Molecular Characterization Using gtf Gene Detection and Medium Optimization of Beta-D-Glucan Production from Pediococcus parvulus F1030 Isolated from Local Egyptian Boza
Khalid Abd El Ghany *
National Organization of Drug Control and Research, Giza, Egypt.
Ragaa A. Hamouda
Genetic Engineering and Biotechnology Research Institute, University of Sadat City, Sadat City, Egypt.
Hoda A. Mahrous
Genetic Engineering and Biotechnology Research Institute, University of Sadat City, Sadat City, Egypt.
Sherin M. A. Sharaf
National Organization of Drug Control and Research, Giza, Egypt.
Hanafy A. Hamza
Genetic Engineering and Biotechnology Research Institute, University of Sadat City, Sadat City, Egypt.
*Author to whom correspondence should be addressed.
Abstract
Background: Lactic acid bacteria (LAB) have great benefits which can improve the nutritional value of food, reduces some infections, immune modulator effect, curbs some types of cancer and restricts glucose level of serum.
Aims: This study was carried out to detect glycosyltransferase gene (gtf) encoding glucosyltransferase enzyme responsible for beta-D-glucan production in Pediococcus parvulus F1030 which is isolated by authors from local Egyptian Boza (local alcoholic beverage), and enhancing its production by medium optimization.
Materials and Methods: Molecular identification and DNA sequencing were performed by using specific primers and the medium optimization was enhanced using Plackett-Burman Design.
Results: The isolate detected as Pediococcus parvulus by the molecular identification with using 16S rDNA and submitted in GenBank as Pediococcus parvulus F1030 with accession number ky942131 by the authors. The study revealed the presence of 97% identity with similar to Pediococcus parvulus accession number ky425809. The gtf gene was detected in the isolate by 16S rRNA gene using specific primers. The medium optimization showed enhancement of beta-D-glucan production. The medium number 2 yielded 11 g l-1 from it by using concentration 1.5 g l-1 from sucrose, 0.5 g l-1 from CaCl2, 0.75 g l-1 from tween80, 3 g l-1 from KHPO4 and 1.5 g l-1 from glutamic acid.
Conclusion: Molecular characterization of isolated Pediococcus parvulus showed 97% identity with a similar organism in NCBI site, the detection of gtf gene carried out by using PCR technique with specific primers and beta-d-glucan production was enhanced with medium optimization using Plackett-Burman Design.
Keywords: Pediococcus parvulus, lactic acid bacteria (LAB), gtf gene, beta-d-glucan production, medium optimization.