Whey lactose bioconversion in hight-added value products using cheap renevable substrate , OLIGOLAC

ABOUT THE PROJECT

  • Eureka number: E!8076
  • Finance contract number: 335E/19.12.2013
  • Project title: Whey lactose bioconversion in high-added value products using cheap renewable substrate
  •  Acronym: OLIGOLAC
  • Project coordinator: ROMVAC COMPANY S.A. –General Manager Eng. Constantin Chiurciu
  • Project manager : Ion Nicolae
  • Financed by: The Executive Unit for Financing of Higher Education, Research, Development and Innovation (UEFISCDI) –INOVARE Program –Eureka-Eurostars European Cooperation Subprogram
  • Link to project page on Eureka website: http://www.eurekanetwork.org/project/id/8076
  • Link to UEFISCDI page: www.uefiscdi.gov.ro
  • Total contract value: 2.819.500 lei
  • Contract value (state budget only): 1.290.000 lei
  • Duration of implementation:  19.12.2013-15.11.2016

SYNDICATE TEAM

  • Project coordinator
    Romvac Company S.A.
    Sos. Centurii, nr. 7, Ilfov, 077190, Voluntari
    Project manager – Dr. Ion NICOLAE; romvac74@gmail.com
  • Partner P1
    The Research-Development Institute for Chemistry and Petrochemistry  – ICECHIM Bucharest
    Spl. Independentei 202, sector 6, 06021, Bucharest
    Project responsible –Biologist Dr. Diana PASARIN; dianapasarin11@yahoo.com
  • Partner P2
    The National Research-Development Institute for Food Bioresources
    Str. Dinu Vintila nr. 6, sector 2, Bucharest
    Project responsible – Biologist Dr. Lavinia BERCA; laviniamariana.berca@gmail.com
  • Foreign partner – SLOVENIA
    Emona – Nutrion Research & Development Department
    Kavciceva, 72, 1000 – Ljubljana, Slovenia
    Project responsible – Dr. Sc. Matjaz Cervek; matjaz.cervek@e-rcp.si

OBIECTIVE

Production of monocellular protein, oligosaccharides known as Bifidus growth factor and chelated amino acids, using highly-productive lactic yeasts grown on cheap substrate, namely whey, by changing the biotechnological parameters

ANTICIPATED PROJECT RESULTS

  • Obtaining high-performance lactic yeast strains
  • New feed additives: monocellular protein, oligosaccharides and chelated amino acids
  • Optimum feed mixtures in which the protein component is replaced with the mentioned feed additives

STEPS

Step I – Assessment of starting material (whey) and production of various efficient and cost-effective fermenting media. Preparation of growth conditions and monitoring of animal trial groups. Maximization of technological parameters for selection of new adapted and highly-productive lactic yeast strains and their preservation, Period: 19.12.2013 – 15.11.2014

Step II – Production and description of bioproducts. Elaboration of the technological laboratory process and experimental production, Period: 16.11.2014 – 15.11.2015

Step III – Demonstration and assessment of technological efficacy, Period: 16.11.2015 – 15.11.2016

Step IV – Communication of CO-ROMVAC results , Period: 16.11.2016 – 15.11.2017

Step V – Communication of CO-ROMVAC results , Period: 16.11.2017 – 15.11.2018

Step VI – Communication of CO-ROMVAC results , Period: 16.11.2018 – 15.11.2019

PHASED ACTIVITIES

Phase I – Assessment of starting material (whey) and production of various efficient and cost-effective fermenting media. Preparation of growth conditions and monitoring of animal trial groups. Maximization of technological parameters for selection of new adapted and highly-productive lactic yeast strains and their preservation

  • Modernization of SPF poultry breeding and exploitation farm, loft improvement for pigeon trials. Selection and organization of trial and control animal groups. Training of staff for application of method
  • Physicochemical assay of the energetic substrate (whey) by Munson-Walker and ICP-OES methods. Determination of composition of culture media (growth media)

      

  • Microbiological description of starting materials as per dairy products standards
  • Obtaining media for filling and preserving by freeze-drying or nitrogen freezing of adapted lactic yeast strains

   

  • Lab trials for obtaining adapted lactic yeasts by changing technological parameters (temperature, pH, ventilation, inoculums size) starting from parent strains (Kluyveromyces marxianus, K. marxianus marxianus)

   

Colonii de Kluyveromyces marxianus

    • Phenotypic and genetic description of parent lactic yeast strains
    • Freeze-drying or liquid nitrogen immersion of adapted lactic yeast strains; determination of stability of products by accelerated and real-time degradation assays
    • Selection of new adapted lactic yeast strains, depending on the starting material (whey) conversion yield into protein biomass (yeast)

    Phase I  Results         –          

Phase II – Obtaining and description of bioproducts. Elaboration of the technological laboratory process and experimental production

  • Morphophysiological, cultural and molecular description of new adapted and selected yeast strains

Aspectul coloniilor Kluyveromyces               Kluyveromyces marxianus marxianus                                                  

    marxianus pe Sabouraud                                          Frotiu Coloratie Gram

  • Obtaining feed supplements (additives) with lactic yeasts as such or combined with other pre/probiotics and excipients, in the lab

OLIGOLAC – ANIMAL PREMIX, bulk product packed into aluminum bags

  • Obtaining and physicochemical and microbiological description of yeast biomass. Isolation and analytical description of oligosaccharides by SM and RMN techniques

Gram staining determination of yeast cell viability which were lysed by autolysis, ultrasound and bile mill + autolysis

  • Experimental model for obtaining microelements chelated with amino acids and their description in the solution and in the dried product by HPLC and ICP-OES

Spectrometru GF-AAS AAnalyst 600                              Spectrometru ICP-MS NexION 300Q

  • Recording, storage and analysis of data necessary for the industrial development of pilot trial for determination of optimum technological parameters

        

Freeze-drying station

Phase II results              –  

Phase III – Demonstration and assessment of technological efficacy

Phase III results              –  

  • Lab technology for obtaining yeast biomass and oligosaccharides

                                               

Agitator orbital cu incubator Heidolph UNIMAX 1010                          Agitator magnetic Heidolph

         

Bioreactor General Electro 100L and 50 L

  • Drafting the amino acid producing technology
  • Feed and feed additives mixtures and testing them in SPF farms on chickens reared in SPF insulators and on pigeons under trial conditions or other animal species under production conditions

      

Powder and liquid division machinery

  • Assessment of efficacy of products obtained by lactose bioconversion. Invention patent application
  • Demonstration of increase of immune response-mediating pro-inflammatory cytokine synthesis. Testing of certain genetic factors which could interfere with the target gene expression modulation. Invention patent application
  • Assessment of influence of administration of feed additives based on oligosaccharides and chelated microelements on the immune response against certain vaccines particularly in chickens and pigeons. Recording in ROMVAC product list and marketing authorization
  • Web page, promotional materials, articles, international mobility

Step IV – Communication of CO-ROMVAC results

Step V – Communication of CO-ROMVAC results

Step VI – Communication of CO-ROMVAC results

DISSEMINATION

PHASE I

„Whey, a global pollutant, major bioresource for human and veterinary products”, I. Nicolae, C. Chiurciu, V. Chiurciu, P. Sevciuc, D. Pasarin, C. Rovinaru, I.V. Patrascu, P. Stiube, E. Lupu, G. Oltean, G. Motiu, B. Frunzareanu, I. Calin, S, Stanculescu, N. Levandovschi, C. I. Uluitu, A. G. Nica, A. Radu, I. Pais, I. S. Badica, within the Scientific Session “Tradition and innovation in animal health”, Romvac, Voluntari, 29.04.2014

„Comparative study of Kluyveromyces sp. grown on liquid sweet and sour cheese whey”,  Camelia Rovinaru, Diana Pasarin, Ion Nicolae, Bogdan Frunzareanu, Gheorghe Raceanu, Malina Avram, within the International Symposium „Chemistry priorities for Sustainability” PRIOCHEM, 30-31 October 2014, ICECHIM, Bucharest

„Use of a dairy byproduct as substrate for cultivation of yeasts which use lactose as carbon source”, Camelia Rovinaru, Diana Pasarin, Lidia Preda, Luiza Capra, Georgeta Ivan, Malina Avram, within the annual scientific  paper session of the Research-Development Institute for Protection of Plants, 07 November 2014, ASAS, Bucharest

PHASE II

 

   

 Poster –Meat & Milk Conference 3-4 June 2015 Poiana Brasov

„Research for higher whey exploitation”, B.Frunzareanu, I. Nicolae, P. Sevciuc, C.Chiurciu, V. Chiurciu, P. Stiube, D. Pasarin, C. Rovinaru, L. Berca, E. Lupu, S. Stanculescu, A. Nica, V. Tauber, V. Filip, G. Oltean, I. Calin, Expo International Meat&Milk Conference 2015 – Chronicles of an International Success, Brasov, Hotel Alpin, 3-4 June 2015

„Conversion of lactose into microbial biomass, option to reduce organic pollution load of whey”, Diana Pasarin, Camelia Rovinaru, International Conference „Agri-Food sciences, processes and technologies” AGRI-FOOD XXV, 24-25 May 2015,

Sibiu

„Adaptation of a non-conventional yeast strain which metabolize lactose by establishment of culture media composition based on whey”, Camelia Rovinaru, Diana Pasarin, Malina Desliu, International Conference „Agri-Food sciences, processes and technologies” AGRI-FOOD XXV, 24-25 May 2015, Sibiu

„Disruption methods of Saccharomyces sp. yeast cell wall – comparative study”, Diana Pasarin, Camelia Rovinaru, Valentina Filip, Bogdan Frunzareanu, International Symposium „ Chemistry priorities for Sustainability” PRIOCHEM, 29-30 October 2015, ICECHIM, Bucharest

PHASE III

„“Whey pre and pro-biotic nutritional supplements derivatives and their testing on birds in lab and production conditions”, I. Nicolae, C. Chiurciu, V. Chiurciu, P. I. Nicolae, C. Chiurciu, Viorica Chiurciu, P. Sevciuc, P. Stiube, Elena Lupu, B. Frunzareanu, Silvia Stanculescu, Valentina Filip, Lucia Diaconu, Diana Pasarin, CameliaRovinaru, Lavinia Berca, G. Mustatea, G. Oltean, A. Nica, Violeta Tauber, C. Uluitu, N. Levandovschi, Alina Radu, Iulia Calin, S. Gusita,  International Symposium „Chemistry priorities for Sustainability” PRIOCHEM, 27-28 October 2016, ICECHIM, Bucharest

„Extraction of the main polysaccharides from Kluyveromyces marxianus strains cell wall”,  C. Rovinaru, D. Pasarin, International Symposium „Chemistry priorities for Sustainability” PRIOCHEM, 27-28 October 2016, ICECHIM, Bucharest

„Morpho-phenotypic and molecular characterization of Kluyveromyces yeast strains adapted to modified parameters”, Camelia C. Rovinaru, D. Pasarin, L. Berca, International Symposium „Chemistry priorities for Sustainability” PRIOCHEM, 27-28 October 2016, ICECHIM, Bucharest

EVENTS

 During 2-6 November 2016, at INDAGRA, ROMVAC presented in its own stand a poster of OLIGOLAC, as well as samples and leaflets of OLIGOLAC ANIMAL PREMIX and BIOENTEROM at the International Symposium „Chemistry priorities for Sustainability” PRIOCHEM, 27-28 October 2016, ICECHIM, Bucharest

 

CONTACT

Contact person: Dr. Ion NICOLAE, romvac74@gmail.com

Address: Sos. Centurii, nr. 7, Ilfov, 077190, Voluntari

Tel: 021 350 3106, Fax: 021 350 3110