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Comparison of in vitro Antifungal Activity Methods Using Extract of Chitinase-producing Aeromonas sp. BHC02

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Abstract

Biological control to prevent fungal plant diseases offers an alternative approach to facilitate sustainable agriculture. Since the chitin in fungal cell walls is a target for biocontrol agents, chitinases are one of the important antifungal molecules. In this study, the aim was to investigate a new chitinase isolated from a fluvial soil bacterium and to show the antifungal activity of the characterized chitinase by comparing the three common methods. The bacterium with the highest chitinase activity was identified as Aeromonas sp. by 16 S rRNA sequence analysis. Following the determination of the optimum enzyme production time, the enzyme was partially purified, and the physicochemical parameters of the enzyme were investigated. In the antifungal studies, direct Aeromonas sp. BHC02 cells or partially purified chitinase were used. As a result, in the first method in which the Aeromonas sp. BHC02 cells were spread on the surface of petri dishes, no zone formation was observed around the test fungi spotted on the surface. However, zone formation was observed in the methods in which the antifungal activity was investigated using the partially purified chitinase enzyme. For example, in the second method, the enzyme was spread on the surface of PDA, and zone formation was observed only around Penicillum species among the test fungi spotted on the surface. In the third method, in which the necessary time was given for the formation of mycelium of the test fungi, it was observed that the growth of Fusarium solani, Alternaria alternata and Botrytis cinerea was inhibited by the partially purified chitinase. This study concludes that the results of the antifungal activities depend on the method used and all fungal chitins cannot be degraded with one strain’s chitinase. Depending on the variety of chitin, some fungi can be more resistant.

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Abbreviations

SDS-PAGE:

Sodium dodecyl sulfate- polyacrylamide gel electrophoresis

NAG:

N-acetylglucosamine

CHD:

Chitinase-detection

NCBI:

National Centre for Biotechnology Information

DNS:

3,5-Dinitrosalicylic acid

BSA:

Bovine serum albumin

V max :

Maximum rate of reaction

K M :

Michaelis Constant

PCR:

Polymerase chain reaction

PDA:

Potato dextrose agar

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Authors and Affiliations

  1. Department of Crop Protection, Tokat Gaziosmanpasa University, Tokat, Turkey

    Bilge Hilal Cadirci

  2. Department of Bioengineering, Hacettepe University, Ankara, Turkey

    Gulesme Yilmaz

  3. Vocational School of Health Services, Medical Laboratory Techniques Fenerbahce University, Istanbul, Turkey

    Gulesme Yilmaz

Authors
  1. Bilge Hilal Cadirci
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Bilge Hilal CADIRCI established the hypothesis of the research, wrote and edit the manuscript. Gulesme Yilmaz did the experiments and prepared almost most of the figures.

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Correspondence to Bilge Hilal Cadirci.

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Cadirci, B.H., Yilmaz, G. Comparison of in vitro Antifungal Activity Methods Using Extract of Chitinase-producing Aeromonas sp. BHC02. Protein J 42, 125–134 (2023). https://doi.org/10.1007/s10930-023-10098-5

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