Repository of Research and Investigative Information

Repository of Research and Investigative Information

Baqiyatallah University of Medical Sciences

Application of DNA Shuffling as a Tool for Hydrolase Activity Improvement of Pseudomonas Strain

(2021) Application of DNA Shuffling as a Tool for Hydrolase Activity Improvement of Pseudomonas Strain. Biointerface Research in Applied Chemistry. pp. 7735-7745. ISSN 2069-5837

[img] Text
Application of DNA Shuffling as a Tool for Hydrolase Activity Improvement of Pseudomonas Strain.pdf

Download (418kB)

Official URL: http://apps.webofknowledge.com/InboundService.do?F...

Abstract

Biotechnology is considered one of the most influential technologies in various areas of human life, including health, economics, and the environment. Protein engineering is one of the major biotechnology tools in the field of modification and advancement of biocatalysts capabilities. Among the most effective protein engineering methods, in particular, to improve the industrial strain capabilities, is the shuffling genome method. This study aimed to follow knowledge and biocatalysts engineering techniques based on DNA shuffling methods. In the first step, two procedures were followed (DES method and compatibility according to the concentration gradient of Diazinon) to obtain mutant strains. Acquired mutant strains from both methods were resistant to high concentrations of poison up to 3000 mg/L. The activity of these strains also demonstrated their elevated activity compared to parent samples. The highest activity was related to four strains IR1.G1, IR1.D8, IR1.D4, and IR1.D5, which were 0.234 U/ml, 0.1 U/ml, 0.098 U/ml, and 0.066 U/ml, respectively. The improved strain was obtained via the concentration gradient of the diazinon method (IRL1.G1 strain) in comparison with IRL1.D8 strain (owning highest activity through DES method) possesses excessive activity in 3000 mg/L concentration of Diazinon. The evaluated results of first-generation genome shuffling of strains (the first round of protoplast fusion) also indicated that those shuffled strains with the ability to grow in the vicinity of the toxin (3000 mg/L concentration of Diazinon) showed better activity than obtained mutated strains by both methods (concentration gradient of the toxin and the DES method). In the final stage, the best results were related to IRL1.F2, IRL1.F3, and IRL1.F1 shuffled strains with 0.541 mg/L, 0.523 mg/L, and 0.509 mg/L, respectively. The highest activity belonged to the IRL1.F2 genome shuffled strain (first round of protoplast fusion). This strain could grow in a high concentration of toxin, and also, the activity was increased 30, 3.6, and 2.3 times in comparison with the parent strain (IRL1), IRL.D8 mutant, and IRL1.G1, respectively.

Item Type: Article
Keywords: Diazinon genome shuffling improved strain protein engineering organophosphorus hydrolase biodegradation lactobacillus enzyme gene Chemistry
Page Range: pp. 7735-7745
Journal or Publication Title: Biointerface Research in Applied Chemistry
Journal Index: ISI
Volume: 11
Number: 1
Identification Number: https://doi.org/10.33263/briac111.77357745
ISSN: 2069-5837
Depositing User: مهندس مهدی شریفی
URI: http://eprints.bmsu.ac.ir/id/eprint/9588

Actions (login required)

View Item View Item