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Applied and Environmental Microbiology, December 2008, p. 7391-7398, Vol. 74, No. 23
0099-2240/08/$08.00+0     doi:10.1128/AEM.01420-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Long-Term Fungal Inhibitory Activity of Water-Soluble Extracts of Phaseolus vulgaris cv. Pinto and Sourdough Lactic Acid Bacteria during Bread Storage{triangledown}

Rossana Coda,1 Carlo G. Rizzello,1 Franco Nigro,1 Maria De Angelis,1 Philip Arnault,2 and Marco Gobbetti1*

Department of Plant Protection and Applied Microbiology, University of Bari, 70126 Bari, Italy,1 Puratos N.V., Industrialaan 25, B-1702 Groot-Bijgaarden, Belgium2

Received 25 June 2008/ Accepted 30 September 2008

The antifungal activity of proteinaceous compounds from different food matrices was investigated. In initial experiments, water-soluble extracts of wheat sourdoughs, cheeses, and vegetables were screened by agar diffusion assays with Penicillium roqueforti DPPMAF1 as the indicator fungus. Water-soluble extracts of sourdough fermented with Lactobacillus brevis AM7 and Phaseolus vulgaris cv. Pinto were selected for further study. The crude water-soluble extracts of L. brevis AM7 sourdough and P. vulgaris cv. Pinto had a MIC of 40 mg of peptide/ml and 30.9 mg of protein/ml, respectively. MICs were markedly lower when chemically synthesized peptides or partially purified protein fractions were used. The water-soluble extract of P. vulgaris cv. Pinto showed inhibition toward a large number of fungal species isolated from bakeries. Phaseolin alpha-type precursor, phaseolin, and erythroagglutinating phytohemagglutinin precursor were identified in the water-soluble extract of P. vulgaris cv. Pinto by nano liquid chromatography-electrospray ionization-tandem mass spectrometry. When the antifungal activity was assayed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, all three proteins were inhibitory. A mixture of eight peptides was identified from the water-soluble extract of sourdough L. brevis AM7, and five of these exhibited inhibitory activity. Bread was made at the pilot plant scale by sourdough fermentation with L. brevis AM7 and addition of the water-soluble extract (27%, vol/wt; 5 mg of protein/ml) of P. vulgaris cv. Pinto. Slices of bread packed in polyethylene bags did not show contamination by fungi until at least 21 days of storage at room temperature, a level of protection comparable to that afforded by 0.3% (wt/wt) calcium propionate.

* Corresponding author. Mailing address: Dipartimento di Protezione delle Piante e Microbiologia Applicata, Via G. Amendola 165/a, 70126 Bari, Italy. Phone: 39 080 5442949. Fax: 39 080 544 29 11. E-mail: gobbetti{at}agr.uniba.it

{triangledown} Published ahead of print on 10 October 2008.

Applied and Environmental Microbiology, December 2008, p. 7391-7398, Vol. 74, No. 23
0099-2240/08/$08.00+0     doi:10.1128/AEM.01420-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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