Deciphering the nature and statistical optimization of antimicrobial metabolites of two endophytic bacilli

Abstract In this study, Allium sativum, garlic, was selected to isolate endophytic bacteria and to evaluate the antimicrobial, antiviral, antioxidant, and cytotoxic activities of their produced metabolites followed by identification of the biosynthetic gene cluster of the antimicrobial metabolites u...

Full description

Saved in:
Bibliographic Details
Main Authors: Raghda S. Isleem, Ahmed M. Eid, Saad El-Din Hassan, Khaled M. Aboshanab, Ghadir S. El-Housseiny
Format: Article
Language:English
Published: SpringerOpen 2025-01-01
Series:AMB Express
Subjects:
Online Access:https://doi.org/10.1186/s13568-024-01811-3
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1832594434587361280
author Raghda S. Isleem
Ahmed M. Eid
Saad El-Din Hassan
Khaled M. Aboshanab
Ghadir S. El-Housseiny
author_facet Raghda S. Isleem
Ahmed M. Eid
Saad El-Din Hassan
Khaled M. Aboshanab
Ghadir S. El-Housseiny
author_sort Raghda S. Isleem
collection DOAJ
description Abstract In this study, Allium sativum, garlic, was selected to isolate endophytic bacteria and to evaluate the antimicrobial, antiviral, antioxidant, and cytotoxic activities of their produced metabolites followed by identification of the biosynthetic gene cluster of the antimicrobial metabolites using Oxford Nanopore Technology (ONT). Two bacterial isolates, C6 and C11, were found to have a broad-spectrum antagonistic effect against four standard microbial strains and were molecularly identified using 16 S ribosomal RNA sequence analysis and deposited in a local culture collection as B. velezensis CCASU-C6, and B. subtilis CCASU-C11, respectively. Optimization for the maximum production of antimicrobial metabolites revealed that a four-day incubation period was optimal, with sucrose and tryptone serving as the best carbon and nitrogen sources for the fermentation media. Response surface methodology model using the central composite design was created resulting in a 1.2-fold and 1.8-fold improvement in antimicrobial metabolite(s) production of C6 and C11 isolates, respectively. The optimal production conditions were found to be a temperature of 33 °C, pH of 7, and an agitation rate of 200 rpm for C6 metabolite, and a temperature of 37 °C, pH of 7, and an agitation rate of 250 rpm for C11 metabolite. Both bacterial isolates displayed antioxidant and antiviral activity and mild cytotoxic action. Genomic sequence and antiSMASH analysis showed that the biosynthetic gene clusters of bacillomycin, mycosubtilin, fengycin, and macrolactin H in B. velezensis CCASU-C6 and bacillibactin and Macrolactin H in B. subtilis CCASU-C11 showed 100% conservation.
format Article
id doaj-art-bad349b710584aef8e15088e0a94ffc9
institution Kabale University
issn 2191-0855
language English
publishDate 2025-01-01
publisher SpringerOpen
record_format Article
series AMB Express
spelling doaj-art-bad349b710584aef8e15088e0a94ffc92025-01-19T12:39:57ZengSpringerOpenAMB Express2191-08552025-01-0115111610.1186/s13568-024-01811-3Deciphering the nature and statistical optimization of antimicrobial metabolites of two endophytic bacilliRaghda S. Isleem0Ahmed M. Eid1Saad El-Din Hassan2Khaled M. Aboshanab3Ghadir S. El-Housseiny4Department of Microbiology and Immunology, Faculty of Pharmacy, Ain Shams UniversityDepartment of Botany and Microbiology, Faculty of Science, Al-Azhar UniversityDepartment of Botany and Microbiology, Faculty of Science, Al-Azhar UniversityDepartment of Microbiology and Immunology, Faculty of Pharmacy, Ain Shams UniversityDepartment of Microbiology and Immunology, Faculty of Pharmacy, Ain Shams UniversityAbstract In this study, Allium sativum, garlic, was selected to isolate endophytic bacteria and to evaluate the antimicrobial, antiviral, antioxidant, and cytotoxic activities of their produced metabolites followed by identification of the biosynthetic gene cluster of the antimicrobial metabolites using Oxford Nanopore Technology (ONT). Two bacterial isolates, C6 and C11, were found to have a broad-spectrum antagonistic effect against four standard microbial strains and were molecularly identified using 16 S ribosomal RNA sequence analysis and deposited in a local culture collection as B. velezensis CCASU-C6, and B. subtilis CCASU-C11, respectively. Optimization for the maximum production of antimicrobial metabolites revealed that a four-day incubation period was optimal, with sucrose and tryptone serving as the best carbon and nitrogen sources for the fermentation media. Response surface methodology model using the central composite design was created resulting in a 1.2-fold and 1.8-fold improvement in antimicrobial metabolite(s) production of C6 and C11 isolates, respectively. The optimal production conditions were found to be a temperature of 33 °C, pH of 7, and an agitation rate of 200 rpm for C6 metabolite, and a temperature of 37 °C, pH of 7, and an agitation rate of 250 rpm for C11 metabolite. Both bacterial isolates displayed antioxidant and antiviral activity and mild cytotoxic action. Genomic sequence and antiSMASH analysis showed that the biosynthetic gene clusters of bacillomycin, mycosubtilin, fengycin, and macrolactin H in B. velezensis CCASU-C6 and bacillibactin and Macrolactin H in B. subtilis CCASU-C11 showed 100% conservation.https://doi.org/10.1186/s13568-024-01811-3EndophytesGarlicBacillomycinMycosubtilinFengycinMacrolactin H
spellingShingle Raghda S. Isleem
Ahmed M. Eid
Saad El-Din Hassan
Khaled M. Aboshanab
Ghadir S. El-Housseiny
Deciphering the nature and statistical optimization of antimicrobial metabolites of two endophytic bacilli
AMB Express
Endophytes
Garlic
Bacillomycin
Mycosubtilin
Fengycin
Macrolactin H
title Deciphering the nature and statistical optimization of antimicrobial metabolites of two endophytic bacilli
title_full Deciphering the nature and statistical optimization of antimicrobial metabolites of two endophytic bacilli
title_fullStr Deciphering the nature and statistical optimization of antimicrobial metabolites of two endophytic bacilli
title_full_unstemmed Deciphering the nature and statistical optimization of antimicrobial metabolites of two endophytic bacilli
title_short Deciphering the nature and statistical optimization of antimicrobial metabolites of two endophytic bacilli
title_sort deciphering the nature and statistical optimization of antimicrobial metabolites of two endophytic bacilli
topic Endophytes
Garlic
Bacillomycin
Mycosubtilin
Fengycin
Macrolactin H
url https://doi.org/10.1186/s13568-024-01811-3
work_keys_str_mv AT raghdasisleem decipheringthenatureandstatisticaloptimizationofantimicrobialmetabolitesoftwoendophyticbacilli
AT ahmedmeid decipheringthenatureandstatisticaloptimizationofantimicrobialmetabolitesoftwoendophyticbacilli
AT saadeldinhassan decipheringthenatureandstatisticaloptimizationofantimicrobialmetabolitesoftwoendophyticbacilli
AT khaledmaboshanab decipheringthenatureandstatisticaloptimizationofantimicrobialmetabolitesoftwoendophyticbacilli
AT ghadirselhousseiny decipheringthenatureandstatisticaloptimizationofantimicrobialmetabolitesoftwoendophyticbacilli