Director proiect/Project manager: Lector dr. Vamanu Emanuel
Institutie coordonatoare/Coordinator: USAMV Bucuresti, Facultatea de Biotehnologii
Buget proiect/Project budget: 726.600 lei
Rezumatul proiectului/Summary of the project:
La ora actuala, multe specii de ciuperci folosite curent de Medicina Traditionala Chineza si in tarile asiatice, prezinta importante activitati benefice asupra sanatatii umane. De obicei, produsele realizate din aceste ciuperci sunt recunoscute a avea efecte imunomodulatorii, antimicrobiene, antioxidante, şi mai nou diferite efecte antitumorale. Acest proiect intentioneaza sa selecteze din colectia Facultatii de Biotehnologii, tulpinile cele mai productive in scopul obtinerii de biomasa (miceliu) la nivel de bioreactor, in cultivare submersa de tip batch (discontinuu). Aceasta modalitate de obtinere a miceliului este o practica actuala, in biotehnologie, in scopul realizarii cantitatii dorite de biomasa din ciuperci cu proprietati medicinale, fiind una mult mai simpla decat cultivarea in strat solid. Unul din scopurile acestui proiect este elaborarea tehnologiei de obtinere a acestui tip de biomasa in Romania si de realizare a unui produs liofilizat, deoarece produsele de pe piata sunt aduse din import. De asemenea proiectul isi propune, tot ca o noutate, determinarea efectului biomasei liofilizate din ciuperci asupra intestinului uman simulat. Tot la cultivarea in sistem batch, se va urmarii sinteza de exopolizaharide, deoarece reprezinta o componenta importanta a efectului benefic al acestor ciuperci. Se va stabili acea formula de mediu optim pentru cultivare, precum si parametrii ideali de fermentatie la nivel de bioreactor. Dupa separare, si liofilizare, biomasele si produsul se vor analiza din punct de vedere fizico-chimic si microbiologic. Deoarece un tip de produse se bazeaza pe realizarea de extracte din miceliul ciupercilor, proiectul propune realizarea de extracte in pat fluidizat din aceasta biomasa obtinuta la nivel de bioreactor pentru a determina efectul sau antioxidant, de inhibare a diferite tipuri de radicali liberi, precum si evidentierea efectului antimicrobian.
Many species of mushrooms currently used in traditional chinese medicine and in asian countries have significant beneficial influences upon human health. Usually, the products obtained from these mushrooms are known to have immuno-modulatory, antimicrobial, antioxidant and more recently determined various anti-tumoral effects. The purpose of this project is to select form the collection of the faculty of biotechnologies the most productive strains in order to obtain biomass (mycelium) in bioreactors, by batch submerse cultivation (discontinuous). This method for obtaining mycelium is a current practice in biotechnology, intended to produce the required quantity of fungal biomass with medicinal properties, much simpler than solid substrate cultivation. One of the objectives of this project is to elaborate the technology for obtaining this type of biomass in romania and to create a lyophilized product, because those existing on the market are imported. Also as a novelty, the project aims at determining the effect of fungal lyophilized biomass upon simulated human intestine. Besides, during batch cultivation, the exopolysaccharide synthesis will be observed, as it represents an important aspect of the beneficial effect of these fungi. The optimal culture medium formula and the ideal fermentation parameters in bioreactors will be determined. After separation and lyophilization, the biomasses and the product will be subjected to physicochemical and microbiological analysis. Considering that a type of products is based on fungal mycelium extracts, the project aims at creating fluidized bed extracts of such biomass obtained in a bioreactor, in order to determine its antioxidant effect, of inhibition of various free-radical types, and to highlight the antimicrobial effect.
Obiectivele generale si specifice ale proiectului urmaresc/The general and the specific objectives of the project are:
Obiectivele generale si specifice ale proiectului urmaresc: elaborarea si optimizarea metodelor si tehnologiilor avansate de identificare a tulpinilor de ciuperci din specia Pleurotus ostreatus; multiplicarea in conditii de laborator si bioreactor in vederea obtinerii unei productii ridicate de exopolizaharide cu rol semnificativ in cresterea proprietatilor reologice; selectia de tulpini cu valoare medicinala ridicata, testate cu ajutorul unui sistem de simulare a tractului digestiv uman; activitati de conditionare – liofilizare si realizare de extracte prin mijloace de ultima generatie – extractie in pat fluidizat; actiuni-suport: activitati specifice de diseminare.Obiectivul general este dezvoltarea unor biotehnologii avansate de obtinere la nivel de bioreactor si liofilizare a biomasei de Pleurotus ostreatus, sinteza a exopolizaharidelor, biotehnologii extractive adaptate ultimelor noutati in domeniu.
The general and the specific objectives of the project are: the development and optimization of advanced methods and technologies for identification of fungal strains of Pleurotus ostreatus species; themultiplication in laboratory and bioreactor conditions in order to obtain a high production of exopolysaccharide significant in increasing rheological properties; selection of strains with high medicinal value, tested by means of a simulation system of human digestive tract; conditioning activities – lyophilization and creation of extracts by most recent means - fluidized bed extraction; support activities: specific activities of dissemination and training activities with students and post-graduates attending master’s studies of the Faculty of Biology and Biotechnology. The general objective is to develop advanced biotechnologies for obtaining Pleurotus ostreatus biomass in bioreactors by lyophilization, synthesis of exopolysaccharide, extractive biotechnologies adapted to the latest news in the domain.
Obiectivele specifice vor fi solutionate in acord cu planul de realizare al proiectului:
The specific objectives will be settled according to the plan of project accomplishment:
- Studiu teoretic privind utilizarea ciupercii Pleurotus ostreatus, respectiv a metodelor de analiza fizico – chimica si a posibilei structurii a acestora. Identificarea si caracterizarea tulpinilor aflate in colectia facultatii.
- Theoretical study on the use of Pleurotus ostreatus, the physicochemical analysis methods respectively, and the possibility of their organization; identification and characterization of strains in the collection of the faculty.
- Incadrarea taxonomica a tulpinilor izolate prin tehnici conventionale si de biologie moleculara. Stabilirea compozitiei mediilor de cultura pentru cultivarea ciupercii la nivel de laborator si bioreactor. Stabilirea fluxului tehnologic de obtinere a biomaselor de Pleurotus ostreatus. Evidentierea sintezei de exopolizaharide la nivel de bioreactor si izolarea acestora. Liofilizarea biomasei in diferite medii crioprotectoare. Analiza fizico-chimica si microbiologica a liofilizatelor obtinute.
- Taxonomical classification of isolated strains by conventional techniques and by those of molecular biology; determination of culture media composition for fungal cultivation in laboratories and bioreactors; establishment of technological flow for obtaining Pleurotus ostreatus biomass; highlighting exopolysaccharide synthesis in bioreactors and their isolation; lyophilization of biomass in various cryoprotectant media; physicochemical and microbiological analysis of the lyophilized obtained.
- Realizarea de extracte cu extractorul in pat fluidizat si diferiti solventi. Analiza activitatii antioxidante si de legate a diferitelor tipuri de radicali liberi. Evidentierea efectului antimicrobian al extractului din Pleurotus ostreatus. Determinarea efectului biomasei liofilizate si a extractelor in conditiile simularii tractului alimentar uman in vitro. Realizarea unui produs bazat pe biomasa liofilizata de Pleurotus ostreatus si analiza sa fizico-chimica si microbiologica. Brevetare.
- Obtaining extracts with fluidized bed extractor and various solvents; analysis of antioxidant activities and those of binding various types of free radicals; highlighting antimicrobial effect of Pleurotus ostreatus extract; determination of the effect of lyophilized biomass and extracts while simulating human alimentary tract in vitro. Obtaining a product based on lyophilized biomass of Pleurotus ostreatus and its physicochemical and microbiological analysis. Patent.