## 5 Conclusions  
In conclusion, the MLH at MICs can inhibit isoleucyl-tRNA synthetase in MRSA as well as muiprocin only as observed by CLSM. The MICs of muiprocin only and MLH were able to trigger a stringent response in MRSA which led to the up- and down-regulation of genes during the exponential phase of MRSA growth. Transcriptome profiles of MRSA showed that global transcriptional alterations after treatment with muiprocin were similar to those treated with MLH at MICs, including genes involved in the regulation of transcription, translation, and replication pathways, though the fold changes in these genes decreased more rapidly with MLH than with muiprocin after only 1 h of treatment. The inhibition of muiprocin and MLH prevented incorporation of isoleucine into the protein chains, resulting in inhibition of protein synthesis, whereas induction of GTP resulted in growth inhibition. Finally, MLH can be successfully developed for use as an antibacterial skin formulation and can replace muiprocin ointment. A future study will focus on an animal model for studies of safety and efficacy.  
The following supporting information can be downloaded at: [[https://www.mdpi.com/article/10.3390/applmicrobioI2010019/s1](https://www.mdpi.com/article/10.3390/applmicrobioI2010019/s1)]([https://www.mdpi.com/article/10.3390/applmicrobioI2010019/s1](https://www.mdpi.com/article/10.3390/applmicrobioI2010019/s1)), Table S1: Preparation of MLH, Table S2: Decreases in the expression of genes involved in important functions following exposure to MICs of muiprocin and MLH compared with the control at 5, 6, and 12 h, Table S3: Increases in the expression of genes involved in important functions following exposure to MICs of muiprocin and MLH compared with the control at 5, 6, and 12 h, Table S4: Increases in the expression of genes involved in important functions following exposure to MICs of muiprocin and MLH at 5 h and decreases in gene expression at 6 and 12 h compared with the control, Table S5: Decreases in the expression of genes involved in important functions following exposure to MICs of muiprocin and MLH at 5 and 6 h and increases in gene expression at 12 h compared with the control.  
Conceptualization, S.K. and T.S.; investigation, S.K., M.W. and P.D.; resources, M.W. and T.S.; data analysis, K.S. and S.K.; writing--original draft preparation, S.K.; writing--review and editing, T.S.; visualization, S.K.; supervision, T.S.; project administration, T.S.; funding acquisition, S.K. and T.S. All authors have read and agreed to the published version of the manuscript.  
This work was supported by the Royal Golden Jubilee Ph.D. program of the Thailand Research Fund; PHD/0088/2559 for [PERSON] and the National Research Council of Thailand (PHA610372S).  
Not applicable.  
The sequence read archive data are available from the National Center for Biotechnology Information under accession number PRJNA634329. The data can be accessed using this link: [[http://www.ncbi.nlm.nih.gov/bioproject/634329](http://www.ncbi.nlm.nih.gov/bioproject/634329)]([http://www.ncbi.nlm.nih.gov/bioproject/634329](http://www.ncbi.nlm.nih.gov/bioproject/634329)) (accessed on 21 January 2022).  
This research was supported by the Royal Golden Jubilee Ph.D. program of the Thailand Research Fund; PHD/0088/2559 for [PERSON]. [PERSON] and the National Research Council of Thailand (PHA610372S). We are grateful to the Drug Delivery System Excellence Center (DDESC), Faculty of Pharmaceutical Sciences, Prince of Songkla University for the use of research facilities. We would like to thank [PERSON] for her English editing.  
The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript.