title: Identification of antimicrobial compound, diketopiperazines, from a Bacillus sp. N strain associated with a rhabditid entomopathogenic nematode against major plant pathogenic fungi.
abstract: OBJECTIVE
  To purify and characterize antimicrobial compounds from Bacillus sp. strain N associated with rhabditid entomopathogenic nematode (EPN).
  
  
  RESULTS
  The cell-free culture filtrate of a bacterium associated with an EPN, Rhabditis (Oscheius) sp., exhibited strong antimicrobial activity. The ethyl acetate extract of the bacterial culture filtrate was purified by silica gel column chromatography to obtain three diketopiperazines (DKPs). The structure and absolute stereochemistry of this compound were determined based on extensive spectroscopic analyses (FABMS, (1) H NMR, (13) C NMR, (1) H-(1) H COSY, (1) H-(13) C HMBC) and Marfey's method. The compounds were identified as cyclo(l-Pro-l-Leu), cyclo(d-Pro-l-Leu) and cyclo(d-Pro-l-Tyr), respectively. Three DKPs were active against all the five fungi tested (Aspergillus flavus, Candida albicans, Fusarium oxysporum, Rhizoctonia solani and Penicillium expansum) and are more effective than the standard fungicide bavistin. The highest activity of 4 μg ml(-1)  by cyclo(l-Pro-l-Leu) and cyclo(d-Pro-l-Tyr) was recorded against P. expansum, a plant pathogen responsible for causing postharvest decay of stored apples and oranges. Cyclo(d-Pro-l-Leu) recorded good antibacterial activity against all the four bacteria tested (Bacillus subtilis, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa), and cyclo(l-Pro-l-Leu) and cyclo(d-Pro-l-Tyr) recorded good activity only against Gram-positive bacteria. To our knowledge, this is the first report of antifungal activity of the DKPs against the plant pathogenic fungi F. oxysporum, R. solani and P. expansum. The production of cyclo(l-Pro-l-Leu), cyclo(d-Pro-l-Leu) and cyclo-(d-Pro-l-Tyr) by a bacterium associated with EPN is also reported here for the first time.
  
  
  CONCLUSIONS
  Isolated DKPs demonstrated high antimicrobial activity against bacteria and fungi, especially against plant pathogenic fungi. We conclude that the bacterium associated with EPN is a promising source of natural bioactive secondary metabolites which may receive great benefit in the field of agriculture.
  
  
  CONCLUSIONS
  This study is a significant contribution to the knowledge of compounds unique from EPN bacteria as potential sources of new drugs in the agricultural and pharmacological industry.
authors: Division of Crop Protection/Division of Crop Utilisation, Central Tuber Crops Research Institute, Sreekariyam, Thiruvananthapuram, India. nishanthctcri@gmail.com
authors: S
authors: S
authors: Nishanth Kumar
authors: C
authors: C
authors: Mohandas
authors: J V
authors: JV
authors: Siji
authors: K N
authors: KN
authors: Rajasekharan
authors: Bala
authors: B
authors: Nambisan
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D000935:Antifungal Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D001407:Bacillus / Q000378:metabolism*; D001419:Bacteria / Q000187:drug effects; D054659:Diketopiperazines / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D004151:Dipeptides / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D005285:Fermentation; D005658:Fungi / Q000187:drug effects*; D008826:Microbial Sensitivity Tests; D010456:Peptides, Cyclic / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D017172:Rhabditoidea / Q000382:microbiology*
chemical_list: D000900:Anti-Bacterial Agents; D000935:Antifungal Agents; D054659:Diketopiperazines; D004151:Dipeptides; D010456:Peptides, Cyclic