Proposal Form

STRATEGIC INDO-SWEDISH COOPERATIVE

PROGRAMME ON “HEALTH AND DISEASE PREVENTION”

A. PROJECT

1. Title of Project

Development of whey protein formulations for preventing metabolic syndrome

2. Specific research field

Field Sub-field

Innovative foods Sustainable foods for disease prevention

※ please be sure to be specific.

3. Project Duration

01/ 05/ 2017 ～ 30/ 04 / 2020 (3 years) (dd/mm/yyyy)(Years & Months)

※ Project duration should be 3 to 4 years all together.

2

4. Summary of Project

Keywords

Metabolic syndrome Type 2 diabetes obesity

Whey Dietary protein Product prototype

insulinogenic Clinical trials Metabolic risk markers

Objectives

To develop and evaluate functional food product prototypes for prevention of metabolic syndrome, manifested as obesity, type 2 diabetes and cardiovascular diseases globally, and also impacting socioeconomic development. The product will be based on dietary proteins from a plentiful resource formed as a by-product of food industry, with an ultimate aim to scale up and transfer the technology for commercial production targeting the Indian market.

Approach

The project involves a Swedish team comprising a university research group and a SME with expertise in protein chemistry and separation technology, and also on design of functional foods and product scale up, and an Indian team involving a R&D group with competence in dairy science and product development and a major producer of dairy products.

The project has been divided into 7 work packages out of which 4 are R&D related. The research will start with characterization of the locally produced protein containing by-product, especially with respect to the insulinogenic factors and also the physico-chemical properties relevant for food systems, followed by development of food prototypes particularly for Indian consumers. To obtain the proof of principle, the product prototypes will be tested on healthy subjects and those prone to metabolic syndrome in India in collaboration with a renowned hospital after relevant ethical permissions are obtained. The Swedish team will be in charge of the studies on characterization of the raw material and development of business model, while the Indian team will take the lead in product development for the Indian market. Both partners will be engaged or will have insight into all the work packages. The SME in the Swedish team will evaluate aspects concerning production scale up, product shelf life and distribution of the technology. The project will be coordinated by the Swedish partner, and will involve regular communication by different modes as well as exchange visits between the two countries. A consortium agreement will define the rights to any intellectual property arising from the cooperative project.

3

Expected

Outcome

Functional food product prototype(s), with possible patent protection

Novel functional foods with standardized protocol for manufacture

Data from the clinical trials

1-2 publications in international peer-reviewed journals

Eventually formation of a joint venture between the Swedish company and Indian distributor

※ Attention – font: Times New Roman, size: 11 points / Do not exceed space provided.

4

5. Personal Data of Principal Investigators

INDIA

Name of PI Sreeja V.

Title/ Designation Assistant Professor

Department Dairy Microbiology

Organization Anand Agricultural University

Address

Dairy Microbiology Department, SMC College of Dairy Science, Anand Agricultural University, Anand-388 110, Gujarat, India

Office phone #

Cell phone # +919998326542

Fax #

e-mail sreeja_p70@rediffmail.com, sreejamudgal@aau.in

Date of Birth 01/05/1971

SWEDEN

Name of PI Rajni Hatti Kaul

Title/ Designation Professor

Department Department of Chemistry

Organization Lund University

Address

Division of Biotechnology Center for Chemistry & Chemical Engineering Box 124, SE-221 00 Lund, Sweden

Office phone # 046-222 4840

Cell phone # 0708 748540

Fax # 046-222 4713

e-mail Rajni.Hatti-Kaul@biotek.lu.se

Date of Birth 04-07-1959

5

6. Project partners

Partners in Indian Team

Name Organization, Division Title Degree Specialty

Leader

Sreeja V.

Anand Agricultural University, SMC College of Dairy Science, Dairy Microbiology

Assistant Professor

PhD Dairy Microbiology

Partners

Jashbhai B. Prajapati

Anand Agricultural University, SMC College of Dairy Science, Dairy Microbiology

Professor PhD Dairy Microbiology

K. Rathnam AMUL dairy (Kaira District Co-operative Milk Producers Union Ltd)

Managing Director

PhD Nutrition

Subrota Hati Anand Agricultural University, SMC College of Dairy Science, Dairy Microbiology

Assistant Professor

PhD Dairy Microbiology

Partners in Swedish Team

Name Organization, Division Title Degree Specialty

Rajni Hatti Kaul Lund University, Division of Biotechnology Professor PhD

Biotechnology, Protein separation and characterization

Angeliki Triantafyllou

Mats Forsberg

Bo Mattiasson

Sub-contractor for Lund University

Sub-contractor for Lund University

Indienz AB

Senior scientist

Mr.

Professor Emeritus

Technology Dr.

Master of Science

PhD

Food processing and product development

Business development and product management

Biotechnology, bioseparations, bioanalysis

6

B. TECHNICAL INFORMATION

1. Objectives of Project (up to 200 words)

The aim of the project is to develop marketable functional food product prototypes from cheese whey, a renewable by-product of the dairy industry. The aim will be fulfilled through achievement of the following objectives:

Characterization of the locally produced whey and investigation of the effect of processing methods for whey concentrates/whey powders on their insulinogenic and physico-chemical properties

Development of product prototypes appealing to Indian consumers including a basic business model to guarantee the prototypes commercial viability.

Demonstration of the positive effect of the prototype(s) on metabolic risk markers through clinical trials

2. Justification for collaboration, including background to the proposed project, each partner’s expertise and specific contribution to the project, and the added value of the proposed collaboration (up to 400 words)

The idea underlying the project was initiated by the Swedish team based on the knowledge from the research performed at Lund university on the insulinogenic properties of whey protein and also motivated by the involvement of one of the team members (Dr. Angeliki Triantafyllou) in the Antidiabetic Food Center funded by Vinnova. The call for the Indo-Swedish collaborative research and innovation project presented a good opportunity for developing a product prototype of relevance for the growing numbers of adults and children afflicted with obesity, diabetes and cardiovascular diseases among the Indian population and the world at large. The Swedish team involves members with expertise in protein chemistry, separations and industrial applications, development of functional foods, business development and consultancy. The team has a large network of national and international contacts in academia and industry.

The team has had previous collaboration with the members of the Indian team at College of Dairy Science, Anand, concerning development of probiotics and biotechnological production processes. With a good knowledge of the dairy products and industry, experience in development of dairy based products, and also involvement of India´s largest producer of dairy products, Amul, makes the choice of the Indian partner obvious and highly justified.

The Swedish team will be responsible for project management and business development aspects, and also characterization of whey proteins, and will also be involved in the development of product prototype for which the Indian partner will be in charge. Clinical studies will be carried out primarily in India with the involvement of both partners.

7

3. Descriptions of the Cooperative research project – maximum 6 pages (font size 12 pt)

(Note: This should include expected outcome of the project incl. significance for industry and society, special strengths of the groups, the expected added value incl. how the groups complement each other, how cooperation can be strengthen over time, expected added value due to combination of different fields if applicable, worldwide comparison of similar activities, plan for exploitation of results, plan for contract between the parties and handling of Intellectual Property within the consortium.)

Background Impact Implementation Actor constellations Work plan (schedule, milestones and deliverables)

Development of whey protein formulations for preventing metabolic syndrome

1. Background

The main goal of this project is to develop formulation(s) based on whey protein for controlling the blood glucose level and the concomitant adverse effects in individuals at a risk for developing metabolic syndrome (MS), which can be manifested as obesity, cardiovascular disease and type 2 diabetes (diabetes mellitus), important causes of morbidity and mortality worldwide. India has been facing a rising burden of cardiovascular diseases over the past two decades, which is estimated to be the largest cause of disability and mortality in the country by 2020, with 2.6 million deaths. The prevalence of MS has been seen to be more prevalent in age group 41-60 years, obesity being one of the major causes for its development1.

Persons exhibiting metabolic risk factors related to MS benefit from lifestyle changes including dietary habits for controlling LDL cholesterol, fasting glycaemia, body weight and blood pressure. Earlier studies have shown that high protein diets lead to weight loss and greater satiety, certain proteins like milk serum proteins (whey proteins) being more effective than others and stimulate insulin secretion when co-ingested with carbohydrates2,3 .Besides regulating the appetite, the whey proteins have also high nutritional value because of the presence of all essential amino acids in higher concentrations than vegetable protein sources, and are particularly rich in branched chain amino acids with high leucine content that are more potent insulin secretagouges4. Whey proteins are easily digested and absorbed, resulting in a quick plasma-AA response, they have insulinogenic effect and reduce postprandial glycaemia5,6 . The insulinogenic effect of whey is attributed to the stimulating the release of several gut hormones resulting in slow gastric emptying7;8.

1 Sawant A, Mankeshwar R, Shah S, et al. (2011) Cholesterol doi:10.1155/2011/920983 2 Van Loon LJ, Saris WH, Verhagen H, Wagenmakers AJ (2000) Am J Clin Nutr 72, 96-105 3 Manders RJ, Wagenmaker AJ, Koopman R, etal (2005) Am J Clin Nutr 82, 76-83 4 Walzem RL, Dillard CJ, German JB (2002) Crit Rev Food Sci Nutr 42, 353-375 5 Nilsson M, Stenberg M, Frid AH, Holst JJ, Bjorck IM (2004) Am J Clin Nutr 80, 1246-1253 6 Gunnerud UJ, Östman EM, Björck IME (2013) Eur J Clin Nutr 67, 749-753 7 Jacubowicz D, Froy O (2013) J Nutr Biochem 24, 1-5 8 Mignone LE, Wu T, Horowitz M, Rayner, CK (2015) World J Diabetes 6, 1274-1284

8

India ranks first in milk production, accounting for 18.5% of world production; an annual output of 146.3 million tonnes was recorded during 2014-159. The recent trend in the production of indigenous milk products indicates a massive increase in the production of whey to over 3 million tonnes per year. Some of the applications include its use in butter milk, production of whey protein concentrate and for agricultural purposes but a considerable amount of whey is disposed as waste10. The large number of Indians suffering from diabetes, obesity and related cardiovascular diseases, would benefit from consuming health promoting foods rather than medicines with serious side-effects. This project has thus an ambition of designing and making available a whey based formulation with a desirable texture, stability and palatability, along with dose recommendations for subjects suffering from or prone to MS.

2. Impact This project fulfills a number of sustainable development goals regarding efficient use of a natural resource for promoting public health on one hand and reducing the environmental impact of disposal of the surplus resource on the other. India accounts for third highest level of obesity among both adults and children in the world in absolute terms. Type 2 diabetes is targeted as one of the priority non-communicable diseases for sustainable development; the number of adult diabetics in the world having increased almost four times from 108 million in 1980 to 422 million in 2014. Seventy seven percent of people with diabetics live in low- and middle-income countries. India, once known as the ‘diabetes capital of the world’ was home to over 61 million patients with T2D in 2011 with predictions of 101.2 million diabetics by 203011. The most disturbing trend is the shift in age of onset of diabetes to a younger age. Given the lifelong costs associated with diabetes, many individuals and families are unable to cope with the economic, emotional and social disease burden. Early identification of at-risk individuals and appropriate lifestyle interventions including dietary interventions would significantly help in preventing or postponing the onset of diabetes and related metabolic syndromes.

The commercial viability of the output from the project is vital for achieving the desired impact. We have therefore formulated a WP that defines and deliver the basic elements for the future development of a business plan after the project has ended. A basic business model with the project business logic will be delivered on technology readiness level 5.

3. Implementation

The project will involve two research partners, one in Sweden and one in India, together with one small company in Sweden and an important dairy cooperative in India. The principal investigators in Sweden and India will be responsible for managing the project on either side and to ensure that the research activities follow the work plan. They will also be responsible for maintaining good communication between the different partners and within their own organization. The PI in Sweden will also serve as a project coordinator. All the activities within the project are divided into a number of work packages, each with a work package leader responsible for putting together the

9 http://www.news18.com/news/business/economic-survey-2015-16-india-ranks-first-in-milk-production-accounting-for-18-5-per-cent-of-world-production-1208209.html

10 http://whey-production-scenario.blogspot.se/2012/06/scenario-of-whey-production-in-india.html

11 Gupta M et al. Int J Sci Rep. 2015 May;1(1):1-2

9

team and facilities required, and for delivering the results. Much of the communication between the Swedish and Indian partners will take place through e-mails and also through skype calls. Both sides will make research and consultation visits to each other´s laboratories at least once a year, starting with a kick-off meeting and final summing up meetings in India.

A consortium agreement will be drafted among the partners that will specify the formal modalities of collaboration and also the handling of intellectual property rights. This will be one of the important points of discussion during the kick-off meeting. In general terms, the ownership of IPR will depend on an individual/partner´s contribution to the results. The results from the project are to be made public in accordance with good international standards. But prior to publication the party(ies) should submit a copy of the planned publication to the other partners for review. A party interested in acquiring the results and seeking intellectual property rights protection can do so on commercially favorable terms and conditions. In such case, the publication may be delayed for a maximum of 90 days.

Indienz, the swedish SME collaborating in the project will focus on product scale up and also for defining key partnerships, cost structures, etc. for commercialization.

4. Actor constellation

The project partners have varied backgrounds and expertise that is complementary for fulfilling the aims of the project. Below is the description of the expertise of the Swedish and Indian teams:

Lund University (LU): The Biotechnology Division at LU is active in various research areas including industrial- and environmental biotechnology, bioseparations and bioanalysis. Prof. Rajni Hatti Kaul (RHK) has over 25 years of experience in research and development in biotechnology including protein purification and stabilization. She is currently leading two cross-disciplinary projects with strong industry involvement concerning valorization of agriculture residues and biobased plastics, respectively. LU will have two sub-contractors: Dr. Angeliki Triantafyllou (AT) for providing competence in food processing product development and quality assurance. AT has experience in prototype-to-market procedures for industrial manufacture of functional foods and she actively worked in the Vinnova sponsored Antidiabetic Food Center (AFC) for many years, on design and clinical trials. AT has also knowledge in cheesemaking. Mats Forsberg (MF) for developing a business model. LU will even involve Prof. Inger Björck as a consultant, who has been the director of Antidiabetic Food Center for 8 years and has studied the effect of whey proteins on postprandial glycaemia.

Indienz AB: is a biotech company in southern Sweden possessing lab to pilot scale facilities for microbial fermentations, product production and separation, biogas production, etc. The company serves the customers through contract research and development and also consultancy on biobased production, bioprocess design, etc. Prof. Bo Mattiasson has vast experience in different aspects of biotechnology for chemicals, biopharmaceuticals production and environmental remediation. He is also a founder of several companies.

Anand Agricultural University (AAU): Dairy Microbiology department, SMC College of Dairy Science at AAU in the state of Gujarat, India, conducts research related to Dairy Microbiology and Food Biotechnology as per the demand of the industry and policy of the university and participates in various extension activities, main research activity being exploitation of beneficial lactic acid bacteria for human health and development of products containing these probiotic bacteria. Department has also conducted work on development of rapid quality control tests, clean milk production, isolation and characterization of ACE inhibitory bioactive peptides from fermented milks, camel milk etc. Dr. Sreeja V., the PI of the Indian team specializes in fermented milks, Lactic acid bacteria, probiotics and their health benefits, probiotic and synbiotic products, dry probiotic formulations such as capsules, tablets. She has been working as Co-PI and PI in various projects

10

funded by Government of Gujarat, ICAR, DBT and GSBTM. Dr. JB Prajapati, an expert in the area of synbiotic product develiopment and its nutritional and therapeutic aspects will associate with the project as Co-PI. Another Co-PI of the project is Dr. Subrota Hati, whose area of specialization also includes ACE inhibitory peptides.

AMUL dairy: Kaira District Co-operative Milk producer’s Union, popularly known as AMUL, is an Indian dairy cooperative formed in 1946 and is based at Anand. It manufactures a number of dairy products under the AMUL brand name. It will be involved as a formal partner in the project. Dr. K Rathnam, Managing director, AMUL dairy has vast experience in the various aspects of dairy field.

Work Plan – Schedule, milestones and deliverables

The work plan is envisaged over a period of 36 months and is structured in 7 work packages (WPs) of which one WP relates to project initiation and management (WP1), 4 WPs relate to RTD activities (WPs2-4 and 6), and 2 WPs relate to future commercialisation and dissemination of research outputs (WP 5 and 7). The duration and timing of each work package is illustrated in Table 1.

Table 1 - Gantt chart indicating time duration for each of the work packages (WPs 1-7)

MONTHS 1-6 7-12 13-18 19-24 25-30 31 -36

WP1 (M1-36)

WP2 (M1-8)

WP3 (M1-12)

WP4 (6-24)

WP5 (M13-30)

WP6 (M13-30)

WP7 (M1-36)

Work package 1 - Project initiation, Project management and monitoring (Lead partner: LU)

Objectives: Successful implementation of the project

Description of work: This WP aims to ensure the successful implementation of the project within the proposed budget and timeframe and smooth collaboration between the partners.Task-­­1. Overall project management. The project co-ordinator will chair the project Management Board (MB) which will also consist of the leading investigators from each partner Institution/Company. The MB meetings will be held regularly on a quarterly basis via videoconference (e.g. Skype). . Task-­­2. Financial monitoring in relation to the objectives of the project. Financial monitoring and administration support will be managed and provided by MF that reports to RHK.

Deliverables: D1.1 Project Management Board (MB) reports (Month 12 & Month 24). D1.2 Written final report (Month 36)

Work package 2 - Characterization of whey by-product from cheese production (Lead partner: LU)

11

Objectives: Characterization of whey produced using different methods i.e. enzymatic and fermentation as well as acid treatments of milk from locally farmed animals to assess its nutritional quality

Description of work: Treatment of milk using the enzyme rennet is the routine method of producing cheese in Europe, while in India cheese is commonly produced by directly lowering the pH by addition of vinegar or lime juice. Characterization of whey obtained is of primary importance in order to assess its nutritional quality. The work will be initiated in Sweden and will involve a person from the partner institution in India, who will apply the analytical methods on the whey product from India.

Task n.1. Obtaining milk and whey from local sources. Milk and whey from the local diaries such as Skånemejeriet in Southern Sweden and Amul in Anand, India will be obtained and stored in liquid form or as freeze-dried powder (LU, AAU, Amul) Task n.2. Determination of protein content and amino acid composition. Analysis of the different whey preparations will be done using standard methods. The content and proportion of insulinogenic aminoacids will be determined. Moreover the effect of different process operations such as protein fractionation, concentration and drying procedures on the composition of the whey product will be studied. In that way material losses and potentially decreased nutritional value due to for example heat induced Maillard reactions will be documented. (LU, AAU)

Deliverables and month of delivery: D2.1. Reports on the composition of insulinogenic amino acids in Swedish- and Indian whey products (Month 8)

Work package 3 – Determination of physical and technological properties for food development (Lead partner: LU)

Objectives: To determine functional and technological properties e.g. gelation behavior, emulsification capacity and gel strength, of the whey preparations used in Task 1

Description of work: Task n.1. Determination of rheology of the whey preparations. Rheology will be determined under various conditions that are relevant for the food systems in terms of pH, ionic strength and temperature. (LU) Task n.2. Behaviour of the whey preparations in emulsions. Assessment of taste, texture and mouthfeel, homogeneity and stability of foods and beverages that will be developed. These food prototypes could be high-protein, viscous lassi, tea made with the whey-rich preparation instead of traditional tea and milk, and whey enriched traditional breads such as dosa and new co-extruded breakfast cereal products. (LU) Task n.3. Investigate the effect of process operations on model products characteristics including QA aspects

Deliverables and month of delivery: D3.1. Report on rheology and behaviour of the whey preparations in emulsions (M14) D3.2. Draft manufacturing lines for various types of products

Work package 4 – Development of food product prototypes (Lead partner: AAU)

Objectives: Development of whey based product prototypes to facilitate their incorporation in peoples´ diet considering the eating patterns and preferences of various consumer groups (gender, age, occupation/ lunch habits), in order to facilitate substitute foods already consumed.

Description of work: The amount of whey protein that is thought of providing blood glucose regulating effect according to other studies will be incorporated in various food items for breakfast, lunch, snacks and dinner.

12

Task n.1 Adjustment and optimization of the production protocol (AAU, Indienz, LU) Task n.2 Sensory, textural and shelf stability, and consumer acceptability of the product prototypes Task n.3 Supply of test produts for the clinical trials (AAU, Indienz)

Deliverables and month of delivery: Product prototypes with determined characteristics available (M24)

Work package 5 – Developing a business model for refined whey protein (Lead partner: LU)

Objectives: To develop a business model for refined whey protein. We will develop the rationale of how we will create, deliver and capture value for the partners in the consortium and the society. This WP will guarantee the commercial viability of the output of the project.

Description of work: Based on Business Model Generation by Osterwalder.

Task – 1: Defining customer segments. Task – 2: Defining the value propositions. Task – 3: Defining the channels. Task – 4: Defining the customer realtionships. Task – 5: Defining the revenue streams. Task – 6: Defining the key resources. Task – 7: Defining key activities. Task – 8: Defining key partnerships. Task – 9: Defining the cost structure.

Deliverables and month of delivery: A business model (TRL 5) for refined whey protein. This process of business model construction will be a part of a future business strategy for transferring the developed technology at an industrial scale level in order to launch the commercial production of the new products. (M30)

Work package 6 – Clinical trials of the effects of the product prototypes (Lead: AAU)

Objectives: To study the effect of the product prototypes from WP4 on metabolic risk factors in healthy subjects and those prone to metabolic syndrome

Description of work: Double blind, randomized cross-over design will be used to study the effect of the whey preparations as compared to a traditional diet on healthy adults, males and females in the age group 40-60, Each diet phase will last 4 weeks with a 4 week washout period. The clinical trials will be done in collaboration with a hospital in India, and appropriate ethical permissions will be obtained prior to the study. Individuals that comply with the inclusion criteria will be provided test meals for a period of at least 6 weeks. The volunteers will be advised regarding their food intake. A food diary will be registered. An accute study on glucose metabolism will be carried in the beginning of the intervention where the test product will be compared with a standard meal for breakfast, following a controlled evening meal. Moreover a long-term study will be performed in which in addition to blood glucose and insulin, also lipids and inflammation markers (e.g. IL-6, TNF-a) will be measured. In the end of the intervention body weight, lean muscle mass and bone density will be determined.

Task n.1. Blood analysis. Gut hormones (e.g. PYY, GLP1) that indicate appetite control, as well as blood glucose and insulin values at intervals upto 4 hours after intake. (LU, AAU) Task n.2. Determination of long term metabolic effects and lean mass and bone density. Body scan DEXA will be performed at the beginning and at the end of the study to determine potential loss of fat and increase of lean mass as there is great support from the literature regarding the positive effect of high protein intake for maintaining muscle and increased bone density (particularly for the elderly) and for increasing lean at the expense of fat tissue after exercise. (LU, AAU)

13

Deliverables and month of delivery: D6.1 Compiled results of the clinical trials (M32) D6.2 Consumer acceptability assessment

Work package 7 - Dissemination of research outputs and intellectual property, knowledge protection and regulatory issues (Lead partners: LU, AAU)

Objectives: To determine patentability of the product prototype(s), dissemination of the research outputs, and establish contact with relevant stakeholders

Description of work: Knowledge protection: It is likely that the work in this project will allow a patent application. This would be assigned to Indienz AB, whereas fees to all the participants will be regulated. When appropriate, for the far-away markets, the technology may be licensed to a suitable local partner or customer to ensure its controlled production with all related benefits secured for the company. A web site (a web page on the existing web site of Lund university) will be developed and maintained in order to provide consumers awareness of the health and nutritional benefits of the innovative whey protein formulations for preventing the metabolic syndrome (without compromising release of sensitive information linked to intellectual property). Publications will be in the format of research manuscripts submitted to academic journals and conferences (after addressing intellectual property interests across the consortium). Project advances will also be extended to industry particularly through attendance at ‘food & drink and healthcare fair’ events as well as other innovation relay events within Europe and India. In addition, the input provided by the industrial end-user partner will play an important role. They will advise the consortium at appropriate phases of the project on the most interesting targets for their applications and thus safeguard a close feeling and contact with the market demands.

Deliverables and month of delivery: D7.1 Publications in peer review journals and/or participation in international conferences. (M24-36). D7.2 Development of a web site for the consumer’s awareness. (M24) D7.3 Expected patent publication, potentially trademark registration

14

Form-5

4. Plan for the execution of the Cooperative Project

The work plan of the project is divided into 7 work packages, four of which involve research activities.

The research activities that will primarily be carried out in Sweden, also with involvement of Indian partners are the following:

Characterization of the whey by-product obtained by different methods

The characterization of whey that is produced commercially by different methods (enzyme/fermentation or acid) and from locally farmed animals is of primary importance in order to assess its nutritional quality. The content and proportion of insulinogenic aminoacids in the whey preparation will be determined. Moreover the effect of different process operations such as purification, concentration and drying procedures on the composition of the whey product will be studied. In that way material losses and potentially decreased nutritional value due to for example heat induced Maillard reactions will be documented.

Determination of physical and technological properties for food development

The composition of whey and its method of manufacture will affect its functional and technological properties e.g. gelation behavior, emulsification capacity and gel strength. Rheology of the whey preparations will be determined under various conditions that are relevant for the food systems in terms of pH, ionic strength and temperature. Also the behavior of the whey preparations in emulsions will be studied. The taste, texture and mouthfeel, homogeneity and stability of foods and beverages prototypes will be studied. These food prototypes could be high-protein, viscous lassi, it could be tea made with the whey-rich preparation instead of traditional tea and milk and whey enriched traditional breads such as dosa and new co-extruded breakfast cereal products.

The major activities in India will be:

Development of food product prototypes

Product prototypes will be developed to facilitate the incorporation of this healthy component in peoples´ diet considering the eating patterns and preferences of various consumer groups (gender, age, occupation/ lunch habits). The foods would easily substitute those already consumed. The amount of whey protein that is thought of providing blood glucose regulating effect according to other studies will be incorporated in various food items for breakfast, lunch, snacks and dinner.

Clinical trials of the effects on healthy adults

Individuals that comply with the inclusion criteria will be provided test meals for a period of at least 6 weeks. An acute study (cross-over, randomized, double blind) on glucose metabolism will

15

be carried in the beginning of the intervention where the test product will be compared with a standard meal for breakfast, following a controlled evening meal. Gut hormones (e.g. PYY, GLP1) that indicate appetite control, as well as blood glucose and insulin values at intervals upto 4 hours after intake will be determine. Moreover a long-term study will be performed where in addition to blood glucose and insulin, also lipids and inflammation markers (e.g. IL-6, TNF-a) will be measured. In the end of the intervention body weight, lean muscle mass and bone density will be determined to study the effect of extra whey protein intake on this tissue. Cooperation and exchange

The above activities will involve cooperation between both Swedish and Indian teams. Regular contact will be maintained through emails and skype meetings, which will be coordinated by the Swedish PI. There will also be physical exchanges between the countries. The living costs of the visiting researchers will be covered by the host institution.

We plan to start the project with a kick-off meeting in India where all the PIs that have been involved in drafting the proposal will participate. In the meeting, the plan for the entire project period, especially the consortium agreement and milestones for the first year, and any imminent risks will be discussed. Ownership to any IPR arising from the project will be defined by the consortium agreement. We will also take the opportunity to visit the Amul dairy to obtain first hand information on production volumes and present outlets for whey.

During year 2, we plan to have a workshop in Anand with focus on functional foods that will be open for both academia and industry. This will give us the opportunity of presenting our findings from the project.

Finally, at the end of year 3, there will be summing up meeting where the results from clinical trials will be available and discussed, and the way forward is planned. Publications and eventual patent applications will be planned. We will also plan meetings with potential stakeholders identified during the earlier years.

The research in Sweden will be done not only on the whey obtained from the Swedish dairy but also from India. One of the team members from India will visit Lund during first two years to learn the techniques of protein characterization and separation that will be applied on the raw material from India.

For the development of product prototypes and clinical trials in India, the Swedish team members especially Angeliki Triantafyllou will be involved in planning and monitoring the work. We have already established contact with a highly renowned hospital (P.D. Hinduja Hospital) in Mumbai, India where investigations on the prevalence of metabolic syndrome have been done (see reference 1 on page 7). The Department of Laboratory Medicine at the hospital is willing to help us with the clinical trials with our product prototypes, and will also take care of the ethical permissions. The Swedsih PI will have a meeting with the Head of the group in January 2017 in Mumbai.

16

5. Facilities related to project activity available at the institutions where the project will be carried out:

At the Collaborating Indian Institutions At the Collaborating Swedish Institutions

Facilities for product development and analysis such as pilot dairy plant facility for various dairy products, HPLC, fermentor, spectrophotometers, textural analyzer, gel electrophoresis, RT PCR, Sonicator, facilities for bacterial culture handing, preservation and production of fermented foods and other foods.

Facilities for characterization, analysis and separation of proteins like: chromatography, spectrophotometers, mass spectrometer, gel electrophoresis, membrane filtration up to pilot scale, and access to equipment for hysico-chemical characterization like determining rheology, emulsion behaviour, etc.

6. Information of visiting persons for technical mission (per annum).

Estimation of the expected plan of visits; any further modification, according to the needs of joint project has to be communicated and approved by the participants institutions.

A. India to Sweden

Year Name Organization Duration Purpose

1st Year PI AAU 2 weeks Visit to Lund university lab facility, visit to Indienz AB

1st Year Research

Fellow AAU 2 months

Preparation, study and analysis of different whey samples

2nd Year Co-PI AAU 2 weeks Interim review meeting

3rd Year PI/Co-PI AAU 2 weeks Final meeting and future prospects

B. Sweden to India

Name Organization Duration Purpose

1st Year

RHK

AT

MF

BM

LU

LU

LU

Indienz

30 days

5 days

5 days

5 days

Kick-off meeting, visit to the dairy company, project start

17

2nd Year

RHK

Res ass.

AT

MF

BM

LU

LU

LU

LU

Indienz

15 days

30 days

5 days and 15 days

5 days

5 days

Workshop,

Product development activities

Planning of clinical trials

Business model discussion

Stakeholder contact

3rd Year

RHK

AT

MF

BM

LU

LU

LU

Indienz

30 days

15 days

5 days

5 days

Final meeting, future planning, monitoring clinical trials

Business model discussions

Meeting with stakeholders

※ Attention: Please add lines if necessary.

Abbreviations: RHK, Rajni Hatti-Kaul; AT, Angeliki Triantafyllou; MF, Mats Forsberg; BM, Bo Mattiasson; Res ass., Research assistant

7. Expected results of Cooperation (e.g. joint publications, patents etc.)

Are any of the expected results likely to have commercial value? (up to 100 words)

The cooperation project is expected to result in a potential trademark registration, 1-2 joint publications and 1patented product prototype for scale up and demonstration. The prototype will be developed as a niche branded product with good commercial value, considering the growing awareness of foods and food habits and also the need for easy, readymade solutions for a healthy lifestyle.

18

8. Personal and Professional Data (CV) of Indian and Swedish PIs must be attached.

(CV must describe the expertise of the PIs in the proposed field of work by citing relevant scientific publications or patent applications during the last 5 years. Do not exceed two pages A4 size for each PI (Font: Times New Roman, size: 11 points).

See attached files

9. Complementary resources.

Cite other national or international funding resources for related scientific projects including funding agencies, funding period and grant amount.

Sr No.

Title of Project Funding Agency

Amount (Rs lakhs)

Date of sanction and Duration

Particulars of work done

1 Development of dairy starter cultures and value added dairy products

Govt. of Gujarat

15.2 2014- continued

Working as Co-PI.

2 NRS on Equines including Veterinary Type Culture – Multi-centered

ICAR 11.5 2009- continued

Working as Co-CCPI

3 Metagenomic and Clinical investigation of synbiotic fermented dairy product containing probiotic Lactobacillus helveticus MTCC 5463 in geriatric volunteers.

DBT 48.9lakhs Completed on 18-04-2015

Worked as Co-PI.

4 Bioprospecting of oxalate degrading lactic acid bacteria to develop a functional product with potential in preventing kidney stone disease

Gujarat State Biotechnology Mission (GSBTM)

27 lakhs 2016-2019 Working as PI

19

C. FINANCIAL INFORMATION

Budget or resources for the project; one for the Swedish side and one for the Indian side

Indian side

Sl# Item Year I Year II Year III Total

Indian Rupees

A. Non-Recurring

Equipments

Fermentor (fully automatic, 20 li capacity)

15,00,000 - - 15,00,000

Freeze dryer 35,00,000 - - 35,00,000

Total 50,00,000

B. Recurring

1. Human Resource

JRF/SRF -1 3, 36,000 3, 36,000 3, 36,000 10,08,000

Technical Assistants-1 96,000 96,000 96,000 2,88,000

Total 4,32,000 4,32,000 4,32,000 12,96,000

2. Consumables

Raw materials, chemicals, glasswares, reagents, kits, animals, etc.

10,00,000 10,00,000 10,00,000 30,00,000

3. Travel

International 500,000 500,000 500,000 15,00,000

Domestic 1,00,000 1,00,000 1,00,000 300,000

4. Contingency-Misc, hosting of Swedish partners, etc.

200,000 200,000 200,000 6,00,000

Clinical trial in collaboration with a hospital

- - 20, 00,000 20, 00,000

TOTAL 72,32,000 22,32,000 42,32,000 1,36,96000

5. Overheads (10% of above) 7,23,200 2,23,200 4,23,200 13,69,600

Total

79,55,200 24,55,200 46,55,200 1,50,65,600

20

Swedish side

- 1st year Costs LU (SEK) Indienz (SEK)

Personnel 510 256 243 500

Sub-contracting 128 000 -

Consumables 30 000 -

Analyses 10 000 -

Travel 30 000 10 000

Living costs for Indian partners in Lund

54 000 -

Total direct costs 762 256 253 500

Indirect costs 533 579 76 050

Total Year 1 1 295 835 329 550 Vinnova: 230 685, Cofinancing: 98 865

Total Year 1, applied from Vinnova

1 526 520 SEK

- 2nd year Costs LU (SEK) Indienz (SEK) Personnel 523 012 359 000 Sub-contracting 384 000 - Consumables 20 000 - Analyses - Travel 50 000 10 000 Living costs for Indian partners in Lund

27 000 -

Total direct costs 1 004 012 369 000 Indirect costs 702 808 110 700 Total Year 2 1 706 820 479 700

Vinnova: 335 790 Cofinancing: 143 910 Total Year 2, applied from Vinnova

2 042 610 SEK

- 3rd year Costs LU (SEK) Indienz (SEK)

Personnel 186 988 243 500

Sub-contracting 256 000 -

Consumables - -

Analyses - -

Travel 30 000 10 000

Living costs for Indian partners in Lund

27 000 -

21

22

23

Principal Investigator (Indian side)

1. Name : Dr. SREEJA V.

2.Designation: Assistant Professor

3.Complete Postal Address, Telephone Number, Fax, e-mail etc.

Department of Dairy Microbiology, SMC College of Dairy Science, Anand Agricultural University, Anand, Gujarat-388 110. Telephone: 02692-225852 Fax: 02692-261314 sreeja_p70@rediffmail.com, sreejamudgal@aau.in 4.Date of Birth: 1st May 1971

5. Educational Qualification : Degrees obtained (Begin with Bachelor’s Degree)

Sr No. Institution

Place

Degree

Awarded

Year Field of Study

1 Gujarat Agricultural University

M. Sc (Dairying) 1996 Dairy Microbiology

2 Anand Agricultural University

Ph.D (Dairy Microbiology)

2014 Dairy Microbiology

6. Research/Training Experience: Ongoing Research Projects

Sr No.

Title of Project Funding Agency Particulars of work done

1 Development of dairy starter cultures and value added dairy products

Gov. of Gujarat Working as Co-PI.

Isolation, characterization and deposition of lactic cultures, development of functional fermented milk products, development of probiotic dosage forms

2 NRS on Equines including Veterinary Type Culture – Multi-centered

ICAR Working as CCPI

Isolation, characterization and deposition of lactic cultures from traditional fermented foods of western states of India for biodiversity preservation, Studying the isolates for their functional attributes and probiotic characteristics

3 Bioprospecting of oxalate degrading lactic acid bacteria to develop a functional product with potential in preventing kidney stone disease

Gujarat State Biotechnology Mission (GSBTM)

Working as PI

Isolation, characterization and deposition of oxalate degrading lactic cultures, their application in product development, Functional evaluation of the product for potential application for kidney disease prevention

Completed Research Projects

Sr No.

Title of Project Funding Agency

Particulars of work done

24

1 Metagenomic and Clinical investigation of synbiotic fermented dairy product containing probiotic Lactobacillus helveticus MTCC 5463 in geriatric volunteers.

DBT Worked as Co-PI.

Developed synbiotic dairy products, Involved in the feeding trial and analysis aspects.

Trainings:

1) ICAR sponsored III short course on ”Metagenomics: Role of next generation sequencing and bioinformatics” held at department of Animal Biotechnology, College of Veterinary Science and Animal Husbandry, AAU, Anand during 6-15, October 2014.

2) Three months International training in the area of ‘Fermentation Technology’ under NAIP of ICAR from July 1st 2011 to September 30th 2011 at Lund University, Lund, Sweden.

3) Refresher course for faculty on “Value added Probiotic, Prebiotic and Synbiotic Products” held at SMC College of Dairy Science under the aegis of ICAR Niche area of Excellence for 21 days during 15 November to 5th December 2010.

4) Winter school on ‘Recent Advances in dairy neutraceuticals and Bioinformatics Applications” sponsored by ICAR, New delhi, held at NDRI, karnal for 21 days from 1-21 February, 2010.

7.Research specialization (Major scientific fields of interest)

Probiotics and health benefits, Lactic acid bacteria, Fermented foods

8.Important recent publications (last 5 years, with titles and References), including papers In press

1. V. Sreeja, Jashbhai Prajapati, Vaishali Thakkar , Tejal Gandhi and Vijaykumar Darji (2016). Effect of Excipients on Disintegration, Viability and Activity of Fast Disintegrating Tablets Containing Probiotic and Starter Cultures. Current Trends in Biotechnology and Pharmacy. 10 (2) 108-117, ISSN 0973-8916 (Print), 2230-7303 (Online)

2. Shruti Sharma, Sreeja V and Jashbhai B Prajapati (2016). Development of synbiotic lassi containing honey: Studies on probiotic viability, product characteristics and shelf life. Indian J Dairy Sci 69(2): 148-153.

3. Gohel, MK, Prajapati JB, Mudgal Sreeja V, Pandya HV, Singh, UV, Trivedi SS, Phatak,AG and Patel RM (2016). Effect of Probiotic Dietary Intervention on Calcium and Haematological Parameters in Geriatrics. Journal of Clinical and Diagnostic Research. 10(4): LC05-LC09.

4. Sreeja V. and Jashbhai B. Prajapati (2016). Probiotics for the geriatric population. In: Probiotics- from bench to community, edited by BS Ramakrushna, G Balakrish Nair and Yoshifumi Takeda, Elsevier RELX India Pvt Ltd, Gurgaon, pp 94-111. (a publication of Gut Microbiota and Probiotic Science Foundation).

5. V. Sreeja and Jashbhai B. Prajapati. (2013). Probiotic Formulations: Application and Status as Pharmaceuticals—A Review. Probiotics & Antimicro. Prot. DOI 10.1007/s12602-013-9126-2. 5 (2): 81-91.

6. Sreeja V, Atanu Jana, Kishore Aparnathi and Jashbhai Prajapati (2013). Role of whey proteins in combating geriatric disorders. J Sci Food Agric . 93: 3662–3669.

7. Senan S, Prajapati J, Joshi C, Sreeja V, Gohel M, Trivedi S, Patel R, Pandya H, Singh U, Phatak A and Patel H (2015) Geriatric Respondents and Non-Respondents To Probiotic Intervention Can Be Differentiated By Inherent Gut Microbiome Composition. Front. Nutr.2:25. doi:10.3389/fnut.2015.00025.

8. Sreeja V. and Prajapati JB (2015). Bifidus milks. In: Fermented milk and dairy products, edited by A.K. Puniya, CRC Press- Taylor and Francis Group, BotaRacon, pp. 269-310. ISBN: 978-1-4665-7797-8, eBook ISBN: 978-1-4665-7800-5.

25

Commissions of trust

26

Programme Director of Mistra programme, STEPS, 2016-2020 Coordinator of Formas project, FARM2FURAN, 2016-2020 Head of the Division of Biotechnology, Lund University, 2006-2014 Program Director of Mistra sponsored research programme, Greenchem (Specialty chemicals from

renewable resources), 2003-2010 Scientific Coordinator of Vinnova sponsored research project, BioVinn (Industrial biotechnology for

production of platform chemicals), 2008-2013 Scientific Advisory Board member, Toulouse White Biotechnology, Toulouse, France, 2014-to date Reviewer of research proposals to Austrian Science Fund; ERA-IB; Danish Council for Strategic

Research; Cariplo Foundation, Italy; EUs Innovative Medicines Initiative; Agence Nationale de la Recherche, France, Science Foundation Ireland

Editorial positions

Review Editor, Frontiers in Bioengineering, 2015- Senior Editor, Cogent Biology, 2015- Editor, Biotechnology Reports, 2014- Editorial Board member, Molecular Biotechnology, 2000-2007

Faculty Opponent/Examiner at PhD defences Member of examination committees for PhD defences in Sweden, >20 times External examiner for 2 PhD defences in Denmark, and several PhD and -Master theses from several countries including India, Sri Lanka, Malaysia, Kenya Networking activities Co-organised 3 international conferences and 1 symposium related to industrial biotechnology in Sweden, and 1 international conference and 1 workshop in India Selected publications from past 5 years Dishisha T, Ståhl Å, Lundmark S, Hatti-Kaul R (2013) An economical biorefinery process for propionic acid production from

glycerol and potato juice using high cell density fermentation. Bioresour. Technol. 135, 504-512 Van-Thuoc D, Hashim SO, Hatti-Kaul R, Mamo G (2013) Ectoine mediated protection of enzyme from the effect of pH and

temperature stress: a study using Bacillus halodurans xylanase as a model. Appl. Microbiol. Biotechnol. 97, 6271-6278 Irani M, Törnvall U, Genheden S, Wittrup Larsen M, Hatti-Kaul R, Ryde U (2013) Amino acid oxidation of Candida antarctica

lipase B studied by molecular dynamics simulations and site-directed mutagenesis. Biochemistry 52, 1280-1289 Sabet-Azad R, Linares-Pastén JA, Torkelson L, Sardari RRR, Hatti-Kaul R (2013) Coenzyme A-acylating propionaldehyde

dehydrogenase (PduP) from Lactobacillus reuteri: Kinetic characterization and molecular modeling. Enzyme Microb. Technol.53, 235-242

Gaber Y, Ismail M, Bisagni S, Takwa M, Hatti-Kaul R (2015) Rational mutagenesis of pig liver esterase (PLE-1) to resolve racemic clopidogrel. J. Mol. Catal. B: Enzymatic 122, 156-162

Villagomez V, Hatti-Kaul R, Sterner O, Almanza G, Linares-Pastén JA (2015) Effect of natural and semisynthetic pseudoguianolides on the stability of NF-κB:DNA complex studied by agarose gel electrophoresis. PLOS ONE 10(1): e0115819.doi:10.1371/journal.pone.0115819

Abouhmad A, Mamo G, Dishisha T, Amin MA, Hatti-Kaul R (2016) T4 lysozyme fused with cellulose binding module for antimicrobial cellulosic wound dressing materials. J. Appl. Microbiol.121, 115-125

Alkhalili R, Bernfur K, Dishisha T, Mamo G, Schelin J, Canbäck B, Emanuelsson C, Hatti-Kaul R (2016) Antimicrobial protein candidates from the thermophilic Geobacillus sp. strain ZGt-1: production, proteomics and bioinformatics analysis. Inter. J. Mol. Sci. 17, 1363

Rahimpour F, Hatti-Kaul R, Mamo G (2016) Response surface methodology and artificial neural network modeling of an aqueous two-phase system for purification of a recombinant alkaline active xylanase. Process Biochem. 51, 452-462

27