Next Article in Journal
Renieramycin T Induces Lung Cancer Cell Apoptosis by Targeting Mcl-1 Degradation: A New Insight in the Mechanism of Action
Next Article in Special Issue
Characterization of an Alkaline Alginate Lyase with pH-Stable and Thermo-Tolerance Property
Previous Article in Journal
Different Antifungal Activity of Anabaena sp., Ecklonia sp., and Jania sp. against Botrytis cinerea
Previous Article in Special Issue
Microbial Population Changes in Decaying Ascophyllum nodosum Result in Macroalgal-Polysaccharide-Degrading Bacteria with Potential Applicability in Enzyme-Assisted Extraction Technologies
Open AccessArticle

Activity Improvement and Vital Amino Acid Identification on the Marine-Derived Quorum Quenching Enzyme MomL by Protein Engineering

1
College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China
2
Laboratory for Marine Ecology and Environmental Science, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266071, China
3
Institute of Evolution & Marine Biodiversity, Ocean University of China, Qingdao 266003, China
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Mar. Drugs 2019, 17(5), 300; https://doi.org/10.3390/md17050300
Received: 4 April 2019 / Revised: 17 May 2019 / Accepted: 17 May 2019 / Published: 21 May 2019
MomL is a marine-derived quorum-quenching (QQ) lactonase which can degrade various N-acyl homoserine lactones (AHLs). Intentional modification of MomL may lead to a highly efficient QQ enzyme with broad application potential. In this study, we used a rapid and efficient method combining error-prone polymerase chain reaction (epPCR), high-throughput screening and site-directed mutagenesis to identify highly active MomL mutants. In this way, we obtained two candidate mutants, MomLI144V and MomLV149A. These two mutants exhibited enhanced activities and blocked the production of pathogenic factors of Pectobacterium carotovorum subsp. carotovorum (Pcc). Besides, seven amino acids which are vital for MomL enzyme activity were identified. Substitutions of these amino acids (E238G/K205E/L254R) in MomL led to almost complete loss of its QQ activity. We then tested the effect of MomL and its mutants on Pcc-infected Chinese cabbage. The results indicated that MomL and its mutants (MomLL254R, MomLI144V, MomLV149A) significantly decreased the pathogenicity of Pcc. This study provides an efficient method for QQ enzyme modification and gives us new clues for further investigation on the catalytic mechanism of QQ lactonase. View Full-Text
Keywords: quorum quenching enzyme; error prone PCR; high-throughput screening; site-directed mutagenesis; catalytic ability; Pectobacterium carotovorum subsp. carotovorum (Pcc) quorum quenching enzyme; error prone PCR; high-throughput screening; site-directed mutagenesis; catalytic ability; Pectobacterium carotovorum subsp. carotovorum (Pcc)
Show Figures

Figure 1

MDPI and ACS Style

Wang, J.; Lin, J.; Zhang, Y.; Zhang, J.; Feng, T.; Li, H.; Wang, X.; Sun, Q.; Zhang, X.; Wang, Y. Activity Improvement and Vital Amino Acid Identification on the Marine-Derived Quorum Quenching Enzyme MomL by Protein Engineering. Mar. Drugs 2019, 17, 300.

Show more citation formats Show less citations formats
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop