Sample Collections
The soil samples were collected from groundnut cultivated fields of Kadiri, Anantapur (Dist) at a depth of 10cm for soil sample.

Isolation of Soil Microorganism
The samples were immediately brought to the lab and air dried for 24 hours in a laminar air flow. For the isolation of bacteria, fungi, and actinomycetes, one gramme of dried soil was serially diluted and inoculated onto nutrient agar, potato dextrose agar, and starch casein agar at varied time intervals. Nutrient agar was incubated for 2 days, potato dextrose agar was incubated for 7 days, and starch casein agar was incubated for 5 days.

Preparation of Vanillin Production (VP) Medium¹¹
Composition of the Medium (g/L)
KH₂PO₄ – 0.9, Na₂ HPO₄ – 10, Mg SO₄.2H₂0 0.2, CaCl₂ – 0.01, FeSO₄. 2H₂0 – 0.1,MnSO₄ – 0.0006, Na₂MoSO₄ – 0.0006, Agar -15.0 and Bromothymol blue -0 .25, PH – 7.2, Ferulic acid was filter sterilised and put to sterile modified VP medium at a concentration of 0.1%. Pure strains were cultivated and incubated at 30°C for two days.

In Growing Cultures, Ferulic Acid is Bio Transformed into Vanillic Acid and Vanillin
The strains that developed on VP agar plates were selected and tested in VP broth at 30°C and 120 rpm for two days for bio vanillin and vanillic acid production spectrophotometer was used to quantify bacterial growth at 600 nm every 2 hours throughout the fermentation process, and a pH metre was used to monitor pH changes. Before being quantitatively analysed, the culture supernatant was collected and filtered using Whatmann filter paper.

Estimation of Bio Vanillin using TBA Reagent¹²
In an equivalent volume of 70 mM phosphate buffer, the cell free supernatant was diluted (500 L, pH 7) A solution of 24 percent HCl (5 mL) and 1 % thiobarbituric acid was used to acidify the mixture (2 mL) Heat for 10 minutes in a water bath at 55°C, then cool for 20 minutes at room temperature. A spectrophotometer was used to detect the absorbance at 434 nm, and a vanillin standard was used to estimate the equivalent vanillin content.¹³

Bioconversion of Ferulic Acid to Vanillin¹⁴
Luria Bertani (LB) agar was used to maintain pure cultures, which were incubated at 37°C for 24 to 48 hours. Bacillus subtilis colonies were then transferred to 100ml of LB broth (Luria Bertani) and cultured for 24-48 hours at 37°C. 1 % inoculum was added to the fermentation broth, which contained 15g of wheat bran and 0.05 % percent ferulic acid as a biotransformation inducer (g/100ml). (pH: 5.6.) Under shaking conditions, the fermentation broth was incubated at 37°C for 48-72 hours at 180 rpm. HPLC analysis was used to determine vanillin in the fermentation medium after 72 hours of biotransformation.

HPLC Analysis of Vanillin Produced in the Fermentation Medium¹⁵
Once biotransformation, the fermented material was centrifuged to eliminate the bran particles. 6N HCl was used to acidify the supernatant to a pH of 2. Three times with equal quantities of ethyl acetate, phenolic components were extracted. The supernatant was then used to determine the amount of vanillin using HPLC. The HPLC equipment was used to examine the generation of vanillin in the fermentation broth. The separation was carried out on a C18 column with a diameter of 4.6 x 250mm (5), a mobile phase of 90:10 acetonitrile in water, and a flow rate of 1.0ml/min. The material was filtered via a 0.22m membrane, and 20l of it was put straight into the HPLC column, where the products were identified at 230nm. For conventional vanillin, the separation was done at the same time.

Evaluation of Antimicrobial Activity of Vanillin Compound¹⁶
The antimicrobial activity of bacterial pathogens (acquired from SVIMS-Tirupati) vanillin compound was determined by well diffusion method. The compound of vanillin is placed in a well that contains 50µl, 100µl, Std (vanillin150µl) and control (Tetracycline).¹⁵ The plates were incubated for 24 hours at 37°C. The antimicrobial activity were done against Shigella, Bacillus, Staphylococcus, Enterococcus, and  Klebsiella.