Enhancing cobalamin (vitamin B12) bioavailability in culturally appropriate foods in India

Lead Research Organisation: University of Kent


B12 deficiency and insufficiency is thought to represent a hidden worldwide epidemic of nutrient malnutrition in both developed and developing countries. However, there is no doubt that in India the problem is on a vast scale, with estimates of around 1 billion people having low levels of the nutrient. This is a consequence not only of general malnutrition but also due to cultural practices associated with vegetarianism, since crops have very low levels of B12. In this application we aim to initiate a programme of work to address this nutrient-deficiency challenge, by combining expertise in the UK with that found in India. Specifically, we want to investigate if a newly constructed recombinant strain of E. coli can be used to provide large quantities of purified B12. To achieve this we will combine the metabolic engineering work at the University of Kent with the process engineering capability found at the ICT in Mumbai. The development of a suitable extraction procedure will allow us to interact with chemical companies such as Cipla, who have expressed an interest in taking this forward if a suitable high-yielding method is developed. This holds the potential for India to be able to produce all its own B12 rather than having to rely on expensive imports. In so doing, this will also address serious concerns about global nutrient supply, since currently around 90% of B12 is produced in China. We also plan to explore the use of alternative culturally acceptable enriched sources of B12 in the form of algae and higher plants. Many algae are known to be able to absorb the nutrient but only limited studies have been undertaken in order to assess whether the B12 found in algae is bioavailable to humans. Similarly, we have recently found that some higher plants, such as spring cress, are also able to absorb B12 when the plants are grown with the nutrient. However, as with algae, it is not known whether the nutrient is bioavailable. Therefore, apart from investigating ways to enhance B12 uptake into algae and higher plants, we will undertake volunteer trials to determine whether food prepared from these sources is able to alleviate B12 deficiency in the volunteers. This work will combine expertise in the UK in providing the growth conditions, the food technologists in Mumbai and the clinical expertise in Pune to address this problem. This represents an exciting series of experiments that holds the potential to provide transformative solutions to B12 deficiency in both developing developed countries.

Technical Summary

Vitamin B12 is a key nutrient that, like folic acid, plays a vital role in metabolism that affects cell growth and division and whose deficiency is manifested in forms of anaemia and cognitive impairment. As with folate it is also an independent risk factor in neural tube development. There are thus strong arguments to ensure that B12 levels are properly maintained through a sufficient diet, but as the nutrient is comparatively expensive it has not been considered for supplementation programmes. Moreover, in many developing countries, as a consequence of either cultural practices or because meat is expensive, large swathes of the population survive on vegetarian-rich diets, which are naturally low in vitamin B12. For these reasons developing countries have low levels of B12 with associated consequences for the health economy. In this project we aim to address the problem of B12 deficiency and insufficiency in India, a country that has the highest level of B12 deficiency in the world. Here, we plan to develop methods to enhance the extraction of B12 from a recombinant strain of E. coli in order that large-scale fermentative production of the nutrient can be considered in India. We also to plan to investigate a range of culturally-suitable food sources for B12 enrichment. Specifically, we plan to investigate the use of micro- and macro-algae for the uptake of B12 and to use these as fortified nutrient supplements in human trials. Similarly, we have recently shown that some higher plants such as garden cress can also accumulate B12 when grown in the presence of the nutrient. We therefore plan to investigate whether other plants can also accumulate B12 and again determine whether the B12 in these plants is bioavailable to humans. By combining the expertise in India at the ICT in Mumbai and the clinicians at the KEM hospital in Pune we will initiate trials with suitable volunteers from B12-deficient areas in order to study the bioavailability of B12 from such sources.

Planned Impact

The research described in this application will have a major impact on several areas of science, including nutrition, synthetic ecology, and metabolic engineering. The research relates to how engineered cells can be used to help in the overproduction of a complex small molecule, vitamin B12 and how this nutrient can be efficiently extracted from cultures. Specifically, this research holds the potential to generate a safe, reliable and affordable source of the nutrient for both industry and humanitarian causes. A cheap source of B12 could potentially improve the lives of tens of millions of people worldwide if it is employed in a supplementation programme.

The research falls well within the remit of nutrition and health and is therefore addressing a key priority area. Apart from using engineered systems for B12 production, the application also aims to study the use of alternative culturally-acceptable vegetarian sources of the nutrient. Specifically, the applications aims to explore the use of algae and higher plants as enriched food sources for the nutrient after growth in the presence of B12. A key outcome of this study will be the determination as to whether the B12 absorbed in these systems in bioavailable to humans.

The beneficiaries of this research will be researchers in academia and industry who are interested in health and nutrition and the global supply of nutrients. There is a current strong interest in this area and science needs to put forward a strong representation in terms of the positive contributions that it can make to ensure that supplies are not dominated by one single country. The research will not only provide essential information about uptake of nutrients can be enhanced, but it will also provide greater insight into the provision and procurement in complex communities. It will demonstrate how cells can be optimised to resource their nutrient components. We will ensure that our findings are widely disseminated through oral communication, research publications and reviews, and press releases. Specifically, our research has drawn interest from chemical companies such as Cipla, who have an interest in B12, and in NGOs such as the pernicious anaemia society who are keen to help the development of international patient groups who suffer with B12 deficiency as well as GAIN who are interested in the provision of B12 in developing countries.

All members of this team have strong records in the public understanding of science. All those involved in the project will engage in regular talks and outreach events through organized events by direct invitation, ensuring there is good dissemination with the general public on a range of important issues.

The skills acquired by those involved in this project include not only a wide range of important biological techniques ranging from microbiology and recombinant DNA technology through to process optimisation, food technology and food trials. The knowledge and techniques will provide those employed with skills that can be used across education and industry. The intellectual property resulting from this project will be discussed, shared and protected via the various Innovation and Enterprise Offices linked with the research programme. The research will be published in high impact journals and oral communications given at international conferences. Collectively, these approaches will allow the research to be brought to the attention of many other leading food and industrial companies.
Description we have developed and enhanced the extraction and purification of vitamin B12 from a strain of bacteria that we had previously engineered - with a longer-term view of using this strain to produce the vitamin as a supplement in India. Secondly, we are examining alternative foods as sources of this essential nutrient that would be suitable for a largely vegetarian population. Specifically, we are looking at whether Chlorella can be used as a suitable source of vitamin B12, by adding Chlorella to food bars. These bars have been developed and have been entered into a pilot study in order to determine whether the B12 within Chlorella is bioavailable.
Exploitation Route We have interest from several Indian companies about trying to develop B12 production in India. This would help address nutrition security within the country. We have also been in contact with several companies on the production of nutrition enhanced food bars that are rich in vitamin B12. The outcome of our trials will help determine whether this interest will be taken further. Measurable impacts of the research will therefore include the development of alternative B12 sources from algae, higher plants and fermented food. Patients and clinicians will benefit from greater access to information on B12 deficiency through the formation of interest groups that are associated with this study.
Sectors Chemicals,Education,Healthcare,Pharmaceuticals and Medical Biotechnology

Description Whilst our research is on-going, we have been in contact with several companies concerning the research that has so far been undertaken. There is strong interest in developing B12 production within India and with improved extraction procedures we are in a position to produce the nutrient at very competitive rates. Vitamin B12 is essential for red blood cell formation and for maintaining neural connections, so low levels of B12 cause problems both with certain types of anaemia and with cognition. Improving levels of B12 therefore addresses a key objective of this project and our initial pilot trials using algae-derived B12 to improve nutrition are currently underway prior to a longer term study. There is a lot of interest from a number of food bar manufacturers concerning the outcome of this trial. Further research into the interaction between bacteria that support algae is giving a better idea of the basis for this mutualism and this has implications for growing larger cultures in a more sustainable fashion. we have also started to look at the effect of B12 levels on the human gastrointestinal microbiome.
Sector Agriculture, Food and Drink,Chemicals,Education,Healthcare
Impact Types Economic

Description Set up CluB12
Geographic Reach Europe 
Policy Influence Type Contribution to new or improved professional practice
URL https://www.club-12.org
Description Innovation Hub for Improving Health and Nutrition through Biofortification (HERB Hub)
Amount £374,862 (GBP)
Funding ID BB/X010864/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 11/2022 
End 11/2027
Description Amrita Hazra 
Organisation IISER Pune
Country India 
Sector Public 
PI Contribution Developing improved methods for B12 production
Collaborator Contribution Have identified new ways in which components can be made
Impact Have identified some of the enzymes required to make components of the molecule.
Start Year 2019
Description Collaboration with Prof Arvind Lali and Dr Ann Anil 
Organisation Institute of Chemical Technology (ICT)
Country India 
Sector Academic/University 
PI Contribution The ICT researchers have developed a range of separation technologies, which are important to us for the purification of biologically produced commodities such as vitamin B12.
Collaborator Contribution Together we have applied for GCRF funding to help enhance the provision of the nutrient in India.
Impact We have successfully applied for GCRF funding
Start Year 2017
Description Collaboration with Prof Chittaranjan Yajnik 
Organisation KEM Hospital, Pune
Country India 
Sector Hospitals 
PI Contribution Prof Yajnik provides access to a number of patients who are severely deficient in vitamin B12 - and this has been the focus of a major GCRF application aimed at trying to enhance vitamin B12 levels in cultural foods.
Collaborator Contribution We have developed a scheme that will allow for the testing of a number of different vegetarian-type foods as vitamin-enriched sources of B12.
Impact We have been successful in gaining funding from a GCRF application, which aims to provide more B12 within India and has the long-term of reducing B12 deficiency within the country.
Start Year 2017