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Molecular characterization of a proteolytic bacterium in Panchagavya: An organic fertilizer mixture.

ABSTRACT: Fermented product of combination of five major substances obtained from cow, viz., urine, milk, ghee, curd, and dung, is known as Panchagavya. Its pro-agricultural and medicinal value has been traditionally known to Indian farmers from Vedic period. In this study, the proteolytic properties of Panchagavya were investigated using Skim Milk Agar (SMA) form, a commercially available Panchagavya product. Proteolytic bacteria, SNCK-3, was successfully isolated. Further identification using 16s rDNA sequencing revealed that SNCK-3 belonged to Acinetobacter spp., which is a species of biofertilizer group. This observation justified the pro-agricultural role of Panchagavya. The present study represents primary data and it is essential to develop a new area of research for exposing the invisible or dormant Vedic biotechnological concepts, like Panchagavya.

SUBMITTER: Sayi DS

PROVIDER: S-EPMC6034162 | biostudies-literature | 2018 Apr - Jun

REPOSITORIES: biostudies-literature

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Molecular characterization of a proteolytic bacterium in Panchagavya: An organic fertilizer mixture.

Sayi D S DS Mohan Surya S Vinod Kumar K K

Journal of Ayurveda and integrative medicine 20171206 2

Fermented product of combination of five major substances obtained from cow, viz., urine, milk, ghee, curd, and dung, is known as Panchagavya. Its pro-agricultural and medicinal value has been traditionally known to Indian farmers from Vedic period. In this study, the proteolytic properties of Panchagavya were investigated using Skim Milk Agar (SMA) form, a commercially available Panchagavya product. Proteolytic bacteria, SNCK-3, was successfully isolated. Further identification using 16s rDNA s ...[more]

PMID: 29203350

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Project description:Flagellins are the protein components of bacterial flagella and assemble in up to 20,000 copies to form extracellular flagellar filaments. An unusual family of flagellins was recently discovered that contains a unique metalloprotease domain within its surface-exposed hypervariable region. To date, these proteolytic flagellins (also termed flagellinolysins) have only been characterized in the Gram-positive organism Clostridium haemolyticum, where flagellinolysin was shown to be proteolytically active and capable of cleaving extracellular protein substrates. The biological function of flagellinolysin and its activity in other organisms, however, remain unclear. Here, using molecular biochemistry and proteomics, we have performed an initial characterization of a novel flagellinolysin identified from Hylemonella gracilis, a Gram-negative organism originally isolated from pond water. We demonstrate that H. gracilis flagellinolysin (HgrFlaMP) is an active calcium-dependent zinc metallopeptidase and characterize its cleavage specificity profile using both trypsin and GluC-derived peptide libraries and protein substrates. Based on high-throughput degradomic assays, HgrFlaMP cleaved 784 unique peptides and displayed a cleavage site specificity similar to flagellinolysin from C. haemolyticum. Additionally, by using a set of six protein substrates, we identified 206 protein-embedded cleavage sites, further refining the substrate preference of HgrFlaMP, which is dominated by large hydrophobic amino acids in P1', and small hydrophobic or medium-sized polar residues on the amino-terminal side of the scissile bond. Intriguingly, recombinant HgrFlaMP was also capable of cleaving full-length flagellins from another species, suggesting its potential involvement in interbacterial interactions. Our study reports the first experimentally characterized proteolytic flagellin in a Gram-negative organism, and provides new insights into flagellum-mediated enzymatic activity.

Project description:Decomposed organic materials, in combination with plant growth-promoting bacteria (PGPB), are environmentally friendly and reduce synthetic fertilizer use in rice production. A bio-organic fertilizer (BoF) was prepared using kitchen waste (79%), chita-dhan (unfilled rice grain) biochar (15%), rock phosphate (5%), and a consortium of 10 PGPB (1%) to supplement 30% nitrogen and to replace triple superphosphate (TSP) fertilizer in rice production with an improvement of soil health. PGPB were local isolates and identified using 16S ribosomal RNA partial gene sequences as Bacillus mycoides, Proteus sp., Bacillus cereus, Bacillus subtilis, Bacillus pumilus, Paenibacillus polymyxa, and Paenibacillus spp. Isolates could fix N2 by 0.7-1.4 g kg-1, solubilize 0.1-1.2 g kg-1 phosphate, and produce 0.1-40 g kg-1 indoleacetic acid. The performance of BoF was evaluated by 16 field experiments and 18 farmers' field demonstration trials during the year 2017-2020 in different parts of Bangladesh. Performances of BoF were evaluated based on control (T1), full synthetic fertilizer dose of N, P, and K (T2), BoF (2 t ha-1) + 70% N as urea + 100% K as muriate of potash (T3), 70% N as urea + 100% P as TSP + 100% K as muriate of potash (T4), and 2 t ha-1 BoF (T5) treatments. At the research station, average grain yield improved by 10-13% in T3 compared with T2 treatment. Depending on seasons, higher agronomic N use efficiency (19-30%), physiological N use efficiency (8-18%), partial factor productivity (PFP)N (114-150%), recovery efficiency (RE)N (3-31%), N harvest index (HIN) (14-24%), agronomic P use efficiency (22-25%), partial factor productivity of P (9-12%), AREP (15-23%), and HIP (3-6%) were obtained in T3 compared with T2 treatment. Research results were reflected in farmers' field, and significant (P < 0.05) higher plant height, tiller, panicle, grain yield, partial factor productivity of N and P were obtained in the same treatment. Application of BoF improved soil organic carbon by 6-13%, along with an increased number of PGPB as compared with full synthetic fertilizer dose. In conclusion, tested BoF can be considered as a green technology to reduce 30% synthetic N and 100% TSP requirements in rice production with improved soil health.

Dataset Information

Molecular characterization of a proteolytic bacterium in Panchagavya: An organic fertilizer mixture.

Publications

Molecular characterization of a proteolytic bacterium in Panchagavya: An organic fertilizer mixture.

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