Bacillus Metabolites: Compounds, Identification and Anti-Candida albicans Mechanisms
Round 1
Reviewer 1 Report
In the reviewed manuscript, authors discuss about the application of Bacillus metabolites in the control of C. albicans. This review started with compounds produced by Bacillus spp., described the isolation and identification techniques of Bacillus metabolites and summarized their mechanisms against C. albicans. In addition to conventional separation and identification methods, high-precision separation technology and omics technology played an important role in the separation of similar components of Bacillus metabolites. And from the inhibition of pathogenic fungi themselves and inhibition of the virulence factors of C. albicans two aspects, the antibacterial mechanism of Bacillus metabolites on C. albicans were described. This review is expected to bring positive significance for the control of pathogenic fungi such as C. albicans and the application of Bacillus metabolites in the future. In general, the review represents a very big piece of information. The topic is interesting and well within the aims of the Journal, but it needs minor revisions before it ca be published and the authors need to address the following issues before it can be accepted for publication.
1. I have read the entire manuscript and my initial comment is that manuscript is well-written. The structure of the abstract should be improved, please give more precise objectives here (such as in the Abstract).
2. Keywords must be arranged in alphabetic order and are written with capital initials, others with lowercase initials, please make them uniform. Punctuation also needs to be fixed.
- The author must add a figure to give an overview of Bacillus metabolites or compounds, Identification, and their working principle or how they act as anti-fungal. I mean author gives a regulatory or working mechanism for these metabolites.
Author Response
Point 1: I have read the entire manuscript and my initial comment is that manuscript is well-written. The structure of the abstract should be improved, please give more precise objectives here (such as in the Abstract).
Response 1: Thank you for your helpful advice. We have re-written some sentences of the abstract. The original and revised versions of the Abstract are attached below.
Original version of the Abstract:
Candida albicans is a serious threat to human health, especially for immunocompromised groups. The drugs used to treat fungal infections include azoles, polyenes and echinocandins. However, the limited types of antifungal drugs and the widespread emergence of drug resistance have increased the incidence of fungal infections. Therefore, it is greatly significant to develop new antifungal drugs. Bacillus and its metabolites have antifungal activity against pathogenic fungi. This review aims to introduce the application of Bacillus metabolites in the control of C. albicans in recent years. The review started with compounds produced by Bacillus spp., described the isolation and identification techniques of Bacillus metabolites in recent years, and summarized their mechanisms against C. albicans. In addition to conventional separation and identification methods, high-precision separation technology and omics technology played an important role in the separation of similar components of Bacillus metabolites. And from the inhibition of pathogenic fungi themselves and inhibition of the virulence factors of C. albicans two aspects, the antifungal mechanism of Bacillus metabolites on C. albicans were described. This review is expected to become the reference for the control of pathogenic fungi such as C. albicans and the application of Bacillus metabolites in the future.
Revised versions of the Abstract:
Candida albicans seriously threatens human health, especially for immunosuppressed groups. The antifungal agents mainly include azoles, polyenes and echinocandins. However, the few types of existing antifungal drugs and their resistance make it necessary to develop new antifungal drugs. Bacillus and its metabolites have antifungal activity against pathogenic fungi. This review introduced the application of Bacillus metabolites in the control of C. albicans in recent years. Firstly, several compounds produced by Bacillus spp. were listed. Then the isolation and identification techniques of Bacillus metabolites in recent years were described, including high-precision separation technology and omics technology for the separation of similar components of Bacillus metabolites. And the mechanisms of Bacillus metabolites against C. albicans were summarized from the inhibition of pathogenic fungi themselves and inhibition of the fungal virulence factors. The purpose of this review is to systematically summarize the recent studies on the inhibition of pathogenic fungi by Bacillus metabolites. The review is expected to become the reference for the control of pathogenic fungi such as C. albicans and the application of Bacillus metabolites in the future.
Point 2: Keywords must be arranged in alphabetic order and are written with capital initials, others with lowercase initials, please make them uniform. Punctuation also needs to be fixed.
Response 2: Keywords have been arranged in alphabetic order and written with capital initials, others with lowercase initials. We made them uniform and fixed the punctuation.
Point 3: The author must add a figure to give an overview of Bacillus metabolites or compounds, Identification, and their working principle or how they act as anti-fungal. I mean author gives a regulatory or working mechanism for these metabolites.
Response 3: We have added a figure (Figure 4, Page 10 line 420-423 in revised version) which described the classification, purification and identification, and antifungal mechanisms of Bacillus metabolites. The introduction text of this figure was also added (Page 10 line 434-435 in revised version).
Author Response File: 
Author Response.docx
Reviewer 2 Report
It is very interesting work. Written carefully.
This work can be published after minor corrections.
Minors:
Page 1, line 31; “Candida albicans” should be “C. albicans”?
Page 2, line 67-68; The structures of “surfactin, iturin and fengycin” should be given.
Page 2, line 82; “B. aryabhattai” should be italic.
Page 3, line 100-101; The structures of “bacillaene, difficidin, and macrolactin” should be given.
Page 3, line 108-109; The structures of “lanthionine, subtilin, and nisin A” should be given.
Page 5, line 213; “4.1. Purification and identification technology of Bacillus metabolites” is the same as “3.1. Purification and identification technology of Bacillus metabolites”?
Author Response
Point 1: Minors:
Page 1, line 31; “Candida albicans” should be “C. albicans”?
Page 2, line 67-68; The structures of “surfactin, iturin and fengycin” should be given.
Page 2, line 82; “B. aryabhattai” should be italic.
Page 3, line 100-101; The structures of “bacillaene, difficidin, and macrolactin” should be given.
Page 3, line 108-109; The structures of “lanthionine, subtilin, and nisin A” should be given.
Page 5, line 213; “4.1. Purification and identification technology of Bacillus metabolites” is the same as “3.1. Purification and identification technology of Bacillus metabolites”?
Response 1: Thanks for your careful review. Some statements have been modified to improve originality, also added figures, so the position of some words has changed. The modifications listed below are in the pages and lines of the revised version.
Page 1, line 38; “Candida albicans” was corrected to “C. albicans”.
Page 3, line 87-88; The structures of “surfactin, iturin and fengycin” were shown as Figure 1 (Reference 12. Caulier, S.; Nannan C.; Gillis A.; Licciardi F.; Bragard C.; Mahillon J. Overview of the Antimicrobial Compounds Produced by Members of the Bacillus subtilis Group. Front. Microbiol., 2019, 10: 302.) and the corresponding explanation was also added in the text (Page 2, line 75).
Page 3, line 94; The sentence was re-written and “Bacillus aryabhattai” was italic.
Page 4, line 133-135; The structures of “bacillaene, difficidin, and macrolactin” were shown as Figure 2 (Reference 12. Caulier, S.; Nannan C.; Gillis A.; Licciardi F.; Bragard C.; Mahillon J. Overview of the Antimicrobial Compounds Produced by Members of the Bacillus subtilis Group. Front. Microbiol., 2019, 10: 302.) and the corresponding explanation was also added in the text (Page 3, line 121).
Page 4, line 152-Page 5, line 155; The structures of “lanthionine, subtilin, and nisin A” were shown as Figure 3 (Reference 12. Caulier, S.; Nannan C.; Gillis A.; Licciardi F.; Bragard C.; Mahillon J. Overview of the Antimicrobial Compounds Produced by Members of the Bacillus subtilis Group. Front. Microbiol., 2019, 10: 302.) and the corresponding explanation was also added in the text (Page 4, line 138).
Page 4, line 261; “4.1. Purification and identification technology of Bacillus metabolites” has been corrected to “4.1. Inhibition of pathogenic fungi”.
Author Response File: Author Response.docx
Reviewer 3 Report
The plagiarism is 36%. Please reduce. refer PDF files.
English language correction is required, a few corrections are mentioned below:
Line 7: immunosuppressed instead of immunocompromised
line 34, in patients remove the
line 36: targets limit
line 51: remove "it was found that"
line 58: has "a" fast reproduction
LINE 60 : adaptability and a resistance to stress,....
line 135: have 'a' strong ability to inhibit
line 184: which can 'perform' efficient separation
line201: The researchers developed an integrated
line 204: and analysis of NMR data
line 220 : 'The' cell wall is the first target
line 226 : the improve fluconazole efficacy [42]
line 275: The hyphal state of C. albicans has a stronger ability for tissue invasion and infiltration.
Comments for author File: 
Comments.pdf
Author Response
Point 1: The plagiarism is 36%. Please reduce. refer PDF files.
Response 1: Thanks for your helpful review. Refer to PDF files, we have modified some sentences to improve originality. All the modified content was marked in revision mode in the text. The modifications made are as follows. The modifications listed below are in the lines of the revised version.
Line 7-12: “Candida albicans is a serious threat to human health, especially for immunocompromised groups. The drugs used to treat fungal infections include azoles, polyenes and echinocandins. However, the limited types of antifungal drugs and the widespread emergence of drug resistance have increased the incidence of fungal infections. Therefore, it is greatly significant to develop new antifungal drugs.” was revised as “Candida albicans seriously threatens human health, especially for immunosuppressed groups. The antifungal agents mainly include azoles, polyenes and echinocandins. However, the few types of existing antifungal drugs and their resistance make it necessary to develop new antifungal drugs.”
Line 13: “aims to” was removed. “introduce” was changed to “introduced”.
Line 14-26:
“The review started with compounds produced by Bacillus spp., described the isolation and identification techniques of Bacillus metabolites in recent years, and summarized their mechanisms against C. albicans. In addition to conventional separation and identification methods, high-precision separation technology and omics technology played an important role in the separation of similar components of Bacillus metabolites. And from the inhibition of pathogenic fungi themselves and inhibition of the virulence factors of C. albicans two aspects, the antifungal mechanism of Bacillus metabolites on C. albicans were described. This review is expected to become the reference for the control of pathogenic fungi such as C. albicans and the application of Bacillus metabolites in the future.”
was changed to
“Firstly, several compounds produced by Bacillus spp. were listed. Then the isolation and identification techniques of Bacillus metabolites in recent years were described, including high-precision separation technology and omics technology for the separation of similar components of Bacillus metabolites. And the mechanisms of Bacillus metabolites against C. albicans were summarized from the inhibition of pathogenic fungi themselves and inhibition of the fungal virulence factors. The purpose of this review is to systematically summarize the recent studies on the inhibition of pathogenic fungi by Bacillus metabolites. The review is expected to become the reference for the control of pathogenic fungi such as C. albicans and the application of Bacillus metabolites in the future.”
Line 36: “detection” was changed to “prevalence”.
Line 47: “inhibit” was changed to “prevent”.
Line 54: “for the development of” was changed to “to develop”.
Line 65: “a kind of gram-positive” was changed to “Gram-positive”. “distributed” was changed to “existing”.
Line 80-85: “The lipopeptide biosurfactant produced by Indonesian marine bacteria, Bacillus subtilis C19 can inhibit the growth of C. albicans, which could be promoted as antifungal active agent [15]. The lipopeptide C16-fengycin A isolated from Bacillus amyloliquefaciens fmb60 showed significant antifungal activity against C. albicans [16].” was changed to “Indonesian marine bacteria Bacillus subtilis C19 could produce surfactin, which could inhibit the growth of C. albicans [16]. The lipopeptide C16-fengycin A isolated from Bacillus amyloliquefaciens fmb60 could produce C16-fengycin A, which showed significant anti-C. albicans activity [17].”
Line 91-95: “Since chitin and glucan are major components of fungal cell wall, lyases produced by Bacillus spp. are particularly effective against fungi [17]. Bacillus aryabhattai isolated from the Arabian Sea. B. aryabhattai can produce chitinases, which could convert chitin into chitin oligomers.” was changed to “Because chitin and glucan are the major components of fungal cell wall are chitin and glucan, lyases produced by Bacillus spp. are particularly effective against fungi [1718]. Bacillus aryabhattai isolated from the Arabian Sea. Bacillus aryabhattai isolated from the ocean can produce chitinases, which could convert chitin into chitin oligomers.”
Line 96-97: “against chitin powder” was removed.
Line 97-99: “The antifungal activity of chitinase against pathogenic fungi like C. albicans and Fusarium oxysporum revealed a zone of inhibition with 14 mm diameter [18].” was changed to “The antifungal activity of chitinase against pathogenic fungi such as C. albicans and Fusarium oxysporum exhibited inhibition zone with diameter of 14 mm [19].”
Line 100: “was” was removed. “.This strain produced” was changed to “could produce”.
Line 104-110: “Synergism activities of chitin-oligosaccharide and chitinase from this strain against fungi and pathogen Candida spp. (staining with methylene blue showed that almost 50% of 106 cells were died during 6 h) are promising for new anti-fungal drug with no side effect [19]. The molecular mechanisms underlying the inhibitory effects of Lactobacilli rhamnosus GG on hyphal morphogenesis were unraveled, which was a crucial step in C. albicans virulence.” was changed to “Chitin-oligosaccharide and chitinase had synergistic antifungal activity against Candida spp. They could kill 50% of 106 cells in 6 h and they were promising to be new antifungal agents without side effects [20]. The researchers revealed the molecular mechanisms of Lactobacilli rhamnosus GG inhibiting mycelial morphogenesis, which was a key step in the virulence of C. albicans.”
Line 111-118: “The activity of Msp1 was due to its ability to break down chitin, the main polymer in the hyphal cell wall of C. albicans [20]. Bacillus safensis physically attached and degraded rosary hyphae. Through genetic and phenotypic analysis, it was proved that the activity of bacterial chitinase to fungal cell wall chitin was a factor leading to the anti-pathogen effect of B. safensis [21].” was changed to “Msp1 exhibits antifungal activity due to its ability to degrade the major polymer chitin in the hyphal cell wall of C. albicans [21]. Bacillus safensis attached to C. albicans physically and degraded rosary hyphae. The activity of bacterial chitinase to fungal cell wall chitin was proved by genetic and phenotypic analysis to be the factor leading to the antifungal activity of B. safensis [22].”
Line 124-131: “A new Bacillus strain isolated from an Algerian salty lake can produce metabolites which were active against fungal pathogens. It was showed that the strain carried the gene clusters for the production of a number of bioactive and surface-active compounds. These included the lipopeptides surfactin and fengycin, antibacterial polyketides macrolactin and bacillaene, and a putative novel lanthipeptide [23].” was changed to “A new strain B. amyloliquefaciens isolated from a salty lake in Algeria could produce antifungal metabolites including the lipopeptides, the polyketones, and other new metabolites, which was proved by the gene clusters of the strain. The antifungal lipopeptides included surfactin and fengycin. The antibacterial polyketides included macrolactin and bacillaene, and the antifungal metabolite also included a putative novel lanthipeptide [24].”
Line 142: “to form a pore” was changed to “,by which a pore was formed”.
Line 144-145: “isolated from marine carpet clam, Paphia textile” was removed. Add “marine” before “Bacillus sp. Sh10”.
Line 178-179: “A new indole alkaloid was isolated from the deep-sea-derived bacterium Bacillus subterraneus 11593.” was changed to “The marine bacteria Bacillus subterraneus 11593 could produce a new indole alkaloid.”
Line 181-182: “to be (R)-N-[2-(3-hydroxy-2-oxoindolin-3-yl) ethyl] acetamide and named bacilsubteramide A” was removed.
Line 190: “(2:1 v/v)” was removed.
Line 191-192: Add “and” before “purified on”. “Further characterized and identified as” was changed to “Further structural confirmation indicated that it was”.
Line 194: “80% ammonium sulfate” was removed.
Line 195: “size-specific” and “using a Superdex-200 column” were removed.
Line 196-199: “After purification, the specific activity increased by 3.68 times, and the total activity recovery rate was 20.66%. The 11 kDa purity and molecular weight of the compound were determined by SDS-PAGE. The bacteriocin was analyzed by LC/MS/MS” was changed to “Through the purification process, the specific activity increased by 3.68 times, and the total activity recovery rate was 20.66%. The molecular weight of the compound was determined by SDS-PAGE. And the bacteriocin was analyzed by LC/MS/MS”.
Line 203-205: “, which was isolated by the slightly acidic (pH 5.82) Campanario hot spring (56.4 °C) in the Andes, central Chile. EPS” was removed.
Line 207-208: “analysis showed” was removed. Add “By the analysis of” and “, it was indicated”.
Line 213-217: “Researchers isolated a new Bacillus velezensis strain DTU001, studied its antifungal spectrum, sequenced its genome, and found the production of lipopeptide in HPLC-HRMS (HPLC-high resolution mass spectrometry) analysis. B. velezensis DTU001 exhibited excellent antiproliferative activity against C. albicans by producing multiple lipopeptides” was changed to “By the analysis of HPLC-HRMS (HPLC-high resolution mass spectrometry), the researchers found the lipopeptide produced by a new Bacillus velezensis strain DTU001. The antifungal activity of DTU001 was due to its ability to produce the lipopeptides that inhibit the proliferation of C. albicans”.
Line 234-237: “Genome mining revealed 19 candidate gene clusters encoding the biosynthesis of different secondary metabolites. A series of bacillibactins, fengycins, bacillomycins, surfactins, bacillaenes, and macrolactins were found by LC-DAD-MS with strong antibacterial activity” was changed to “The researchers first found candidate gene clusters that encode the biosynthesis of different secondary metabolites through genome mining, and further found a series of metabolites with strong antibacterial activity such as bacillibactins, fengycins, bacillomycins, surfactins, bacillaenes, and macrolactins by LC-DAD-MS”.
Line 237-238: “For the new Bacillus strain isolated from the Algerian Salt Lake, the strain” was changed to “The new Bacillus strain isolated from the salty lake”.
Line 242-244: “, including surfactin and fengycin, antibacterial polyketides macrolactin and bacillaene, and the putative new lanthipeptide. The ability of the strain to produce fengycin lipopeptides was confirmed by activity-guided purification using hydrophobic interaction chromatography” was changed to “Activity-directed purification by hydrophobic interaction chromatography confirmed the ability of the strain to produce fengycin lipopeptides”.
Line 247-248: “to analyze and identify” was changed to “for auxiliary analysis and identification of”.
Line 266: “destroy” was changed to “damage”. “integrity of the” and “cell” were removed.
Line 268: “resulting in” and cell were changed to “causing” and “cellular”.
Line 275: “can inhibit the growth of C. albicans” was changed to “had anti-C. albicans activity”.
Line 278: “The growth of C. albicans in AMP-17-treated cells” was changed to “After AMP-17 treatment, the growth of C. albicans”.
Line 284: “destroyed the cell wall integrity and” was changed to “can destroy the integrity of cell wall and the cell”.
Line 286-287: “resulting in rapid flow of ions including Ca2+” was changed to “which could cause rapid influx of Ca2+ or other ions”.
Line 292-294: “A soil bacterium B. safensis physically attached and degraded rosary hyphae. Through genetic and phenotypic analysis, it was proved that the activity of bacterial chitinase to fungal cell wall chitin was a factor leading to the anti-pathogen effect of B. safensis” was changed to “The bacterial chitinase produced by the soil bacterium B. safensis could destroy the fungal cell wall, which was a factor leading to the anti-pathogen effect of B. safensis”.
Line 301: “Ergosterol is an important component of fungal cell membrane” was changed to “An important component of fungal cell membrane is ergosterol”.
Line 302-303: “ergosterol-related genes” was changed to “genes related to ergosterol synthesis”.
Line 303-304: “The effect of surfactin on C. albicans was investigated by” was removed.
Line 304-305: Add “showed that surfactin can down-regulate the expression of”.
Line 307: “were down-regulated” and “surfactin decreased” were removed.
Line 310-311: “Surfactin could significantly affect physiology and gene transcription of C. albicans” was changed to “Surfactin exposure to C. albicans could cause physiological effects and affect gene transcription in C. albicans”.
Line 313-315: “the cell membrane ergosterol synthesis-related genes ERG1, ERG5, ERG6, and MET6 were down-regulated 5.88-, 17.54-, 13.33-, and 7.14-fold, respectively” was changed to “the expression of genes related to ergosterol synthesis (ERG1, ERG5, ERG6, and MET6) were down-regulated through genetic analysis”.
Line 317-321: “C16-Fengycin A had the ability to disrupt the cell wall due to its alterations of cell ultrastructure, and reduction of cell wall hydrophobicity. This was further confirmed by the changes in the exposure of the cell wall components and down-regulation of the genes related in the cell wall synthesis” was changed to “The changes of cell wall components exposure and the down-regulation of cell wall synthesis-related genes further proved that C16-fengycin A could destroy the cell wall of C. albicans, which was due to the fact that this lipopeptides could change the ultrastructure of cells and reduced the hydrophobicity of cell wall”.
Line 340-342: “The gene expression of ALS3, HWP1, BCR1, EFG1 and TEC1 was evaluated to investigate the effect of B. subtilis on C. albicans biofilm formation and filamentation” was changed to “By detecting the gene expression of ALS3, HWP1, BCR1, EFG1 and TEC1, the influence of B. subtilis on biofilm formation and hyphal formation of C. albicans was investigated”.
Line 345-346: “Among all the genes analyzed, ALS3 and HWP1 genes were the most affected” was changed to “ALS3 and HWP1 genes were most influenced of all the genes analyzed”.
Line 348-351: “B. subtilis reduces biofilm formation and filamentation of C. albicans by negatively regulating ALS3, HWP1, BCR1, EFG1 and TEC1 genes essential for biofilm and filamentation” was changed to “B. subtilis can down-regulate the expression of ALS3, HWP1, BCR1, EFG1 and TEC1 genes, which were necessary for biofilm formation and filamentation of C. albicans”.
Line 356-359: “B. safensis had anti-virulence factor activity against C. albicans, which strongly inhibited the formation of C. albicans filaments and biofilms” was changed to “By inhibiting the virulence factors of C. albicans, B. safensis had antifungal activity, which strongly inhibited the biofilm formation and filamentation of C. albicans”.
Line 363-364: “Lactobacillus paracasei isolate 28.4 reduced the filamentation of C. albicans in vitro” was changed to “Lactobacillus paracasei can reduce the in vitro filamentation of C. albicans”.
Line 365-366: “In vivo experiments also suggest that Lactobacillus paracasei 28.4 may be used as an alternative to control candidiasis” was removed.
Line 368: “Adhesion is the first step in fungal infection by C. albicans and” was changed to “Adhesion is the beginning of C. albicans infection”.
Line 370: “the adhesion of” was changed to “adhering”.
Line 372: “affect” was changed to “influence”.
Line 374-376: “The synergistic effect of the lipopeptide of B. subtilis AC7 combined with the group sensing molecule farnesol can counteract the C. albicans biofilm” was changed to “The lipopeptide of B. subtilis AC7 associated with farnesol (the group sensing molecule) can affect the C. albicans biofilm formation”.
Line 377: “inhibit” was changed to “prevent”.
Line 387: “mediate” was changed to “influence”. “to host cells” was removed.
Line 396: “virulence factors of C. albicans” was changed to “fungal virulence factors”.
Line 401: “drugs” was changed to “agents”.
Line 402: “production” was changed to “generation”.
Line 403: Add “(ROS)” after “reactive oxygen species”.
Line 405: “envelope” was changed to “biofilm”. “an important” was changed to ”a significant”.
Line 414: “reactive oxygen species” was removed.
Line 415-416: “With the treatment of C16-Fengycin A, the levels of ROS increased, resulting in mitochondrial dysfunction in the cells” was changed to “C16-Fengycin A can increase the levels of ROS, in which caused intracellular mitochondrial dysfunction”.
Line 418-419: “activate the High-Osmolarity Glycerol Mitogen-Activated Protein Kinase (HOG-MAPK) pathway and it could” was removed.
Line 427: “prevention and treatment” was changed to “control”.
Line 433-434: “C. albicans is” was changed to “As”. “which” was changed to “C. albicans”.
Point 2: English language correction is required, a few corrections are mentioned below:
Line 7: immunosuppressed instead of immunocompromised
line 34, in patients remove the
line 36: targets limit
line 51: remove "it was found that"
line 58: has "a" fast reproduction
LINE 60 : adaptability and a resistance to stress,....
line 135: have 'a' strong ability to inhibit
line 184: which can 'perform' efficient separation
line201: The researchers developed an integrated
line 204: and analysis of NMR data
line 220 : 'The' cell wall is the first target
line 226 : the improve fluconazole efficacy [42]
line 275: The hyphal state of C. albicans has a stronger ability for tissue invasion and infiltration.
Response 2: Thanks for your careful review. Some statements have been modified to improve originality, also added figures, so the position of some words has changed. The modifications listed below are in the lines of the revised version.
Line 7: “immunosuppressed” has been instead of “immunocompromised”. “immunocompromised” in line 32 was also modified.
line 41: “the” in “in the patients” has been removed.
line 43: “s” in “targets limits” has been removed.
line 58: "it was found that" has been removed.
line 66: “has fast reproduction” has been corrected to “has a fast reproduction”.
line 67 : “stress resistance” has been changed to “a resistance to stress”.
line 171: “have strong ability” has been corrected to “have a strong ability”.
line 226: The word “realize” has been changed to “perform”.
line 248: “Researchers” has been corrected to “The researchers”.
line 252: “NMR data analysis”has been corrected to “analysis of NMR data”.
line 269 : We added “The” before “cell wall is the first target”.
line 276 : “the efficacy of fluconazole” has been changed to “the improve fluconazole efficacy”.
line 334: “has stronger ability” has been corrected to “has a stronger ability”.
Author Response File: Author Response.docx
