Abstract
Losses of apple fruit during storage are mainly caused by fungal phytopathogens. Traditionally, postharvest fungal disease is controlled by the application of synthetic fungicides. However, the harmful impact on environment as well as human health largely limits their application. To reduce these problems in agrochemicals usage, new compounds for plant protection, which are eco-friendly, should be developed. The aim of this study is optimization of medium composition in terms of glucose, soybean meal and phosphates content, by applying response surface methodology, for the production of agents with antifungal activity against Aspergillus spp. For biosynthesis was used strain of Streptomyces hygroscopicus isolated from the environment. Experiments were carried out in accordance with Box-Behnken design with three factors on three levels and three repetitions in the central point. Antifungal activity of the obtained cultivation mediums against Aspergillus oryzae and Aspergillus niger was determined, in vitro, using the diffusion - disc method. For determination optimal medium components desirability function was used. Achieved model predicts that the maximum inhibition zone diameter (40.93 mm) against test microorganisms is produced when the initial content of glucose, soybean meal and phosphates are 47.77 g/l, 24.54 g/l and 0.98 g/l, respectively. To minimize the consumption of medium components and costs of effluents processing, additional three sets of optimization were made. The chosen method for optimization of medium components was efficient, relatively simple and time and material saving. Obtained results can be used for the further techno-economic analysis of the process to select optimal medium composition for industrial application.
Highlights
Sterilizacija pripremljenih podloga je izvedena u autoklavu pri temperaturi od 121 oC i pritisku od 2,1 bar u trajanju od 20 min
Biosinteza antifungalnih komponenti u trajanju od 7 dana izvedena je u aerobnim uslovima pri temperaturi od 27 °C i brzini mešanja 150 o/min, koja je obezbeđena postavljanjem erlenmajera na mikrobiološku laboratorijsku tresilicu u termostatu
Po završenoj biosintezi kultivacione tečnosti su centrifugirane pri 10000g u trajanju od 10 min (Eppendorf Centrifuge 5804, Germany)
Summary
Jabuka je jedna od najzastupljenijih voćnih vrsta u svetu i njeni plodovi su dostupni na tržištu tokom cele godine. Vrste roda Aspergillus, pored vrsta rodova Alternaria, Fusarium, Colletotrichum i Penicillium, predstavljaju izazivače bolesti jabuka [3]. S obzirom da tokom patogeneze Aspergillus vrste proizvode različite mikotoksine, najveći problem infekcije je akumulacija toksičnih proizvoda njihovog metabolizma u plodovima jabuke. Proizvodi dobijeni preradom kontaminiranih jabuka su, takođe, izvor mikotoksina jer je veliki broj njih stabilan u ovim procesima [4,5]. Zbog toga je veoma značajno zaštititi jabuke od infekcija izazvanih Aspergillus vrstama. Iako ova tehnika zahteva dosta napora i vremena, najveći nedostatak njene primene je što ne obuhvata interakcije faktora te stoga ne prikazuje ukupan uticaj faktora na proces. Budući da na mikrobiološku proizvodnju antifungalnih komponenti utiče veliki broj faktora, obično je veoma teško utvrditi koji od njih ima najveći uticaj. Za biosintezu upotrebljen je soj Streptomyces hygroscopicus izolovan iz prirodnog okruženja
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