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Microorganisms 2019, 7(3), 74; https://doi.org/10.3390/microorganisms7030074

Response Surface Methodology Optimization of an Acidic Protease Produced by Penicillium bilaiae Isolate TDPEF30, a Newly Recovered Endophytic Fungus from Healthy Roots of Date Palm Trees (Phoenix dactylifera L.)

1
NextBiotech, 98 Rue Ali Belhouane, Agareb 3030, Tunisia
2
Faculty of Science, B.P. 1171, 3000, University of Sfax, Sfax 3029, Tunisia
3
Department of Biology and Genetics, Institute of Biology, Zoology & Radiobiology, University of Veterinary Medicine and Pharmacy, Komenského 73, 04181 Košice, Slovakia
4
Department of Environmental Technology Management, College of Life Sciences, Kuwait University, P.O. Box 5969, Safat 13060, Kuwait
5
Department of Forest Protection, Forest Research Institute, 05-090 Raszyn, Poland
6
Laboratory of Soil Biology, University of Neuchatel, 2000 Neuchatel, Switzerland
*
Author to whom correspondence should be addressed.
Received: 2 February 2019 / Revised: 4 March 2019 / Accepted: 4 March 2019 / Published: 8 March 2019
(This article belongs to the Section Microbial Biotechnology)
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Abstract

To explore proteolytic activity of endophytic fungi inhabiting date palm roots, a Penicillium bilaiae isolate, displaying the highest level of protease production, has been recovered. Response surface methodology (RSM) was applied to optimize culture conditions for protease production by the fungus. Plackett-Burman design allowed for screening of variables effective in protease production. Results indicated that temperature, initial pH and glucose concentration dramatically affect protease yield. These factors were further optimized using a Box-Behnken design and RSM. A combination of initial pH (6.26), temperature (24.5 °C), glucose (13.75 g/L), NaNO3 (1.5 g/L), MgSO4 (0.2 g/L), KH2PO4 (0.5 g/L) and KCl (0.5 g/L) were optimum for maximum production of protease. A 1086-fold enhancement of protease production was gained after optimization. Biochemical properties of fungal protease including the effect of pH and temperature on the activity and the stability of proteolytic enzyme were determined. Moreover, the influence of carbon and nitrogen sources, metal ions, detergents as well as enzyme inhibitors was investigated. Our results highlighted that protease of Penicillium bilaiae isolate TDPEF30 could be considered as a promising candidate for industrial applications. View Full-Text
Keywords: Endophytic fungi; Penicillium bilaiae; protease; response surface methodology; Box-Behnken Endophytic fungi; Penicillium bilaiae; protease; response surface methodology; Box-Behnken
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Ben Mefteh, F.; Frikha, F.; Daoud, A.; Chenari Bouket, A.; Luptakova, L.; Alenezi, F.N.; Al-Anzi, B.S.; Oszako, T.; Gharsallah, N.; Belbahri, L. Response Surface Methodology Optimization of an Acidic Protease Produced by Penicillium bilaiae Isolate TDPEF30, a Newly Recovered Endophytic Fungus from Healthy Roots of Date Palm Trees (Phoenix dactylifera L.). Microorganisms 2019, 7, 74.

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