LAPSE:2024.1709v1
Published Article
LAPSE:2024.1709v1
Expression, Characterization, and Immobilization of a Novel D-Lactate Dehydrogenase from Salinispirillum sp. LH 10-3-1
August 23, 2024
Abstract
sp. LH 10-3-1 was newly isolated from the alkali lake water samples collected in Inner Mongolia. In this study, a gene coding for D-lactate dehydrogenase from the strain LH 10-3-1 (SaLDH) was cloned and characterized. The recombinant enzyme was a tetramer with a native molecular mass of 146.2 kDa. The optimal conditions for SaLDH to reduce pyruvate and oxidize D-lactic acid were pH 8.0 and pH 5.0, at 25 °C. Cu2+ and Ca2+ slightly promoted the oxidation and reduction activities of SaLDH, respectively. To improve the stability of SaLDH, the enzyme was immobilized on Cu3(PO4)2-based inorganic hybrid nanoflowers. The results showed that the reduction activity of the hybrid nanoflowers disappeared, and the optimum temperature, specific activity, thermostability, and storage stability of the immobilized SaLDH were significantly improved. In addition, the biotransformation of D-lactic acid to pyruvate catalyzed by SaLDH and the hybrid nanoflowers was investigated. The maximum conversion of D-lactic acid catalyzed by the immobilized SaLDH was 25.7% higher than by free enzymes, and the immobilized SaLDH could maintain 84% of its initial activity after six cycles.
sp. LH 10-3-1 was newly isolated from the alkali lake water samples collected in Inner Mongolia. In this study, a gene coding for D-lactate dehydrogenase from the strain LH 10-3-1 (SaLDH) was cloned and characterized. The recombinant enzyme was a tetramer with a native molecular mass of 146.2 kDa. The optimal conditions for SaLDH to reduce pyruvate and oxidize D-lactic acid were pH 8.0 and pH 5.0, at 25 °C. Cu2+ and Ca2+ slightly promoted the oxidation and reduction activities of SaLDH, respectively. To improve the stability of SaLDH, the enzyme was immobilized on Cu3(PO4)2-based inorganic hybrid nanoflowers. The results showed that the reduction activity of the hybrid nanoflowers disappeared, and the optimum temperature, specific activity, thermostability, and storage stability of the immobilized SaLDH were significantly improved. In addition, the biotransformation of D-lactic acid to pyruvate catalyzed by SaLDH and the hybrid nanoflowers was investigated. The maximum conversion of D-lactic acid catalyzed by the immobilized SaLDH was 25.7% higher than by free enzymes, and the immobilized SaLDH could maintain 84% of its initial activity after six cycles.
Record ID
Keywords
characterization, D-lactate dehydrogenase, expression, immobilization, Salinispirillum sp.
Subject
Suggested Citation
Liu J, Jiang X, Zheng Y, Li K, Zhang R, Xu J, Wang Z, Zhang Y, Yin H, Li J. Expression, Characterization, and Immobilization of a Novel D-Lactate Dehydrogenase from Salinispirillum sp. LH 10-3-1. (2024). LAPSE:2024.1709v1
Author Affiliations
Liu J: Department of Biological and Bioenergy Chemical Engineering, College of Chemistry and Chemical Engineering, China University of Petroleum (East China), Qingdao 266580, China
Jiang X: Department of Biological and Bioenergy Chemical Engineering, College of Chemistry and Chemical Engineering, China University of Petroleum (East China), Qingdao 266580, China
Zheng Y: Department of Biological and Bioenergy Chemical Engineering, College of Chemistry and Chemical Engineering, China University of Petroleum (East China), Qingdao 266580, China
Li K: Department of Biological and Bioenergy Chemical Engineering, College of Chemistry and Chemical Engineering, China University of Petroleum (East China), Qingdao 266580, China
Zhang R: Department of Biological and Bioenergy Chemical Engineering, College of Chemistry and Chemical Engineering, China University of Petroleum (East China), Qingdao 266580, China
Xu J: Department of Biological and Bioenergy Chemical Engineering, College of Chemistry and Chemical Engineering, China University of Petroleum (East China), Qingdao 266580, China
Wang Z: Department of Biological and Bioenergy Chemical Engineering, College of Chemistry and Chemical Engineering, China University of Petroleum (East China), Qingdao 266580, China
Zhang Y: Department of Biological and Bioenergy Chemical Engineering, College of Chemistry and Chemical Engineering, China University of Petroleum (East China), Qingdao 266580, China
Yin H: Department of Biological and Bioenergy Chemical Engineering, College of Chemistry and Chemical Engineering, China University of Petroleum (East China), Qingdao 266580, China
Li J: Department of Biological and Bioenergy Chemical Engineering, College of Chemistry and Chemical Engineering, China University of Petroleum (East China), Qingdao 266580, China
Jiang X: Department of Biological and Bioenergy Chemical Engineering, College of Chemistry and Chemical Engineering, China University of Petroleum (East China), Qingdao 266580, China
Zheng Y: Department of Biological and Bioenergy Chemical Engineering, College of Chemistry and Chemical Engineering, China University of Petroleum (East China), Qingdao 266580, China
Li K: Department of Biological and Bioenergy Chemical Engineering, College of Chemistry and Chemical Engineering, China University of Petroleum (East China), Qingdao 266580, China
Zhang R: Department of Biological and Bioenergy Chemical Engineering, College of Chemistry and Chemical Engineering, China University of Petroleum (East China), Qingdao 266580, China
Xu J: Department of Biological and Bioenergy Chemical Engineering, College of Chemistry and Chemical Engineering, China University of Petroleum (East China), Qingdao 266580, China
Wang Z: Department of Biological and Bioenergy Chemical Engineering, College of Chemistry and Chemical Engineering, China University of Petroleum (East China), Qingdao 266580, China
Zhang Y: Department of Biological and Bioenergy Chemical Engineering, College of Chemistry and Chemical Engineering, China University of Petroleum (East China), Qingdao 266580, China
Yin H: Department of Biological and Bioenergy Chemical Engineering, College of Chemistry and Chemical Engineering, China University of Petroleum (East China), Qingdao 266580, China
Li J: Department of Biological and Bioenergy Chemical Engineering, College of Chemistry and Chemical Engineering, China University of Petroleum (East China), Qingdao 266580, China
Journal Name
Processes
Volume
12
Issue
7
First Page
1349
Year
2024
Publication Date
2024-06-28
ISSN
2227-9717
Version Comments
Original Submission
Other Meta
PII: pr12071349, Publication Type: Journal Article
Record Map
Published Article
LAPSE:2024.1709v1
This Record
External Link
https://doi.org/10.3390/pr12071349
Publisher Version
Download
Meta
Record Statistics
Record Views
475
Version History
[v1] (Original Submission)
Aug 23, 2024
Verified by curator on
Aug 23, 2024
This Version Number
v1
Citations
Most Recent
This Version
URL Here
http://psecommunity.org/LAPSE:2024.1709v1
Record Owner
PSE Press
Links to Related Works