Primjena rekonstituiranog praha sirutke i mikrobne transglutaminaze u proizvodnji ayrana, tradicionalnog turskog tekućeg jogurta

Research background. In industrial ayran production, milk is diluted to the desired protein content (2% (m/V)) prior to fermentation by yogurt starter cultures by partial replacement of cheese whey in reconstituted form with potable water. This may be an alternative way of protein recovery from cheese by-products as well as reducing the production costs since less milk is used in ayran production. On the other hand, the balance between milk caseins and whey proteins is disturbed when cheese whey is added to milk for ayran production, which likely leads to a time-dependent phase separation during cold storage. Modification of ayran matrix by enzymatic crosslinking of proteins may be a solution to overcome this potential physical instability of ayran. This topic has not been explored to date, and the present study was designed to investigate the possibilities of utilization of reconstituted whey powder (RWP) and microbial transglutaminase (MTG) in ayran production. Experimental approach. Milk was diluted to obtain 2% protein level using RWP and potable water. The aim of utilization of RWP was to meet 5, 10 or 15% of the protein content of the final product. RWP solutions were obtained by calculating the amounts of whey powder required to meet the specified ratios and mixing it with the water required for dilution. We prepared eight different ayran samples divided in three groups, namely group A: prepared by partially diluting milk with RWP to obtain 5, 10 or 15% of total protein amount in the product, group AMTG: prepared by adding microbial transglutaminase (0.5 U per g of protein) to group A samples, and control group without RWP and with or without the addition of MTG. The gross composition, physical (phase separation and viscosity, chemical (volatile and peptide profiles and SDS-PAGE electrophoresis patterns) and sensory properties of the samples were evaluated throughout 15 days of storage with weekly intervals. Results and conclusions. Since the amounts of whey powder used to obtain RWP were different, dry matter levels of the samples differed. Using RWP in ayran production increased the phase separation slightly. Incorporation of MTG affected the physical properties of the ayran samples positively and prevented phase separation at a satisfactory level. SDS-PAGE electrophoretograms revealed that cross-linking between proteins triggered by MTG formed intense bonds at high molecular mass regions. The remaining parameters were not affected by MTG. Results revealed that the samples with 10% RWP with and without addition of MTG were determined as the formulations with the highest commercialization potential. Novelty and scientific contribution. Utilizing RWP in the production of ayran to reduce the protein content of the final product to the desired level is a new approach. Since complete replacement of RWP with potable water to dilute milk to reach 2% (m/V) protein likely leads to lower sensory scores, we have investigated a possibility of partial replacement of RWP with potable water. A time-dependent phase separation is the major challenge of industrial ayran production. This physical problem was largely eliminated by means of MTG-mediated cross-linking of milk proteins. The proposed novel ayran production method offers dairy industry reduction of production costs and contributes to sustainability in milk production since smaller volume of milk is used to reach desired protein content in the final product.Pozadina istraživanja. U industrijskoj proizvodnji ayrana mlijeko se prije fermentacije razrjeđuje do željenog omjera proteina i mlijeka (2 % (m/V)) dodavanjem rekonstituirane sirutke i vode. Osim što se tim postupkom iskorištavaju proteini iz nusproizvoda proizvodnje sira, smanjuju se i troškovi, jer se koristi manji volumen mlijeka za proizvodnju ayrana. S druge strane, dodatak sirutke mlijeku narušava ravnotežu kazeina mlijeka i proteina sirutke, što s vremenom može dovesti do razdvajanja faza tijekom hladnog skladištenja. Modificiranjem matriksa ayrana unakrsnim vezanjem proteina može se smanjiti njegova moguća nestabilnost. Ovaj postupak još nije dovoljno istražen, stoga je u ovom radu ispitana mogućnost primjene rekonstituiranog praha sirutke i mikrobne transglutaminaze u proizvodnji ayrana. Eksperimentalni pristup. Mlijeko je razrjeđeno rekonstituiranim prahom sirutke i vodom kako bi sadržavalo 2 % proteina. Svrha je dodavanja rekonstituiranog praha sirutke bila dobiti konačni proizvod s 5, 10 ili 15 % proteina. Otopine sirutke pripremljene su s izračunatom količinom praha sirutke za postizanje željenog omjera, te miješane s vodom do željenog razrjeđenja. Pripremili smo osam različitih uzoraka ayrana, podijeljenih u tri skupine: skupina A je sadržavala uzorke ayrana s 5, 10 ili 15 % ukupnih proteina pripremljene od djelomično razrjeđenog mlijeka i rekonstituiranog praha sirutke, skupina AMTG pripremljena je dodavanjem 0,5 jedinica mikrobne transglutaminaze po gramu proteina uzorcima iz skupine A, a kontrolna je skupina sadržavala uzorke ayrana bez rekonstituiranog praha sirutke s dodatkom mikrobne transglutaminaze ili bez nje. Sastav, fizikalna (razdvajanje faza i viskoznost), kemijska (sastav hlapljivih spojeva i peptida, te SDS-PAGE profili) i senzorska svojstva uzoraka ispitana su jednom tjedno tijekom 15 dana skladištenja. Rezultati i zaključci. Udjel suhe tvari u uzorcima ayrana se razlikovao, jer su korištene različite količine praha sirutke za dobivanje rekonstituiranog praha. Dodavanjem rekonstituiranog praha sirutke mlijeku tijekom proizvodnje ayrana neznatno se povećalo razdvajanje faza. Ugradnjom mikrobne transglutaminaze u proizvodnju poboljšana su fizikalna svojstva uzoraka te je uspješno zaustavljeno odvajanje faza. Rezultati SDS-PAGE pokazuju da su unakrsnim vezanjem proteina, potaknutim dodatkom mikrobne transglutaminaze, stvorene čvrste veze između molekula velike molekulske mase. Dodatak transglutaminaze nije utjecao na ostale značajke ayrana. Rezultati pokazuju da su uzorci s 10 % rekonstituiranog praha sirutke, bez dodatka ili s dodatkom mikrobne transglutaminaze, okarakterizirani kao proizvodi s najvećim tržišnim potencijalom. Novina i znanstveni doprinos. Primjena rekonstituiranog praha sirutke za smanjenje udjela proteina u krajnjem proizvodu predstavlja novi pristup proizvodnji ayrana. Budući da je potpunom zamjenom rekonstituiranog praha sirutke vodom tijekom razrjeđenja mlijeka radi smanjenja omjera proteina i mlijeka na 2 % rezultiralo lošijim senzorskim ocjenama proizvedenog ayrana, ispitali smo mogućnost djelomične zamjene praha sirutke vodom. Glavni izazov industrijske proizvodnje ayrana je spriječiti odvajanje faza tijekom vremena. Taj se problem može u većoj mjeri riješiti unakrsnim vezanjem proteina pomoću mikrobne transglutaminaze. Predložena nova metoda proizvodnje ayrana omogućuje smanjenje troškova proizvodnje i pridonosi održivosti proizvodnje mlijeka, budući da se za postizanje potrebne količine proteina u krajnjem proizvodu koristi manji volumen mlijeka