FolE gene expression for folic acid productivity from optimized and characterized probiotic Lactobacillus delbrueckii

IF 3.5 Q3 Biochemistry, Genetics and Molecular Biology Journal of Genetic Engineering and Biotechnology Pub Date : 2023-12-18 DOI:10.1186/s43141-023-00603-9
Mohamed Khedr, Fady Sayed Youssef, Noura El-kattan, Mahmoud S. Abozahra, Mohammed N. Selim, Abdullah Yousef, Kamal M. A. Khalil, Alsayed E. Mekky
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Abstract

Lactobacillus delbrueckii was one of the most common milk lactic acid bacterial strains (LAB) which characterized as probiotic with many health influencing properties. Among seven isolates, KH1 isolate was the best producer of folic acid with 100 µg/ml after 48 h of incubation; FolE gene expression after 24 h of incubation was in the highest value in case of KH1 with three folds. Lactose was the best carbon source for this KH1, besides the best next isolates KH80 and KH98. The selected three LAB isolates were identified through 16S rDNA as Lactobacillus delbrueckii. These three isolates have high tolerance against acidic pH 2–3; they give 45, 10, and 22 CFUs at pH 3, besides 9, 6, and 4 CFUs at pH2, respectively. They also have resistance against elevated bile salt range 0.1–0.4%. KH1 recorded 99% scavenging against 97.3% 1000 µg/ml ascorbic acid. Docking study exhibits the binding mode of folic acid which exhibited an energy binding of − 8.65 kcal/mol against DHFR. Folic acid formed four Pi-alkyl, Pi-Pi, and Pi-sigma interactions with Ala9, Ile7, Phe34, and Ile60. Additionally, folic acid interacted with Glu30 and Asn64 by three hydrogen bonds with 1.77, 1.76, and 1.96 Å. LAB isolates have probiotic properties, antioxidant activity, and desired organic natural source for folic acid supplementation that improve hemoglobin that indicated by docking study interaction.
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优化和表征益生菌德尔布鲁贝克氏乳杆菌叶酸生产率的 FolE 基因表达
德尔布鲁贝克乳杆菌(Lactobacillus delbrueckii)是最常见的牛奶乳酸菌(LAB)菌株之一,具有多种影响健康的益生特性。在七个分离株中,KH1 分离株的叶酸产量最高,培养 48 小时后达到 100 µg/ml;培养 24 小时后,KH1 的 FolE 基因表达量最高,达到三倍。乳糖是 KH1 的最佳碳源,其次是 KH80 和 KH98。经 16S rDNA 鉴定,所选的三个 LAB 分离物均为德尔布鲁贝克乳杆菌(Lactobacillus delbrueckii)。这三种分离物对 pH 值为 2-3 的酸性具有很高的耐受性;在 pH 值为 3 时,它们分别能产生 45、10 和 22 个 CFU,而在 pH 值为 2 时,则分别能产生 9、6 和 4 个 CFU。它们还对 0.1-0.4% 的胆盐具有抗性。KH1 对 97.3% 1000 µg/ml 抗坏血酸的清除率为 99%。Docking 研究显示了叶酸的结合模式,它与 DHFR 的结合能量为 - 8.65 kcal/mol。叶酸与 Ala9、Ile7、Phe34 和 Ile60 形成了四种 Pi-alkyl、Pi-Pi 和 Pi-sigma 相互作用。此外,叶酸还与 Glu30 和 Asn64 以 1.77、1.76 和 1.96 Å 的三个氢键相互作用。LAB 分离物具有益生特性和抗氧化活性,是叶酸补充剂的理想有机天然来源,可通过对接研究相互作用改善血红蛋白。
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来源期刊
Journal of Genetic Engineering and Biotechnology
Journal of Genetic Engineering and Biotechnology Biochemistry, Genetics and Molecular Biology-Biotechnology
CiteScore
5.70
自引率
5.70%
发文量
159
审稿时长
16 weeks
期刊介绍: Journal of genetic engineering and biotechnology is devoted to rapid publication of full-length research papers that leads to significant contribution in advancing knowledge in genetic engineering and biotechnology and provide novel perspectives in this research area. JGEB includes all major themes related to genetic engineering and recombinant DNA. The area of interest of JGEB includes but not restricted to: •Plant genetics •Animal genetics •Bacterial enzymes •Agricultural Biotechnology, •Biochemistry, •Biophysics, •Bioinformatics, •Environmental Biotechnology, •Industrial Biotechnology, •Microbial biotechnology, •Medical Biotechnology, •Bioenergy, Biosafety, •Biosecurity, •Bioethics, •GMOS, •Genomic, •Proteomic JGEB accepts
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