Fifty fluorescent pseudomonads were isolated from rhizospheric soil of green gram from nearby area of Kaziranga, Assam, India and assayed for their extracellular proteinase production. Out of these isolates, 20 were found to be prominent in proteinase production. Genetic diversity of the 20 isolates were analyzed through BOX-PCR fingerprinting and 16S rDNA-RFLP along with three reference strains, viz., Pseudomonas fluorescens (NCIM2099(T)), Pseudomonas aureofaciens (NCIM2026(T)), and Pseudomonas aeruginosa (MTCC2582(T)). BOX-PCR produced two distinct clusters at 56% similarity coefficient and seven distinct BOX profiles. 16S rDNA-RFLP with three tetra-cutters restriction enzymes (HaeIII, AluI, and MspI) revealed two major clusters A and B; cluster A contained only single isolate FPS9 while the rest of 22 isolates belonged to the cluster B. Based on phenotypic characters and 16S rDNA sequence similarity, all the eight highly proteinase-producing strains were affiliated with P. aeruginosa. The proteinase was extracted from two most prominent strains (KFP1 and KFP2), purified by a three-step process involving (NH(4))(2)SO(4) precipitation, gel filtration, and ion exchange chromatography. The enzyme had an optimal pH of 8.0 and exhibit highest activity at 60°C and 37°C by KFP1 and KFP2 respectively. The specific activities were recorded as 75,050 (for KFP1) and 81,320 U/mg (for KFP2). The purified enzyme was migrated as a single band on native and SDS-PAGE with a molecular mass of 32 kDa. Zn(2+), Cu(2+), and Ni(2+) ion inhibited the enzyme activity. Enzyme activity was also inhibited by EDTA established as their metallo-proteinase nature.

• MeSH
• bakteriální proteiny genetika   metabolismus   MeSH
• endopeptidasy genetika   metabolismus   MeSH
• Fabaceae mikrobiologie   MeSH
• fylogeneze MeSH
• molekulární sekvence - údaje MeSH
• Pseudomonas klasifikace   enzymologie   genetika   izolace a purifikace   MeSH
• půdní mikrobiologie MeSH
• rhizosféra MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Indie MeSH

The most diverse and versatile endophytic actinobacteria are relatively unexplored potential sources of bioactive metabolites useful for different medical, agricultural, and other commercial applications. Their diversity in symbiotic association with traditionally utilized medicinal plants of northeast India is scantly available. The present investigation assessed the genetic diversity of endophytic actinobacteria (n = 120) distributed around the root, stem, and leaf tissues of six selected medicinal plants (Emblica officinalis, Terminalia chebula, T. arjuna, Murraya koenigii, Rauwolfia serpentina, and Azadirachta indica) from three different protected areas of evergreen forest-the Gibbon Wildlife Sanctuary (GWS), the Kaziranga National Park (KNP), and the North East Ecological Park (NEEP) of Assam, India. The samples were collected in two seasons (summer and winter). The overall phylogenetic analysis showed significant genetic diversity with 18 distinct genera belonging to 12 families. Overall, the occurrence of Streptomyces genus was predominant across all three sampling sites (76.66%), in both the sampling season (summer and winter). Shannon's and Simpson's diversity estimates showed their presence at A. indica (1.496, 0.778), R. serpentina (1.470, 0.858), and E. officinalis (0.975, 0.353). Among the site sampled, GWS had the most diverse community of actinobacteria (Shannon = 0.86 and Simpson = 0.557). The isolates were antagonistically more active against the investigated plant pathogenic bacteria than fungal pathogens. Further analysis revealed the prevalence of polyketide synthase genes (PKS) type II (84%) and PKS type I (16%) in the genome of the antimicrobial isolates. The overall findings confirmed the presence of biosynthetically active diverse actinobacterial members in the selected medicinal plants which offer potential opportunities towards the exploration of biologically active compounds.

• MeSH
• Actinobacteria klasifikace   genetika   izolace a purifikace   fyziologie   MeSH
• antibióza * MeSH
• Bacteria MeSH
• bakteriální proteiny genetika   metabolismus   MeSH
• endofyty klasifikace   genetika   izolace a purifikace   fyziologie   MeSH
• fylogeneze * MeSH
• fyziologie bakterií MeSH
• houby fyziologie   MeSH
• léčivé rostliny mikrobiologie   MeSH
• polyketidsynthasy genetika   metabolismus   MeSH
• roční období MeSH
• symbióza MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Indie MeSH