Antibacterial activity on multidrug resistance bacteria stenotrophomonas maltophilia (strain k279a) and its correlation to secondary metabolic profile of heavy metal hyperaccumulator pteris vittata l

UNIVERSITY OF SCIENCE AND TECHNOLOGY OF HANOI UNIVERSITÉ DES SCIENCES ET DES TECHNOLOGIES DE HANOI UNDERGRADUATE SCHOOL FINAL REPORT BACHELOR GRADUATION By Tô Hiền Minh Project: Antibacterial activity on multidrug resistance bacteria Stenotrophomonas maltophilia (strain K279a) and its correlation to secondary metabolic profile of heavy metal hyperaccumulator Pteris vittata L Supervisor: Dr NGUYỄN THỊ KIỀU OANH University of Science and Technology of Hanoi Hanoi, November 2020 Index of Contents Contents Acknowledgements List of Abbreviations List of Figures and Tables Abstract: Tóm tắt: I Introduction: 1.1 Global context: 1.2 Literary review: 1.3 Objective: II Materials and Methods: 2.1 Chemicals 2.2 Instruments 2.3 Biological materials 2.4 Preparation of plant extracts 2.5 Bacterial and growth conditions 2.6 Antibacterial activity determination 10 2.7 Metabolite profiling of the methanolic extracts of P vittata 10 2.8 Data analysis 10 III Results: 12 3.1 Antibacterial effect of Pteris Vittata roots and leave extracts on resistance strain Stenotrophomonas maltophilia K279a: 12 3.2 Correlation between antibacterial activity of 70 Pteris Vittata extracts and its metabolic profile: 17 IV Discussion: 21 4.1 Discrimination on the activity of 70 Pteris vittata roots and leave extracts on S.malophilia strain K279a: 21 4.2 Correlation analysis between metabolic profile and antibacterial activity of Pteris Vittata 23 V Conclusion: 25 VI Reference: 26 VII Appendices Acknowledgements Foremost, I would like to express my deepest gratitude to my supervisors Dr Nguyễn Thị Kiều Oanh and PhD student Nguyễn Ngọc Liên from Life Sciences Department for their countless support, encouragement and guidance throughout my 3month internship in the University of Science and Technology Hanoi I could not have broadened my research skills and biostatistics knowledge without the opportunity that they give me to join the project and fulfill my bachelor thesis Besides, I want to thank all of the other professors and lecturers in the Life Science Department who always inspire us with their enthusiasm and passion for science - the best role models for us to become in the future Moreover, I’m sincerely proud to spend my most wonderful years of education with all of my friends in the PMAB class of 2020 We have always tried hard and inspired each other to break our own limits Especially Phạm Anh Thư and Trần Phương Trang for always supporting and motivating me The final semester I had an opportunity to an exchange program in Italy thanks to our university with a wonderful international cooperation strategy Most importantly, all of my thankfulness goes to my family and loved ones, they have nurtured and educated me to become who I am today, who is not afraid to learn and always grateful for what is coming in the future List of Abbreviations P vittata S maltophilia TMP-SMX MRSA MDR FDA ROS ESBL UPLC-QqQ-MS PCA PLS PLS- DA TSB DMSO CFU Gen Group P Group I Pteris vittata L Stenotrophomonas maltopilia trimethoprim- sulfamethoxazole Methicillin-resistant Staphylococcus aureus Multidrug resistant Food and Drug Administration Reactive oxygen species Extended spectrum betalactamase Ultra Performance Liquid Chromatography coupled triple quadrupole mass spectrometry Principle component analysis Partial least square regression Partial least square regression- Discrimination analysis Tryptone soya broth Dimethyl sulfoxide Colony forming unit Gentamicin Group promoting Group Inhibiting List of Figures and Tables Figures Figure Microdilution testing of P vittata root and leave extract on the growth curve of S maltophilia strain K279a Figure The inhibition percentage of P vittata leave and roots extracts on the growth of S maltophilia strain K279a Figure Correlation circles of PLS regression between metabolites content (M) and the inhibition percentage of S maltophilia K279a growth by leave and root extracts (Y) Tables Table I Paired t-Test statistic result on the difference between leave and root extracts activity Table II Pearson correlation coefficient statistics measure the linear correlation between metabolite intensity (X1) and antibacterial activity of leave or root extracts (X2) Abstract: Pteris vittata L is a heavy metal hyperaccumulator plant abundant in Thai Nguyen mining ore of Vietnam, which can change its secondary metabolic profile for environmental adaptation, leading to the shift of its synergistic bacterial system Under the same heavy metal stress, multidrug resistance bacteria can express intrinsic efflux pumps for both metal and antibiotics extrudation Investigation on P vittata bioactivity is a potential approach in finding novel antimicrobial agents to cope with an emerging era of multidrug resistance bacteria The resistant bacteria Stenotrophomonas maltophilia strain K279a is an opportunistic nosocomial pathogen which can cause severe respiratory tract infection Microdilution testing shows that P vittata roots and leave extracts have either antibacterial activity or growth promoting activity on the strain In our study, the first time metabolic profile of P vittata identified by a UPLC-QqQ-MS was statistically predicted for its correlation with antibacterial activity Flavonoid compounds such as pelargonin, quercetin were predicted to act against the S maltophilia and some alkaloids show its growth stimulator role These results bring out new directions to develop antibacterial lead compounds for treating clinical infection of Stenotrophomonas maltophilia K279a as well as to study its new resistance tendency Keywords: Pteris vittata, Stenotrophomonas maltophilia, antibacterial activity, bacterial growth promoting, secondary metabolites, PLS regression model Tóm tắt: Pteris vittata L thực vật hấp thụ kim loại nặng xuất nhiều vùng mỏ Thái Nguyên, Việt Nam Nó thay đổi chất chuyển hóa thứ cấp để thích nghi với mơi trường, dẫn đến thay đổi hệ vi khuẩn sống cộng sinh với Trong mơi trường kim loại nặng, vi khuẩn đa kháng thuốc hình thành bơm tống có chức đào thải kim loại nặng thuốc kháng sinh Việc nghiên cứu hoạt tính sinh học P vittata cách tiếp cận tiềm để tìm chất kháng khuẩn đối phó với thời đại vi khuẩn đa kháng thuốc lên Vi khuẩn đa kháng thuốc Stenotrophomonas maltophilia chủng K279a mầm bệnh hội bệnh viện gây nhiễm trùng đường hơ hấp nặng Thử nghiệm pha loãng đa nồng độ cho thấy dịch chiết từ rễ P vittata có hoạt tính kháng khuẩn hoạt tính thúc đẩy tăng trưởng chủng Trong nghiên cứu chúng tôi, lần chất chuyển hóa P vittata phân lập UPLC-QqQ-MS dự đoán thống kê mối tương quan với hoạt tính kháng khuẩn Các hợp chất flavonoid pelargonidin, quercetin dự đoán có tác dụng chống lại S maltophilia số alkaloid cho thấy vai trị kích thích tăng trưởng chủng Những kết mở hướng việc phát triển hợp chất kháng khuẩn để điều trị nhiễm trùng lâm sàng S maltophilia K279a nghiên cứu xu hướng kháng thuốc chủng vi khuẩn Từ khóa: Pteris vittata, Stenotrophomonas maltophilia, hoạt tính kháng khuẩn, hoạt tính tăng trưởng, thành phần chất chuyển hóa, hồi quy bình phương tối thiểu I Introduction 1.1 Global context: Antibiotic resistance situation worldwide Antibiotic resistance is becoming an urgent public health concern worldwide Human opportunistic pathogens cause millions of people with related diseases and thousands of deaths each year While becoming a burden for drug investigating and postsurgery infections prevention, the resistance strains show no sign of weakening (CDC, 2020) Reason for antibiotic resistance is not only the rapid evolving of bacteria themselves but also from anthropogenic activity: over-prescribing of antibiotics, excessive antibiotics used in agriculture and poor hygiene leading to nosocomial infections in hospital environments Others worth mentioning are the lack of rapid laboratory tests and global record of bacteria resistance situations In general, the most common nosocomial resistant bacteria strains are Pseudomonas aeruginosa, Stenotrophomonas maltophilia, Klebsiella pneumoniae, Staphylococcus aureus, Streptococcus pneumonia and Mycobacterium tuberculosis Report in 2018 shows approximately 640.000 cases of multidrug-resistant tuberculosis disease While penicillin as antibiotic against gram-positive bacteria experienced a trend of weakening in the ability to treat pneumonia, S maltophilia associated with respiratory tract infections also counters against its commonly used inhibitor trimethoprim- sulfamethoxazole (TMP-SMX)(“WHO | High Levels of Antibiotic Resistance Found Worldwide, New Data Shows” n.d.) In a worse fact, some strains become multi-resistance to any available clinical antimicrobial agents such as Methicillin-resistant Staphylococcus aureus(MRSA) which is notorious for hospital-acquired infections and resistant to methicillin, aminoglycosides, macrolides, tetracycline, vancomycin along with disinfectants Similarly, the occurrence of panresistant bacteria such as Pseudomonas aeruginosa, Acinetobacter baumannii, S maltophilia which conquers all of the antibiotics in standard panels via the mechanism of multidrug efflux pumps (Nikaido 2009) Recently, Covid-19 situation has also aggravated antibiotic resistance Statistics show that 50% of Covid-19 mortalities suffered from secondary infections, caused by multidrug resistance microorganism (bacterial or fungal) or coinfections Considering that these patients have already had underlined bacterial or fungal risk diseases (e.g: corticoid therapy, chronic respiratory disease, immune inflammatory response) and responded to a much higher rate of antibiotic usages when they are hospitalized (Rossato, Negrão, and Simionatto 2020) Plant as a novel source for producing antibiotics The bacteria possess resistance to multiple drug thanks to either one of these two mechanisms: (i) bacteria accumulate multiple genes, each gene encodes for resistance to a single drug within a single cell, occur on resistance plasmids, (ii) bacteria increase expression of genes which code for multidrug efflux pumps extruding a wide range of drugs (Nikaido 2009) From the first day, antibiotics were identified by Alexander Flemming as an accident leading to Penicillin invention Accordingly, the common sources of antibiotics come from secondary metabolites of fungi or soil bacteria produced by these industrial methods: fermentation (e.g: beta lactam, penicillins, cephalosporins); semi-synthetic; synthetic pathway In this scenario, the main source of antibiotics - soil actinomycetes has been overmined and the investigation based on this source is said to be useless due to the high rediscovery rate of known compounds (Lewis 2017) Rising as a novel candidate to conquer MDR bacteria, plant resources have long been used as herbal medicine to treat a vast array of inflammatory and bacterial infections It’s reported through archeology that in the primitive era, humans benefited partially from plant and metal to treat microbial infections: Neanderthals living 60,000 years ago in present-day Iraq used plants such as hollyhock to treat infections Since plants can produce a variety of primary and secondary metabolites, these compounds are not only active to defend insects, microorganisms, herbivores and plant pathogens but also against human pathogens The major groups are: phenolics, polyphenols, terpenoids, essential oils, lectins and polypeptides, alkaloids or polyacetylenes They can act solely or in combination with different compounds by a variety of mechanisms: protein inactivation, adhesins and enzymes, block cell-to-cell signaling, quench production of virulence factors such as exotoxin, disrupt or inhibit formations of biofilms, shielding of pathogens during an infection (Cowan 1999) Despite such potentials, antimicrobial drugs from plants only consist of 3% drug approved by FDA Thus, the current investigation on antimicrobial potential of plants is merely the tip of the iceberg Pteris vittata L antibacterial potential Pteris vittata L., also known as Chinese Brake Fern, is well studied for its metal hyperaccumulator function found in anthropogenic polluted ore mining sites in Thai Nguyen province of Vietnam A number of previous publications focus on its phytoremediation role, especially for the individual absorption of Pb, Zn, As and their combination For Arsenic, it can absorb up to 23,000 micro gram arsenic g-1 in its frond or aerial parts, 4-8 folds more efficiently than soil extraction method (Alkorta et al 2004) Otherwise, due to the phytotoxic correlation of excessive heavy metal and production of , P vittata is proved to have antioxidant activity by scavenging toxic free radicals and ROS’s enzymatic or non enzymatic defense mechanism However, there is a limit of papers investigating the antibacterial effect of P vittata or considering its roots and bacteria synergism affected by the secondary metabolites While considering secondary metabolites as strong antioxidants to effectively protect against oxidative stress, we should also put eyes on the antibacterial potential of the diverse metabolic profile of P vittata Appendix Curve S3, S4 Q2 RMSEP (Root mean square error of prediction), components choosing for leave (left) and roots extracts (right) Appendix Curve S5, S6, cumulative r2, q2 of leave (left) and root (right) extracts Plot S7 Predictive power validation of PLS model for leave extracts, components Plot S8 Predictive power validation of PLS model for root extracts, components Table S9 359 Metabolites identified by UPLC-QqQ-MS targeted metabolomic approach M Annotation Group Type of metabolite 4-Hydroxy-3-methoxycinnamaldehyde Phenylpropanoic acids and derivatives Secondary metabolite N-6-(delta-2-Isopentenyl) adenosine Nucleosides, nucleotides, and analogues Primary metabolite Shikimic acid Carboxylic acids and derivatives Primary metabolite Phenylephrine Phenols Secondary metabolite 1,1-Dimethylbiguanide Carboxylic acids and derivatives Primary metabolite 1-Aminocyclopropane-1-carboxylic acid Carboxylic acids and derivatives Primary metabolite Serotonin Alkaloids and derivatives Secondary metabolite 5-Methylcytosine Other Primary metabolite 6-(gamma,gamma-Dimethylallylamino)purine Other Primary metabolite 10 Adenosine Nucleosides, nucleotides, and analogues Primary metabolite 11 Anthranilic acid Benzene and substituted derivatives Secondary metabolite 12 Caffeine Alkaloids and derivatives Secondary metabolite 12 Caffeine Alkaloids and derivatives Secondary metabolite 13 cis-Aconitic Acid Carboxylic acids and derivatives Primary metabolite 14 Glycine Carboxylic acids and derivatives Primary metabolite 15 Histidine Carboxylic acids and derivatives Primary metabolite 16 Methionine Carboxylic acids and derivatives Primary metabolite 17 Phenylalanine Carboxylic acids and derivatives Primary metabolite 18 Tryptophane Carboxylic acids and derivatives Primary metabolite 19 Tyrosine Carboxylic acids and derivatives Primary metabolite 20 Sucrose Sugar Primary metabolite 21 Uracil Nucleosides, nucleotides, and analogues Primary metabolite 22 Kynurenine Other Primary metabolite 23 3-(2-Aminoethyl)indole Alkaloids and derivatives Secondary metabolite 24 Galacturonic acid Sugar Primary metabolite 25 2-Hydroxypyridine Organoheterocyclic compounds Primary metabolite 26 Mesaconic acid Carboxylic acids and derivatives Primary metabolite 27 Methylmalonic acid Carboxylic acids and derivatives Primary metabolite 28 Pyridoxamine Organoheterocyclic compounds Primary metabolite 29 3-Methyladenine Nucleosides, nucleotides, and analogues Primary metabolite 30 4-Aminobenzoic acid Benzene and substituted derivatives Secondary metabolite 31 Agmatine Organonitrogen compounds Primary metabolite 32 Glucosamine-6-phosphate Sugar Primary metabolite 33 Harmaline Alkaloids and derivatives Secondary metabolite 34 Carnosine Carboxylic acids and derivatives Primary metabolite 35 Leupeptin Carboxylic acids and derivatives Primary metabolite 36 Pterine Organonitrogen compounds Primary metabolite 37 1-Methylhistamine Alkaloids and derivatives Secondary metabolite 38 2-Aminoethylphosphonic acid Phosphate Primary metabolite 39 2'-Deoxyadenosine Nucleosides, nucleotides, and analogues Primary metabolite 40 2'-Deoxycytidine Nucleosides, nucleotides, and analogues Primary metabolite 41 2'-Deoxyguanosine Nucleosides, nucleotides, and analogues Primary metabolite 42 2'-Deoxyinosine Organonitrogen compounds Primary metabolite 43 3,4-Dihydroxy-phenylalanine Carboxylic acids and derivatives Primary metabolite 44 3-Amino-1,2,4-triazole Organoheterocyclic compounds Primary metabolite 45 Adenosine-3',5'-cyclicmonophosphate Nucleosides, nucleotides, and analogues Primary metabolite 46 Adonitol Sugar Primary metabolite 47 Allantoin Organoheterocyclic compounds Primary metabolite 48 Amantadine Alkaloids and derivatives Secondary metabolite 49 Argininosuccinic acid Carboxylic acids and derivatives Primary metabolite 50 Benzamidine Organoheterocyclic compounds Primary metabolite 51 Benzocaine Carboxylic acids and derivatives Primary metabolite 52 Caffeic acid Phenylpropanoic acids and derivatives Secondary metabolite 53 Creatine Carboxylic acids and derivatives Primary metabolite 54 Creatinine Carboxylic acids and derivatives Primary metabolite 55 Cystathionine Carboxylic acids and derivatives Primary metabolite 56 Cytidine Nucleosides, nucleotides, and analogues Primary metabolite 57 Cytidine-3',5'-cyclicmonophosphate Nucleosides, nucleotides, and analogues Primary metabolite 58 Diethanolamine Organoheterocyclic compounds Primary metabolite 59 2,3-Diaminopropionic acid Carboxylic acids and derivatives Primary metabolite 60 Hydroxybutyric acid Carboxylic acids and derivatives Primary metabolite 61 Ribose-5-phosphate Sugar Primary metabolite 62 Riboflavin-5'-monophosphate Nucleosides, nucleotides, and analogues Primary metabolite 63 Fusaric acid Carboxylic acids and derivatives Primary metabolite 64 Gibberellin A4 Terpenoids Secondary metabolite 65 Glutathione (oxidized form) Carboxylic acids and derivatives Primary metabolite 66 Gly-Gly Carboxylic acids and derivatives Primary metabolite 67 Guanine Organoheterocyclic compounds Primary metabolite 68 Guanosine Nucleosides, nucleotides, and analogues Primary metabolite 69 Guanosine-3',5'-cyclic monophosphate Nucleosides, nucleotides, and analogues Primary metabolite 70 Hypotaurine Organosulfur compounds Primary metabolite 71 3-Indoleacetic acid Alkaloids and derivatives Secondary metabolite 72 Inosine Nucleosides, nucleotides, and analogues Primary metabolite 73 Inosine-5'-monophosphate Nucleosides, nucleotides, and analogues Primary metabolite 74 Isoguvacine Other Primary metabolite 75 Asparagine Carboxylic acids and derivatives Primary metabolite 76 Aspartic acid Carboxylic acids and derivatives Primary metabolite 77 Cystine Carboxylic acids and derivatives Primary metabolite 78 Leucylleucyltyrosine Carboxylic acids and derivatives Primary metabolite 79 Glutamic acid Carboxylic acids and derivatives Primary metabolite 80 Glutamine Carboxylic acids and derivatives Primary metabolite 81 Glutathione (reduced form) Carboxylic acids and derivatives Primary metabolite 82 Histidinol Carboxylic acids and derivatives Primary metabolite 83 Homocarnosine Carboxylic acids and derivatives Primary metabolite 84 Homocysteine Carboxylic acids and derivatives Primary metabolite 85 Ornithine Carboxylic acids and derivatives Primary metabolite 86 Proline Carboxylic acids and derivatives Primary metabolite 87 Pyroglutamic acid Carboxylic acids and derivatives Primary metabolite 88 Serine Carboxylic acids and derivatives Primary metabolite 89 Maltotriose Sugar Primary metabolite 90 Mecamylamine Other Primary metabolite 91 1-Methylguanidine Other Primary metabolite 92 3-Ureidopropionic acid Other Primary metabolite 93 N-Formyl-methionine Carboxylic acids and derivatives Primary metabolite 94 Nicotinic acid mono nucleotide Organoheterocyclic compounds Primary metabolite 95 2-Hydroxycinnamic acid Phenylpropanoic acids and derivatives Secondary metabolite 96 Orotic acid Organoheterocyclic compounds Primary metabolite 97 Piperacillin Other Primary metabolite 98 4-Nitrophenol Phenols Secondary metabolite 99 3,4-Dihydroxybenzoic acid Benzene and substituted derivatives Secondary metabolite 100 Pyridoxine Organoheterocyclic compounds Primary metabolite 101 Quisqualic acid Carboxylic acids and derivatives Primary metabolite 102 S-Carboxymethyl-L-cysteine Carboxylic acids and derivatives Primary metabolite 103 Sebacic acid Carboxylic acids and derivatives Primary metabolite 104 Spermine Alkaloids and derivatives Secondary metabolite 105 Succinic acid Carboxylic acids and derivatives Primary metabolite 106 Thymidine Nucleosides, nucleotides, and analogues Primary metabolite 107 Thymidine-3',5'-cyclic monophosphate Nucleosides, nucleotides, and analogues Primary metabolite 108 Thymine Organoheterocyclic compounds Primary metabolite 109 trans-Zeatin-riboside Nucleosides, nucleotides, and analogues Primary metabolite 110 Tyramine Organonitrogen compounds Primary metabolite 111 Uridine-5'-monophosphate Nucleosides, nucleotides, and analogues Primary metabolite 112 alpha-Lactose Sugar Primary metabolite 113 3-Guanidinopropionic acid Nucleosides, nucleotides, and analogues Primary metabolite 114 beta-Nicotinamide mononucleotide Other Primary metabolite 115 Phenylpyruvate Carboxylic acids and derivatives Primary metabolite 116 gamma-Amino-n-butyric acid Carboxylic acids and derivatives Primary metabolite 117 delta-Aminolevulinic acid Carboxylic acids and derivatives Primary metabolite 118 6-Aminohexanoic acid Carboxylic acids and derivatives Primary metabolite 119 Catechol Phenols Secondary metabolite 120 Eriodictyol Flavonoids Secondary metabolite 121 Genistein Isoflavonoids Secondary metabolite 122 Naringenin Flavonoids Secondary metabolite 123 Pelargonin Flavonoids Secondary metabolite 124 Apigenin glu Flavonoids Secondary metabolite 125 Rhoifolin Flavonoids Secondary metabolite 126 Keracyanin Flavonoids Secondary metabolite 127 Isorhamnetin glu Flavonoids Secondary metabolite 128 Robinin Flavonoids Secondary metabolite 129 Kaempferol glu Flavonoids Secondary metabolite 130 Homoorietin Flavonoids Secondary metabolite 132 Puerarin Isoflavonoids Secondary metabolite 133 Peltatoside Flavonoids Secondary metabolite 134 Hyperoside Flavonoids Secondary metabolite 135 Quercetin-3-O-glu-6''-acetate Flavonoids Secondary metabolite 136 Capsaicin Phenols Secondary metabolite 137 Dihydrocapsaicin Phenols Secondary metabolite 138 Nordihydrocapsaicin Phenols Secondary metabolite 139 4-Coumaric acid Phenylpropanoic acids and derivatives Secondary metabolite 140 Rosmarinic acid Phenylpropanoic acids and derivatives Secondary metabolite 141 Kaempferol-7-O-alpha-rham Flavonoids Secondary metabolite 142 3-Cyanopyridine Organoheterocyclic compounds Primary metabolite 143 Urocanic acid Organoheterocyclic compounds Primary metabolite 144 N-Acetylneuraminic acid Other Primary metabolite 145 N-Acetyl-mannosamine Sugar Primary metabolite 146 O-Acetyl-serine Carboxylic acids and derivatives Primary metabolite 147 4-Methylumbelliferyl acetate Coumarins and derivatives Secondary metabolite 148 Pyridoxal-5'-phosphate Organoheterocyclic compounds Primary metabolite 149 Pyridoxal Organoheterocyclic compounds Primary metabolite 150 N-Acetyl putrescine Alkaloids and derivatives Secondary metabolite 151 Arabitol Sugar Primary metabolite 152 Glucoheptose Sugar Primary metabolite 153 3-Hydroxyanthranilic acid Benzene and substituted derivatives Secondary metabolite 154 Jasmonic acid Carboxylic acids and derivatives Primary metabolite 155 Abscisic acid Terpenoids Secondary metabolite 156 beta-Homovaline Carboxylic acids and derivatives Primary metabolite 157 Quinic acid Carboxylic acids and derivatives Primary metabolite 158 Dihydroorotic acid Other Primary metabolite 159 Chlorogenic acid Phenylpropanoic acids and derivatives Secondary metabolite 160 6-Hydroxynicotinic Acid Organoheterocyclic compounds Primary metabolite 161 O-Succinyl-homoserine Carboxylic acids and derivatives Primary metabolite 162 Gibberelin A3 Terpenoids Secondary metabolite 163 cis,cis-Muconic acid Carboxylic acids and derivatives Primary metabolite 164 Pimelic acid Carboxylic acids and derivatives Primary metabolite 165 4-Methyl-5-thiazoleethanol Organoheterocyclic compounds Primary metabolite 166 O-Acetyl-homoserine Carboxylic acids and derivatives Primary metabolite 167 Ala-ala Carboxylic acids and derivatives Primary metabolite 168 Carbamoyl-aspartic acid Carboxylic acids and derivatives Primary metabolite 169 5'-Deoxy-5'-Methylthioadenosine Nucleosides, nucleotides, and analogues Primary metabolite 170 Dihydrozeatin Nucleosides, nucleotides, and analogues Primary metabolite 171 2-Isopropylmalic acid Carboxylic acids and derivatives Primary metabolite 172 3-Methyl-2-oxobutyric acid Carboxylic acids and derivatives Primary metabolite 173 Scoulerin Alkaloids and derivatives Secondary metabolite 174 allo-Threonine Carboxylic acids and derivatives Primary metabolite 175 Taxifolin Flavonoids Secondary metabolite 176 Lignoceric Acid Carboxylic acids and derivatives Primary metabolite 177 Methyl Jasmonate Carboxylic acids and derivatives Primary metabolite 178 Vanillin Benzene and substituted derivatives Secondary metabolite 179 4-Hydroxypyridine Organoheterocyclic compounds Primary metabolite 180 gamma-Glu-Cys Carboxylic acids and derivatives Primary metabolite 181 erythro-Dihydrosphingosine Lipids and lipid-like molecules Primary metabolite 182 Zeatin-9-glucoside Other Primary metabolite 183 Syringaldehyde Benzene and substituted derivatives Secondary metabolite 184 Citraconic Acid Carboxylic acids and derivatives Primary metabolite 185 5-Aminovaleric acid Carboxylic acids and derivatives Primary metabolite 186 3,4-Dimethoxycinnamic acid Phenylpropanoic acids and derivatives Secondary metabolite 187 3,5-Dimethoxycinnamic acid (predominantly trans) Phenylpropanoic acids and derivatives Secondary metabolite 188 m-Hydroxycinnamic acid Phenylpropanoic acids and derivatives Secondary metabolite 189 3-Hydroxypyridine Organoheterocyclic compounds Primary metabolite 190 4-Methoxycinnamic acid Phenylpropanoic acids and derivatives Secondary metabolite 191 Hesperetin Flavonoids Secondary metabolite 192 Itaconic acid Carboxylic acids and derivatives Primary metabolite 193 N-Acetylglycine Carboxylic acids and derivatives Primary metabolite 194 trans-3,5-Dimethoxy-4hydroxycinnamaldehyde Phenylpropanoic acids and derivatives Secondary metabolite 195 Sorbitol-6-phosphate Sugar Primary metabolite 196 Suberic acid Carboxylic acids and derivatives Primary metabolite 197 Threonic acid Sugar Primary metabolite 198 2,2',2''-Nitrilotriethanol Organonitrogen compounds Primary metabolite 199 Phosphocholine,1-Lauroyl-2-Hydroxy-snGlycero-3- Lipids and lipid-like molecules Primary metabolite 200 Phosphocholine,1-Decanoyl-2-Hydroxy-snGlycero-3- Lipids and lipid-like molecules Primary metabolite 201 Glucosyl-beta-1-1'-erythoro-Sphingosine Lipids and lipid-like molecules Primary metabolite 202 4-Deoxyphloridzin Flavonoids Secondary metabolite 203 2',6'-Dihydroxy-4-Methoxychalcone-4'-O-neo Flavonoids Secondary metabolite 204 Flavanomarein Flavonoids Secondary metabolite 205 Hesperidin Flavonoids Secondary metabolite 206 Luteolin-3',7-di-O-glu Flavonoids Secondary metabolite 207 Luteolin-4'-O-glu Flavonoids Secondary metabolite 208 Neoeriocitrin Flavonoids Secondary metabolite 209 Neohesperidin Flavonoids Secondary metabolite 210 Neohesperidin dihydrochalcone Flavonoids Secondary metabolite 211 Phloridzin Flavonoids Secondary metabolite 212 Poncirin Flavonoids Secondary metabolite 213 Saponarin Flavonoids Secondary metabolite 214 Sissotrin Isoflavonoids Secondary metabolite 215 Solasodine Alkaloids and derivatives Secondary metabolite 216 Vitexin Flavonoids Secondary metabolite 217 Methyl dihydrojasmonate Carboxylic acids and derivatives Primary metabolite 218 Acetylserine Carboxylic acids and derivatives Primary metabolite 219 S-(5'-Adenosyl)-methionine Nucleosides, nucleotides, and analogues Primary metabolite 220 Kaempferol trihex Flavonoids Secondary metabolite 221 Kaempferol ara Flavonoids Secondary metabolite 222 Kaempferol gluc Flavonoids Secondary metabolite 223 Myricetin gal Flavonoids Secondary metabolite 224 Myricetin xyl Flavonoids Secondary metabolite 225 Quercetin-3-Gluc Flavonoids Secondary metabolite 226 alpha-Methyl-histidine Carboxylic acids and derivatives Primary metabolite 227 3-Hydroxy-3-methylglutarate Carboxylic acids and derivatives Primary metabolite 228 3-Indoxylsulfate Alkaloids and derivatives Secondary metabolite 229 2,5-Dihydroxy benzoic acid Benzene and substituted derivatives Secondary metabolite 230 Hippuric Acid Benzene and substituted derivatives Secondary metabolite 231 Salicylic Acid Benzene and substituted derivatives Secondary metabolite 232 Trimethylamine N-oxide Organoheterocyclic compounds Primary metabolite 233 3-Methylhistidine Carboxylic acids and derivatives Primary metabolite 234 5-Hydroxyindole-3-acetate Carboxylic acids and derivatives Primary metabolite 235 5-Hydroxylysine Carboxylic acids and derivatives Primary metabolite 236 p-Hydroxyphenyllactic acid Phenylpropanoic acids and derivatives Secondary metabolite 237 2-Hydroxyisocaproic acid Carboxylic acids and derivatives Primary metabolite 238 3-Phenyllactic acid Phenylpropanoic acids and derivatives Secondary metabolite 239 Homovanillic acid Phenols Secondary metabolite 240 Sulfocysteine Carboxylic acids and derivatives Primary metabolite 241 N-Acetyl-aspartic acid Carboxylic acids and derivatives Primary metabolite 242 Glycyl-proline Carboxylic acids and derivatives Primary metabolite 243 N-Tigloylglycine Carboxylic acids and derivatives Primary metabolite 244 Riboflavin Nucleosides, nucleotides, and analogues Primary metabolite 245 Oxypurinol Organoheterocyclic compounds Primary metabolite 246 trans-Aconitic acid Carboxylic acids and derivatives Primary metabolite 247 3-Hydroxy-3-methylbutanoic acid Carboxylic acids and derivatives Primary metabolite 248 N-Isovaleroylglycine Carboxylic acids and derivatives Primary metabolite 249 Choline Organonitrogen compounds Primary metabolite 250 Kynurenic acid Carboxylic acids and derivatives Primary metabolite 251 Acetaminophen Phenols Secondary metabolite 252 Glycocyamine Carboxylic acids and derivatives Primary metabolite 253 Trigonelline Alkaloids and derivatives Secondary metabolite 254 Phenaceturic acid Carboxylic acids and derivatives Primary metabolite 255 1,3-Dimethylurate Alkaloids and derivatives Secondary metabolite 256 3-Methylxanthine Alkaloids and derivatives Secondary metabolite 257 3-Hydroxymandelic acid Phenols Secondary metabolite 258 4-Hydroxy-3-methoxybenzoic acid Benzene and substituted derivatives Secondary metabolite 259 4-Pyridoxate Organoheterocyclic compounds Primary metabolite 260 5-Methoxysalicylic acid Benzene and substituted derivatives Secondary metabolite 261 N-Acetyl-glutamic acid Carboxylic acids and derivatives Primary metabolite 262 3,5-Dibromo-tyrosine Carboxylic acids and derivatives Primary metabolite 263 1-Isothiocyanato-7-(methylsulfinyl)-heptane Organosulfur compounds Primary metabolite 264 1-Isothiocyanato-6-(methylsulfinyl)-hexane Organosulfur compounds Primary metabolite 265 1-Isothiocyanato-9-(methylsulfinyl)-nonane Organosulfur compounds Primary metabolite 266 1-Isothiocyanato-8-(methylsulfinyl)-octane Organosulfur compounds Primary metabolite 267 1-Isothiocyanato-6-(methylsulfonyl)-hexane Organosulfur compounds Primary metabolite 268 Sulforaphene Organosulfur compounds Primary metabolite 269 Indole-3-carboxylic acid Alkaloids and derivatives Secondary metabolite 270 Maleic acid Carboxylic acids and derivatives Primary metabolite 271 Cyanidin-3-O-rham Flavonoids Secondary metabolite 272 Zearalenone Other Primary metabolite 273 Glucuronic acid Sugar Primary metabolite 274 Betaine Alkaloids and derivatives Secondary metabolite 275 Quercetin trihex Flavonoids Secondary metabolite 276 1-O-b-glucopyranosyl sinapate Phenylpropanoic acids and derivatives Secondary metabolite 277 Glyceric acid Carboxylic acids and derivatives Primary metabolite 278 Folic acid Other Primary metabolite 279 Procyanidin C1 Flavonoids Secondary metabolite 280 Prostaglandin E1 Carboxylic acids and derivatives Primary metabolite 281 Cyanidin ara Flavonoids Secondary metabolite 282 Peonidin ara Flavonoids Secondary metabolite 283 Petunidin glu Flavonoids Secondary metabolite 284 Quercetin dihex Flavonoids Secondary metabolite 285 Cyanidin dihex Flavonoids Secondary metabolite 286 Peonidin dihex Flavonoids Secondary metabolite 287 Delphinidin dihex Flavonoids Secondary metabolite 288 Cyanidin trihex Flavonoids Secondary metabolite 289 Cyanidin coum trihex Flavonoids Secondary metabolite 290 Petunidin coum trihex Flavonoids Secondary metabolite 291 E-3,4,5'-Trihydroxy-3'glucopyranosylstilbene Stilbenoids Secondary metabolite 292 E-4,5'-Dihydroxy-3-methoxy-3'glucopyranosylstilbene Stilbenoids Secondary metabolite 293 Ethionine Carboxylic acids and derivatives Primary metabolite 294 o-Anisic Acid Benzene and substituted derivatives Secondary metabolite 295 Sodium deoxycholate Other Primary metabolite 296 Daphnetin Coumarins and derivatives Secondary metabolite 297 6,7-Dihydroxycoumarin Coumarins and derivatives Secondary metabolite 298 Esculin sesquihydrate Coumarins and derivatives Secondary metabolite 299 Melatonin Alkaloids and derivatives Secondary metabolite 300 Kaempferol trihex Flavonoids Secondary metabolite 301 Isorhamnetin dihex Flavonoids Secondary metabolite 302 Kaempferol-3-O-rut Flavonoids Secondary metabolite 303 Acacetin dihex Flavonoids Secondary metabolite 304 Cyanidin dihex Flavonoids Secondary metabolite 305 Kaempferol dihex or Vitexin rham Flavonoids Secondary metabolite 306 Kaempferol rham Flavonoids Secondary metabolite 307 Cyanidin hex Flavonoids Secondary metabolite 308 Maritimein or Luteolin glu Flavonoids Secondary metabolite 309 Eriodictyol glu or Marein Flavonoids Secondary metabolite 310 Quercetin hex Flavonoids Secondary metabolite 311 Quercetin hex Flavonoids Secondary metabolite 312 Malvidin hex Flavonoids Secondary metabolite 313 Syringetin gal or Syringetin glu Flavonoids Secondary metabolite 314 Peonidin gal or Peonidin glu Flavonoids Secondary metabolite 315 Cytidine-5'-diphosphocholine Organoheterocyclic compounds Primary metabolite 316 Catechin Flavonoids Secondary metabolite 317 Sodium or Calcium pantothenate Organooxygen compounds Primary metabolite 318 alpha-Galactose-1-phosphate or alphaGlucose-1-phosphate Sugar Primary metabolite 319 Mannose-6-phosphate Sugar Primary metabolite 320 Hex-6-phosphate Sugar Primary metabolite 321 Galactosamine or Glucosamine Sugar Primary metabolite 322 or Methoxycinnamic acid Phenylpropanoic acids and derivatives Secondary metabolite 323 4-Methylumbelliferone Coumarins and derivatives Secondary metabolite 324 Theophylline or 1,7-Dimethylxanthine Alkaloids and derivatives Secondary metabolite 325 cis or trans Zeatin Lipids and lipid-like molecules Primary metabolite 326 Thioctamide Organosulfur compounds Primary metabolite 327 Ferulic acid Phenylpropanoic acids and derivatives Secondary metabolite 328 Guanosine-5'-monophosphate Nucleosides, nucleotides, and analogues Primary metabolite 329 Cellobiose Sugar Primary metabolite 330 Dihex Sugar Primary metabolite 331 Hexane-hexol Sugar Primary metabolite 332 Arginine or Acetyl ornithine or Citrulline Carboxylic acids and derivatives Primary metabolite 333 Glycerol phosphoate Lipids and lipid-like molecules Primary metabolite 334 Histamine or Cytosine Organoheterocyclic compounds Primary metabolite 335 Nicotinamide Organoheterocyclic compounds Primary metabolite 336 Glycolaldehyde dimer or tetrose Other Primary metabolite 337 Alanine or Sarcosine Carboxylic acids and derivatives Primary metabolite 338 1,3-Diaminopropane Other Primary metabolite 339 Norvaline or Valine Carboxylic acids and derivatives Primary metabolite 340 Threonine or Methylserine or Homoserine Carboxylic acids and derivatives Primary metabolite 341 2-Hydroxyisobutyric acid Carboxylic acids and derivatives Primary metabolite 342 2-Aminobutyric acid or N,N-Dimethylglycine or N-Methyl alanine Carboxylic acids and derivatives Primary metabolite 343 2-Hydroxyisovaleric acid Carboxylic acids and derivatives Primary metabolite 344 Indole-3-carboxyaldehyde Alkaloids and derivatives Secondary metabolite 345 Malic acid Carboxylic acids and derivatives Primary metabolite 346 alpha-Ketoglutaric acid Carboxylic acids and derivatives Primary metabolite 347 Mandelic acid Benzene and substituted derivatives Secondary metabolite 348 Carnitine Other Primary metabolite 349 2-Aminoadipic Acid Carboxylic acids and derivatives Primary metabolite 350 Adipic acid or 2-Methylglutaric acid Carboxylic acids and derivatives Primary metabolite 351 L-Citramalic acid Carboxylic acids and derivatives Primary metabolite 352 4-Hydroxybenzoate Benzene and substituted derivatives Secondary metabolite 353 Ethylmalonic acid or Glutaric acid or Methylsuccinic acid Carboxylic acids and derivatives Primary metabolite 354 Lysine Carboxylic acids and derivatives Primary metabolite 355 Nicotinic acid or Isonicotinic acid Organoheterocyclic compounds Primary metabolite 356 Norleucine or Alloisoleucine Carboxylic acids and derivatives Primary metabolite 357 Quercitrin or Quercetin rham Flavonoids Secondary metabolite 358 Rutin Flavonoids Secondary metabolite 359 Melezitose or 1-Kestose Sugar Primary metabolite ... antibacterial activity of methanol extract of leave and roots of P vittata L against MDR S maltophilia (strain K279a) (ii) Correlation between metabolic profile and bacterial inhibition percentage and. .. heavy metals (Cao, Ma, and Tu 2004) Pteris vittata? ??s secondary metabolites and soil microbial community correlation in antibacterial activity: In the heavy metal polluted area, soil bacterial communities... via their secondary metabolites, can give an influence on the acquisition/expression of MDR bacteria, especially with S maltophilia (strain K279a) Metabolic profiling and correlation analysis by