial production. We need to transfer the saponin synthetic pathways into a microbial platform. We have selected yeast, Saccharomyces cerevisiae, as the most appropriate platform. Our aims are to push science and technology boundaries in multiple dimensions: (1) to transfer saponin metabolic pathways from model plant platform to yeast and engineer the pathway for commercially viable yield; although an academic proof of concept of simple monoglycosylated saponin production in yeast exists, further innovation is required to enable complex hyperglycosylated saponin production and to tackle yield & flux optimisation; (2) for downstream process development and scale up; (3) to further develop the triterpene toolbox of enzymes, genes and pathways for specific saponin structure enrichment & production; (4) to further understand the solution microstructure and surface adsorbed layer properties of mixtures of saponin & conventional surfactants, and (5) to continue to develop tangible benefits and structure-function relationships demonstrated in TSB 131168 for inclusion of saponins in model HPC formulations, extending the complexity of the models to include associated detergent technologies such as enzymes & dispersants. The above innovations will collectively enable a powerful business case for saponin commercialisation to be constructed.

Planned Impact

As described in proposal submitted to IUK.

Publications

10 25 50
 
Title Image of Nicotiana benthamiana 
Description Nicotiana benthamiana, a relative of the tobacco plant, is commonly used in plant research. Here, the leaf is used as a host organism to produce components of avenacin, a fluorescent antimicrobial produced by oat roots, which protects the roots from soil pathogens. A cluster of genes - the Sad genes - have been identified in oat as the instructions used to produce avenacin. The blue spots indicate the presence of parts of the avenacin compound introduced by inserting some of the Sad genes into the host plant. The other spots are controls to test the expression system. Image supplied by Aymeric Leveau, Osbourn laboratory. 
Type Of Art Image 
Year Produced 2015 
Impact Image placed on the Norwich Research Park Image Library, for free sharing and dissemination. 
URL http://images.norwichresearchpark.ac.uk/imagedetails.aspx?imgid=103
 
Title Winner of Norwich Research Park Image Library Competition: Dr Thomas Louveau 
Description More than 150 images of work were submitted by researchers, clinicians and scientists who work at the site, for the NRP image library competition. These were narrowed down to 12 finalists - which will form part of a calendar - from which an overall winner was selected. The winning image, entitled A New Planet, was taken by Thomas Louveau, a post-doctoral scientist in the Department of Metabolic Biology at the John Innes Centre. The image show a dried plant extract in a round flask taken from behind on a dark background. The extraction is part of the purification process of new-to-nature metabolites produced by synthetic biology approaches. The new metabolic pathway has been engineered in Nicotiana benthamiana, a relative of tobacco by scientists at the John Innes Centre on the Norwich Research Park who are studying natural products.. 
Type Of Art Image 
Year Produced 2015 
Impact A calendar was produced from the 12 final images, with "A New Planet" on the cover. The images are also displayed in the Centrum building on the Norwich Research Park. 
URL http://edp24.co.uk/news/tech/sheer_beauty_of_science_showcased_in_competition_at_norwich_research_pa...
 
Description Plants rich in saponins have been used for millennia as natural cleansing agents. The saponins are a structurally diverse group of glycosylated triterpenoids and sterols. In a previous TSB-funded project, we mapped structure-function relationships of triterpenoid saponins. Within this project we are continuing the development of bioengineered saponins for commercial use in the Home and Personal Care (HPC) industry. Specifically, we are: (1) Building a platform for saponin production which could be exploited commercially and demonstrate at pilot scale; (2) Further expanding the synthetic biology toolkit to increase the diversity of saponins that can be produced; (3) Further investigating the physical interactions of saponins with conventional surfactant systems to develop saponin formulations that can offer superiority over current HPC products; (4) Mitigating safety risks by understanding the structure-property rules which influence toxicological potential of some saponins.
Exploitation Route Saponins are soap-like substances or surfactants produced by certain plants. They have huge potential as a natural, biobased alternative to petrochemical surfactants for use in detergent products. However, extraction from plants is not economically feasible for use in detergents or sustainable on the basis of land use. This project sets out to establish a sustainable, commercially viable supply chain for the production of saponins in yeast. It tackles complex challenges of developing a yeast strain to produce the saponin at sufficient yield and how to recover the saponin from the fermentation medium. The project will also explore the physical properties of saponins alone and in mixtures with conventional surfactants, and will establish how best to formulate saponins into commercial detergent products for both economy and end results. Finally, the project will explore the safety of saponins using risk assessment methodology and in vitro methods. Promising findings generated within this project will be taken forward by our industrial partners for evaluation for potential commercialisation. A collaboration agreement with mutually beneficial terms and conditions is in place to enable this.
Sectors Chemicals,Energy,Environment,Manufacturing, including Industrial Biotechology

URL https://www.jic.ac.uk/directory/anne-osbourn/
 
Description Saponins are soap-like substances or surfactants produced by certain plants. They have huge potential as a natural, biobased alternative to petrochemical surfactants for use in detergent products. However, extraction from plants is not economically feasible for use in detergents or sustainable on the basis of land use. This project sets out to establish a sustainable, commercially viable supply chain for the production of saponins in yeast. It tackles complex challenges of developing a yeast strain to produce the saponin at sufficient yield and how to recover the saponin from the fermentation medium. The project will also explore the physical properties of saponins alone and in mixtures with conventional surfactants, and will establish how best to formulate saponins into commercial detergent products for both economy and end results. Finally, the project will explore the safety of saponins using risk assessment methodology and in vitro methods. Promising findings generated within this project will be taken forward by our industrial partners for evaluation for potential commercialisation. A collaboration agreement with mutually beneficial terms and conditions is in place to enable this.
First Year Of Impact 2015
Sector Agriculture, Food and Drink,Chemicals,Energy,Environment,Manufacturing, including Industrial Biotechology
Impact Types Societal,Economic

 
Description Built new image library for the Norwich Research Park 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact To help raise awareness of the high quality research carried out across the Norwich Research Park, we developed an open access image library to showcase images of NRP science accompanied by accessible legends suitable for a general audience. The image library has the potential to attract new traffic to the NRP institute websites and research group homepages and will allow groups such as the media, schools, the general public and other researchers to gain a glimpse of the great depth and diversity of research that is being carried out in Norwich. Although all the images are freely available to download and use, people are required to sign up to use the library and are asked to enter information at the time of download about their intended use of the image. This helps to capture who is using the library and where the images are being used. So far images have been downloaded 582 times and have been used in presentations, publications, on websites, for education and advertising. We ran an image competition to raise awareness across the NRP site and then assembled a judging panel to select the top 12 images which were used to create an NRP calendar for 2016. The calendar was sent to politicians, business leaders, industry and academics at institutions in the UK and internationally. The competition featured in the local paper, the Eastern Daily Press and an overall winner was picked and used to make a large canvas which is hanging in the new Centrum building. We have received many positive comments about the libraries ease of use for finding good quality, copyright free images to use in presentations from fellow researchers across the site. In October 2016 we put on a two-week, large scale image exhibition at the Forum in Norwich as part of the first Norwich Science Festival. The images attracted a lot of interest and several artists have been in touch with scientists whose research images were on display to set up new collaborations using science to inform artistic practice. We will post outcomes of these endeavours onto the image library website to widen our exposure to new audiences.
Year(s) Of Engagement Activity 2015,2016
URL http://images.norwichresearchpark.ac.uk/
 
Description Great British Bioscience Festival 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact We were successful in a winning a BBSRC science communication award to develop a new science stand called 'Nature's Factories' to run at local events and as part of the BBSRC's Great British Bioscience Festival showcasing the best of British bioscience in its 20th anniversary year.
Human evolution is tightly linked to our use of plants for food, building materials, fuel, medicines etc., and we continue to look for innovative ways to use natural resources to provide us with sustainable solutions that support our lifestyles. Scientific research is enabling us to discover and develop new plant products that improve our lifestyles by creating better medicines, healthier foods and greener technologies whilst also demonstrating the importance of protecting plant species diversity and ecosystems.
The Nature's Factories stand was designed to enable the public to find out how science is exploring and exploiting the valuable variety of chemicals made by plants as well as making natural remedies to take away and pick leaves from our fact-tree.

We took the stand to the Science in Norwich Day at the Forum in Norwich (1st June 2014), to the Cambridge Botanic Gardens (19th August 2014) and on the 12th November 2014 used the stand for a training workshop for EU scientists to learn how to create interactive displays for public events. On 14-16 November 2014, our exhibit headed to Museum Gardens, Bethnal Green, London to be showcased at BBSRC's Great British Bioscience Festival. The festival, delivered in partnership with London Science Festival was open to the general public and free to attend.
Since then the exhibit has featured at the Bury St Edmunds family science festival (21st March 2015), the Fascination of Plants day at the John Innes Centre (14th May 2015), the Festival of Plants at Cambridge Botanic Gardens (16th May 2015) and at the Youth Stem Awards (13th January 2016). We plan to use the exhibit throughout 2016.
Year(s) Of Engagement Activity 2014,2015,2016
URL http://www.bbsrc.ac.uk/engagement/exhibitions/gb-bioscience-festival/
 
Description High Value Chemicals from Plants- Harnessing the potential of synthetic biology for industrial biotechnology. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact High Value Chemicals from Plants- Harnessing the potential of synthetic biology for industrial biotechnology. 13-14 July 2015, Dunston Hall, Norwich. I delivered a 20min talk about the use of the HyperTrans system in goldengate vectors and its benefits for metabolic engineering.
Year(s) Of Engagement Activity 2015
 
Description NRP DTP Summer Conference 2015 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Professional Practitioners
Results and Impact NRP DTP Summer Conference 2015 held in the assembly house in Norwich. 15 min talk: Pathway engineering using the GoldenGate MoClo system.

Norwich Research Park Doctoral Training Partnerships
Year(s) Of Engagement Activity 2015
URL http://www.biodtp.norwichresearchpark.ac.uk/
 
Description Norwich Science Festival John Innes Centre Exhibit 22.10.16 
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 Norwich Science Festival.

The interactive 'Power of Plants' stand from the Latitude Festival made another appearance at the first Norwich Science Festival with Katrin Geisler coming over from Cambridge University to join JIC scientists Michael Stephenson, Hannah Griffiths, Colette Matthewman, Zhenhua Liu, Nadiatul Radzman, Dorota Jakubczyk, Don Nguyen, Miriam Walden, Jenni Rant and Roger Castells-Graells for two days as part of the Norwich Research Park's 'solving problems with science' weekend. A new addition to the stand was developed by Roger who introduced visitors to the structure of viruses through his challenge to build a giant virus particle. Plants are being used to produce virus particles for use as vaccines and so for visitors this deeper look at viruses in combination with a hands-on experience of mock infiltration of Nicotiana benthamiana plants with water enabled the scientists to explain the techniques involved and exactly what is being made by plants in this system. People were very interested in the variety of science on display and asked lots of questions. It was good to showcase our research at the first Norwich Science Festival and to show people locally how far reaching the work taking place in essentially a rural county can be in terms of impact to societies across the globe.
Year(s) Of Engagement Activity 2016
URL http://norwichsciencefestival.co.uk/
 
Description Science in Norwich day 
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 Helped Jenni Rant (SAW) to hold a stand describing the scientific research ongoing at JIC, at the Science in Norwich Day at the Forum on the 1st of June 2014 .
Year(s) Of Engagement Activity 2014
URL http://soin.org.uk/2014/04/science-in-norwich-day-2014/
 
Description Science, Art and Writing (SAW) project on plant natural products, Suffolk 
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 A Science, Art and Writing (SAW) project on plant natural products was run for 90 students at St Benedicts Secondary School, Suffolk, UK on 3 July 2015. The students collected plant samples from the school grounds and used them in chromatography experiments to see how many different types of pigments they could extract.
Year(s) Of Engagement Activity 2015
 
Description Submitted scientific images to the Norwich Research Park image library 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact The NRP image library consists of images arising from research in life and environmental sciences on the Norwich Research Park . The aim of NRP research is to deliver solutions to the global challenges of healthy ageing, food and energy security, sustainability and environmental change and this is reflected in the images in the library.
Year(s) Of Engagement Activity 2016
URL http://images.norwichresearchpark.ac.uk/searchresults.aspx?query=leveau