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Production of Phytase Enzyme by a Bioengineered Probiotic for Degrading of Phytate Phosphorus in the Digestive Tract of Poultry

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Abstract

Probiotics are beneficial microorganisms and have long been used in food production as well as health promotion products. Bioengineered probiotics are used to express and transfer native or recombinant molecules to the mucosal surface of the digestive tract to improve feed efficiency and promote health. Lactococcus lactis is a potential probiotic candidate to produce useful biological proteins. The aim of this investigation was to develop a recombinant Lactococcus lactis with the potential of producing phytase. To enhance the efficiency of expression and secretion of recombinant phytase, usp45 signal peptide was added to the expression vector containing phytase gene (appA2) derived from Escherichia coli. Sequencing of recombinant plasmid containing appA2 showed the correct construction of plasmid. Total length of the phytase insert was 1.25 kbp. A Blast search of the cloned fragment showed 99% similarity to the reported E. coli phytase sequence in the GenBank (accession number: AM946981.2). A plasmid containing usp45 and appA2 electrotransferred into Lactococcus lactis. Zymogram with polyacrylamide gel revealed that the protein extract from the supernatant and the cell pellet of recombinant bacteria had phytase activity. Enzyme activity of 4 U/ml was obtained in cell extracts, and supernatant maximal phytase activity was 19 U/ml. The recombinant L. lactis was supplemented in broiler chicken feed and showed the increase of apparent digestibility on phytate phosphorus in the digestive tract and it was same as performance of E. coli commercial phytase.

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Acknowledgments

The authors gratefully acknowledge the financial support from the Excellence Centre for Animal Sciences, and Faculty of Agriculture, Ferdowsi University of Mashhad and Dr. Banayan Aval for his English editing service.

Funding

This study was funded by Ferdowsi university of Mashhad (FUM) 3/30173; 2/16/2014.

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Authors and Affiliations

  1. Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran

    Bahareh Pakbaten, Reza Majidzadeh Heravi, Hassan Kermanshahi, Mohammad-Hadi Sekhavati, Ali Javadmanesh & Masoud Mohammadi Ziarat

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  1. Bahareh Pakbaten
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  2. Reza Majidzadeh Heravi
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  3. Hassan Kermanshahi
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Contributions

BP carried out experimental process. RM designed and drafted the manuscript. HK and MS analyzed data and information of study. AJ conducted the study in laboratory. MM carried out animal model part of study. All author contributed in interpretation of results and read and approved the final manuscript.

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Correspondence to Reza Majidzadeh Heravi.

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The authors declare that they have no conflict of interest.

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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

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Pakbaten, B., Majidzadeh Heravi, R., Kermanshahi, H. et al. Production of Phytase Enzyme by a Bioengineered Probiotic for Degrading of Phytate Phosphorus in the Digestive Tract of Poultry. Probiotics & Antimicro. Prot. 11, 580–587 (2019). https://doi.org/10.1007/s12602-018-9423-x

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