**2. Material and methods**

#### **2.1 Chemicals and reagents**

BPA, hydrochloric acid, sodium acetate, sodium hydroxide, calcium chloride, iron sulphate, potassium phosphate, hydrogen peroxide, ammonium nitrate, and 2,2′azino-di-(3 ~ ethyl benzothiazoline-6-sulphonic acid) (ABTS) were products of Sigma-Aldrich, Germany. Ethanol and yeast extract were ordered from BDH Chemicals Limited, England, Scharlaun Chemie S.A. Barcelona, Spain, and Biomark Laboratories Pune, India. All the other chemicals and reagents used were of analytical grade.

#### **2.2 Microorganism**

Sixteen actinobacterial isolates that were obtained from the culture collection of the Enzyme and Microbial Technology laboratory, Department of Biochemistry, Federal University of Technology, Akure, Nigeria, were screening for this study. Three (*A. naeslundii, A. bovis*, and *A. israelii*) of the isolates with best-growing ability on media supplemented with 100 mg/L of BPA were selected from the culture bank for further studies.

#### *2.2.1 Seed culture preparation*

Seed culture for each actinobacterial strain was prepared by growing a loopful portion from the slant in sterile media containing (5 g/L), yeast extract (1.5 g/L),

#### *A Laboratory-Scale Study: Biodegradation of Bisphenol A (BPA) by Different Actinobacterial… DOI: http://dx.doi.org/10.5772/intechopen.105546*

beef extract (1.5 g/L), and NaCl (5 g/L) with pH adjusted to 7.4. The media were incubated in an orbital incubator at 30°C for 24 h at 160 r⋅min−1 in a shaking incubator (Stuart, UK). After 24 h, the seed culture was used as inoculum for the production media. Each seed inoculum (constituting 5% v/v) was transferred into 500-mL Erlenmeyer flask containing 100 mL of production media. Three different actinobacterial consortia were developed as follows: *A. naeslundii, A. israelii*, and *A. bovis* used as inoculant for the degradation processes.
