1.

Experimental details

Biosynthesis of AgNP´s with Melissa officinalis extract

For the biosynthesis of Ag NPs, Silver nitrate AgNO 3 (99.98 %) reagent grade was used as precursor (Sigma-

Aldrich). The Melissa officinalis employed as reducing agent was obtained from Morelia City, México. Briefly,

0.01274 gr of AgNO3 were dissolved with 15 ml (5 mM) of deionized water. For the Melissa officinalis extract, the

leaves were dried by exposure to sun. Subsequently, 5 g of the dried leaves was milled and transferred to a 50 ml

flask with 20 ml of distilled water. The Melissa officinalis solution was heated to 100 °C for 10 min. The infusion

obtained was mixed whit 10 ml of AgNO3 (5 mM) solution at room temperature. After 1 hour, a color change of the

post-reaction mixtures from light green into dark brown was observed.

Structural Characterization.

The samples were characterized by scanning electron microscopy (SEM) using the Hitachi SU8230 cold field

emission (CFE) SEM/STEM microscope. The structural characterization of the AgNP`s was carried out using an X-

ray diffractometer (Rigaku Ultima IV, using Cu-K radiation). Analysis of Fourier transform infrared (FT-IR) and

Raman (RMN) spectroscopy were measured at room temperature using a Bruker Spectrometer (RFS 100/S). The

Uv-Vis spectroscopy technique was employed to support the AgNP`s formation by a fiber optic spectrometer

(Ocean Optics model USB4F01962).

Bioprocess Biosyst Eng

Antibacterial assay.

The antibacterial activity of the silver nanoparticles were assessed against

Escherichia Coli (E. coli) and Staphylococcus Aureus (S. Aureus). The antibacterial

test was carried out via a growth inhibition assay. Six filter paper discs of 5 mm of

diameter were impregnated with 5 ml de AgNP´s solutions at concentrations of 1, 5,

9, 10, 15 and 20 mM. Subsequently, the discs were placed in the vial containing the

E. coli And S. Aureus. The system was incubated at 36 °C for 24 h. Finally, the

inhibition zone was measured.

Frecuency (%)
Counts (a.u.)

600 10

0
0 4 8 12 16 20 24 28
Particle Size (nm)
Ag
300