In vitro production of recombinant ergokonin protein and its application in the polylactic acid film to control Aspergillus niger and Penicillium notatum

Subject Areas : Food Microbiology

Mohammad javad Akbarian Meymand ¹ , Arash Babaei ²

1 - Ph.D. student, Grape Environmental Science Department, Research Institute for Grapes and Raisin (RIGR), Malayer University, Iran.
2 - Assistant Professor, Biology Department, Faculty of Sciences, Malayer University, Malayer, Iran.

Received: 2018-12-17 Accepted : 2019-01-25 Published : 2019-08-23

Keywords: Gene expression, Antimicrobial active packaging, Antifungal combination, Poly-lactic acid film, Ergoconin,

Abstract :

Background & Objectives: Fungal is one of the most important causes of food spoilage. The use of antimicrobial active packaging increases food safety and shelf life. Hence, this study aimed to produce antifungal compounds (Ergoconin) and its application in the polylactic acid film to reduce the fungal contamination of Aspergillus niger and Penicillium notatum.  Materials & Methods: To produce the Ergoconin recombinant protein, the target gene was entered into the pET21 expression vector, and the recombinant vector was transferred to Escherichia coli BL21 (DE3) as host of expression. Gene expression was evaluated by SDS-PAGE, western blot and dot blot. To purify the recombinant protein, a chromatography column was used. Poly-lactic acid bioactive films were prepared by casting method and adding different amounts of Ergoconin (0, 0.25, 0.5, and 0.75%) to 4% wt Poly-lactic acid solution. To investigate the antifungal effect of the poly-lactic acid film, Disc diffusion was used.   Results: Ergokonin gene was replicated by specific primers. The expression of recombinant protein was performed by SDS-PAGE and confirmed by dot and western blot. Ergoconin compound had an antifungal effect on both fungi. The inhibition zone diameter for both fungi increased with increasing Ergoconin concentration. The antimicrobial film had a more antifungal effect on Penicillium notatum.   Conclusion: Ergoconin destroys fungi by preventing the synthesis of glucan in the wall of fungi. Therefore, Poly-lactic acid film containing Ergoconin can be used as antimicrobial active packaging to increase food safety.

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