Biomolecular Characterization of Bacillus tequilensis A1C1 Isolated from Soil and Chromatographic Analysis of D-serine in its Cellular Fraction

The current work was carried out to investigate serine enantiomers in bacterial cells. The bacteria isolated from the pomace dumping soil site (bacteria id A1C1) showed maximum growth (O.D600 = 1.97±0.4 X 109cells/ml) within 48h in the minimal salt media supplemented with L-serine. The isolated strain was identified as ‘Bacillus tequilensis’ through 16sRNA sequencing. The study’s peculiarity reflects the fact that the isolated strain was explored for the first time to detect the presence of serine enantiomers. The strain was quantified for D-serine content by using RP-HPLC. The D-serine concentration was calculated as 0.919±0.02 nM in the bacterial cellular fraction by using a standard curve plot and linear curve equation. Further, recovery % was also calculated for the spiked samples which vary from 85-90%. The optimum growth parameters were recorded as 37℃±0.5, 150±0.5 RPM, and 7±0.5pH. The strain was Gram-positive, rod shape, large, irregular, off-white-coloured, and synthesized endospores. A1C1 showed positive results (within 14±2h of incubation) for indole production, lactose fermentation, and protease (0.9 mm, clear zone). The antibacterial assay showed 5% and 2% efficacy of the extracellular fraction against MTCC 40 and MTCC 11949 respectively within 12h of incubation. These results demonstrate that Bacillus tequilensis A1C1 has antibacterial activity, the potential to secrete extracellular enzymes, and D-serine content in the intracellular fraction of the cultivated cells. Given results demonstrate the industrial significance and implication of the isolated strain for the synthesis of commercially valuable products.

Bacillus tequilensis strain ‘UPMRB9’ improves biochemical attributes and nutrient accumulation in different rice varieties under salinity stress

Soil salinity exert negative impacts on agricultural production and regarded as a crucial issue in global wetland rice production (Oryza sativa L.). Indigenous salt-tolerant plant growth-promoting rhizobacteria (Bacillus sp.) could be used for improving rice productivity under salinity stress. This study screened potential salt-tolerant plant growth-promoting rhizobacteria (PGPR) collected from coastal salt-affected rice cultivation areas under laboratory and glasshouse conditions. Furthermore, the impacts of these PGPRs were tested on biochemical attributes and nutrient contents in various rice varieties under salt stress. The two most promising PGPR strains, i.e., ‘UPMRB9’ (Bacillus tequilensis 10b) and ‘UPMRE6’ (Bacillus aryabhattai B8W22) were selected for glasshouse trial. Results indicated that ‘UPMRB9’ improved osmoprotectant properties, i.e., proline and total soluble sugar (TSS), antioxidant enzymes like superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT). Moreover, ‘UPMRB9’ inoculated rice plants accumulated higher amount of nitrogen and calcium in tissues. Therefore, the indigenous salt-tolerant PGPR strain ‘UPMRB9’ could be used as a potential bio-augmentor for improving biochemical attributes and nutrient uptake in rice plants under salinity stress. This study could serve as a preliminary basis for future large-scale trials under glasshouse and field conditions.

Biomolecular Characterization of Bacillus tequilensis A1C1 Isolated from Soil and Chromatographic Analysis of D-serine in its Cellular Fraction

The bacteria isolated from the pomace dumping soil site (bacteria id A1C1) showed maximum growth (O.D600 = 1.97±0.4 X 109 cells/ml) within 48h in the minimal salt media supplemented with L-serine. The isolated strain was identified as ‘Bacillus tequilensis’ through 16sRNA sequencing. The strain was quantified for D-serine content by using RP-HPLC. The D-serine concentration was calculated as 0.919±0.02 nM in the bacterial cellular fraction by using a standard curve plot and linear curve equation. Further, recovery % was also calculated for the spiked samples which vary from 85-90%. The study’s peculiarity reflects the fact that the isolated strain was explored for the first time to detect the presence of serine enantiomers. The biochemical features also showed 70% similarity to the standard strain Bacillus tequilensis 10bT. The optimum growth parameters were recorded as 37℃±0.5, 150±0.5 RPM, and 7±0.5pH. The strain was Gram-positive and synthesized endospores. Morphological results showed its rod shape, large, irregular, and off-white-coloured colonies. A1C1 was also tested for the production of secondary metabolites and enzymes. A1C1 showed positive results for indole production, lactose fermentation, protease, and gelatinase whereas, negative results for catalase, MR-VP, citrate utilization, cellulase, amylase, and pectinase. Further, the strain was assayed for PGPR attributes and showed a negative phosphate solubilization index and IAA production. The antibacterial assay showed 5% and 2% efficacy of the extracellular fraction against MTCC 40 and MTCC 11949 respectively. These results demonstrate that Bacillus tequilensis A1C1 has antibacterial activity, the potential to secrete extracellular enzymes, and D-serine content in the intracellular fraction of the cultivated cells.

Biomolecular Characterization of Bacillus tequilensis A1C1 Isolated from Soil and Chromatographic Analysis of D-serine in its Cellular Fraction

The bacteria isolated from the pomace dumping soil site (bacteria id A1C1) showed maximum growth (O.D600 = 1.97±0.4 X 109 cells/ml) within 48h in the minimal salt media supplemented with L-serine. The isolated strain was identified as ‘Bacillus tequilensis’ through 16sRNA sequencing. The strain was quantified for D-serine content by using RP-HPLC. The D-serine concentration was calculated as 0.919±0.02 nM in the bacterial cellular fraction by using a standard curve plot and linear curve equation. Further, recovery % was also calculated for the spiked samples which vary from 85-90%. The study’s peculiarity reflects the fact that the isolated strain was explored for the first time to detect the presence of serine enantiomers. The biochemical features also showed 70% similarity to the standard strain Bacillus tequilensis 10bT. The optimum growth parameters were recorded as 37℃±0.5, 150±0.5 RPM, and 7±0.5pH. The strain was Gram-positive and synthesized endospores. Morphological results showed its rod shape, large, irregular, and off-white-coloured colonies. A1C1 was also tested for the production of secondary metabolites and enzymes. A1C1 showed positive results for indole production, lactose fermentation, protease, and gelatinase whereas, negative results for catalase, MR-VP, citrate utilization, cellulase, amylase, and pectinase. Further, the strain was assayed for PGPR attributes and showed a negative phosphate solubilization index and IAA production. The antibacterial assay showed 5% and 2% efficacy of the extracellular fraction against MTCC 40 and MTCC 11949 respectively. These results demonstrate that Bacillus tequilensis A1C1 has antibacterial activity, the potential to secrete extracellular enzymes, and D-serine content in the intracellular fraction of the cultivated cells.

Biomolecular Characterization of Bacillus tequilensis A1C1 Isolated from Soil and Chromatographic Analysis of D-serine in its Cellular Fraction

The bacteria isolated from the pomace dumping soil site (bacteria id A1C1) showed maximum growth (O.D600 = 1.97 ± 0.4 X 109 cells/ml) within 48h in the minimal salt media supplemented with L-serine. The isolated strain was identified as ‘Bacillus tequilensis’ through 16sRNA sequencing. The strain was quantified for D-serine content by using RP-HPLC. The D-serine concentration was calculated as 0.919 ± 0.02 nM in the bacterial cellular fraction by using a standard curve plot and linear curve equation. Further, recovery % was also calculated for the spiked samples which vary from 85–90%. The study’s peculiarity reflects the fact that the isolated strain was explored for the first time to detect the presence of serine enantiomers. The biochemical features also showed 70% similarity to the standard strain Bacillus tequilensis 10bT. The optimum growth parameters were recorded as 37℃±0.5, 150 ± 0.5 RPM, and 7 ± 0.5pH. The strain was Gram-positive and synthesized endospores. Morphological results showed its rod shape, large, irregular, and off-white-coloured colonies. A1C1 was also tested for the production of secondary metabolites and enzymes. A1C1 showed positive results for indole production, lactose fermentation, protease, and gelatinase whereas, negative results for catalase, MR-VP, citrate utilization, cellulase, amylase, and pectinase. Further, the strain was assayed for PGPR attributes and showed a negative phosphate solubilization index and IAA production. The antibacterial assay showed 5% and 2% efficacy of the extracellular fraction against MTCC 40 and MTCC 11949 respectively. These results demonstrate that Bacillus tequilensis A1C1 has antibacterial activity, the potential to secrete extracellular enzymes, and D-serine content in the intracellular fraction of the cultivated cells.

Optimasi Kondisi Fermentasi pada Produksi Metabolit Antibakteri dari Bacillus tequilensis BSMF Simbiotik Halichondria panicea

Pendahuluan: Resistensi antibakteri merupakan masalah kesehatan global yang dialami hampir di seluruh negara. Eksplorasi antibakteri dari sumber baru seperti tumbuhan, hewan, dan mikroorganisme baik yang hidup bebas maupun bersimbiosis menjadi solusi alternatif untuk mengatasi resistensi antibakteri. Mikroorganisme berupa bakteri ditargetkan sebagai sumber antibakteri yang berkelanjutan karena jumlahnya melimpah dan mudah dalam proses pembiakan. Bakteri yang hidup bersimbiosis diketahui dapat memproduksi metabolit antibakteri berspektrum lebih luas dibandingkan bakteri yang hidup bebas. Bakteri dapat bersimbiosis dengan berbagai makhluk hidup termasuk organisme multiseluler seperti spons. Isolat Bacillus tequilensis BSMF yang bersimbiosis dengan Halichondria panicea dari Perairan Cabbiya Madura menunjukkan adanya produksi metabolit yang memiliki aktivitas antibakteri. Tujuan: Menentukan pH dan suhu optimum untuk produksi metabolit antibakteri dari Bacillus tequilensis BSMF simbiotik Halichondria panicea. Metode: Produksi metabolit antibakteri dilakukan dengan metode fermentasi padat pada media Potato Dextrose Agar (PDA) yang telah diatur pH dan suhu inkubasinya, sedangkan uji aktivitas antibakteri terhadap Staphylococcus aureus ATCC 25923 dan Eschericia coli ATCC 25922 dilakukan menggunakan metode difusi agar. Penentuan aktivitas antibakteri dilakukan melalui pengukuran diameter zona hambat. Hasil: pH media yang menunjukkan aktivitas antibakteri optimum Bacillus tequilensis BSMF terhadap Staphylococcus aureus ATCC 25923 dan Eschericia coli ATCC 25922 adalah 8 ± 0,5 pada suhu inkubasi 32 ± 1oC dengan rata- rata indeks aktivitas antibakteri berturut- turut 2,74 ± 0,07 dan 3,39 ± 0,07. Kesimpulan: pH dan suhu optimum yang diperoleh adalah pH 8 ± 0,5 dan suhu 32 ± 1oC.