Phytase superior Lactobacillusisolate (L-Phy5) and its mutants: molecular identification | ||
| Egyptian Pharmaceutical Journal | ||
| Volume 24, Issue 4, October 2025, Pages 104-120 PDF (974.57 K) | ||
| Document Type: Original Article | ||
| DOI: 10.21608/epj.2025.405118.1152 | ||
| Author | ||
| Alawiah M Alhebshi* | ||
| Biological Science Department- faculty of science-King Abdul-Aziz University-Jeddah- Saudi Arabia | ||
| Abstract | ||
| Background Phytase produced by lactic acid bacteria (LAB), which are classified as generally recognized as safe (GRAS), used in food processing. Phytase produced by Lactobacillus plantarum and acidophilus, degrade phytic acid —a common antinutrient in plant-based foods, offers several key benefits, in food to improve nutrient bioavailability. Phytic acid binds essential minerals like iron, zinc, and calcium, reducing their availability for absorption. Phytase helps release these bound minerals, for absorption in the gut. Objective The present study aimed to isolate a probiotic strain that excels in phytase synthesis, identify it at the molecular level, and assess its efficacy as a probiotic. Mutagenesis and protoplast fusion used in a genetic improvement program to create mutants and fusants with superior enzyme output. Materials and methods Different isolates of Lactobacillus were isolated, examined for phytase synthesis and the good candidate for phytase synthesis was identified using 16SrRNA. To enhance its phytase production, a genetic improvement using physical mutagenesis via ultraviolet (UV) radiation and chemical mutagenesis with ethyl methanesulfonate (EMS) used. The protoplast fusion between two mutants (PUV12-10 and PMS60-5) was performed to enhance phytase production. The molecular diversity induced by EMS and UV mutagenesis, as well as protoplast fusion in Lactobacillus reuteri Pro8 (L-Phy5), using RAPD analysis with 4 random primers. Results and conclusion Seven Lactobacillus isolates were isolated with one (L-Phy5) identified as Lactobacillus reuteri Strain Pro8 (PV448931.1), a good candidate for phytase synthesis. strong probiotic potential, implying its feasibility for incorporation into functional food items. To enhance its phytase production, a genetic improvement program (UV) radiation and (EMS) was used. Both UV-treated mutant (PUV12-10) and EMS-induced mutants (PMS60-5) exhibited (258.76% and (287.53%) phytase increase compared to the parent strain. The protoplast fusion between (PUV12-10 and PMS60-5) successfully yielded fusant (PC1/9)as a promising candidate for further development in phytase-based biotechnological applications. The molecular diversity induced by EMS and UV mutagenesis, as well as protoplast fusion in Lactobacillus reuteri Pro8 (L-Phy5), using RAPD analysis. Distinct banding patterns were observed among EMS-induced mutants, recombinant fusants, and the parental strain. RAPD-based phylogenetic analysis revealed distinct genetic relationships among Lactobacillus reuteri L-Phy5, its mutants, and fusants | ||
| Keywords | ||
| Keywords: Lactobacillus reuteri; phytase; mutants; fusants; phylogenetic; random amplification of polymorphic DNA Polymerase Chain Reaction (RAPD-PCR) | ||
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