DOI: 10.4103/mjbl.mjbl_1185_23 ISSN: 1812-156X
Extraction and Purification of Urease from Helicobacter pylori Isolate from Peptic Ulcer Patients and Study its Optimization via High-Performance Liquid Chromatography
Tamadhur Mohammed Ali Abd Alhasan, Mudher Khudhur Mohammed, Majid Kadhim Abbas Abstract
Background:
Peptic ulcer is a frequent gastrointestinal ailment that causes lesions or ulcers to develop in the stomach, duodenum, or esophageal lining. Infection with
Helicobacter pylori
(
H. pylori
), use of nonsteroidal anti-inflammatory drugs, and smoking are all risk factors for the development of peptic ulcers.
Objective:
The purpose of this study was to screen for and identify
H. pylori
in peptic ulcer patients, as well as to optimize the high-performance liquid chromatography (HPLC) method to purify urease enzyme.
Materials and Methods:
A total of 70 biopsies and stool samples were collected from patients suffering from peptic ulcers, and
H. pylori
infection was determined using both traditional and PCR tests. Ammonium sulfate precipitation was used to partial purification of the urease, and preparative HPLC was used to further purify it.
Results:
The prevalence of
H. pylori
infection was found to be 82.8% based on rapid antigen tests and 17.14% based on bacterial culture. Six isolates were identified by molecular identification using PCR targeting the 16S rRNA gene, confirming their identity as
H. pylori
. The optimum incubation time for maximum urease activity was found to be 72 h. The specific activity of the purified urease was found to be 2.25 U/mg protein, demonstrating a purification fold of 17.17 with a yield of 74.42%.
Conclusion:
High prevalence of
H. pylori
in peptic ulcer patients, and the optimum incubation period for urease production by
H. pylori
was 72 h. The use of HPLC represented an effective technique to purify urease from
H. pylori
.