TY - JOUR AU - PUTRI, FENTRI PARAMITHA AU - NURHASANAH, ASTUTIATI AU - NURHAYATI, NIKNIK AU - HELIANTI, IS AU - SYAMSU, KHASWAR PY - 2015/06/30 Y2 - 2024/03/29 TI - Medium Optimization for Penicillin Acylase (PAc) Production by Recombinant B. megaterium MS941 Containing pac Gene from B. thuringiensis BGSC BD1 Using Response Surface Methodology JF - Microbiology Indonesia JA - Microbiol indones VL - 9 IS - 2 SE - Articles DO - 10.5454/mi.9.2.3 UR - https://jurnal.permi.or.id/index.php/mionline/article/view/324 SP - 3 AB - <p>Penicillin G acylase (PAc) hydrolyses of the amide bond of benzylpenicillin (Pen-G) releasing PAA and 6-APA, key intermediate in the production of various semisynthetic penicillins. In this study, we optimised the production medium of PAc by RSM using two variables (xylose as inducer and CaCl<sub>2</sub> as divalent cations) to obtain the optimum PAc specific activity from <em>Bacillus megaterium</em> bt<em>pac</em>BD1. For this purpose, combinations of five different xylose concentrations (0.13 – 0.87 %) and five different CaCl<sub>2</sub> concentrations (0.64 – 4.36 mM) were analysed, in a total of 22 experiments. CCD used for the analysis showed that in shake flask cultivations, xylose and CaCl<sub>2</sub> showed significant effects on PAc volumetric activity and the quadratic model was in good agreement with the experimental results (R<sup>2</sup>= 0.86 (p-value &lt; 0.0001)). The maximum specific activity (130.669 ± 50.241 units mg protein<sup>-1</sup>) was reached when xylose and CaCl<sub>2</sub> concentrations were 0.49% and 2.4 mM, respectively, and medium pH was around 7. Under such conditions, the activity of PAc and protein concentration achieved were 1.318 ± 0.406 units mL<sup>-1</sup> and  0.0101 ± 0.01 mg mL<sup>-1</sup>. The shake flask validation experiments demonstrated that with such medium composition the volumetric activity, protein concentration and specific activity achieved were 1.294 ± 0.171 units mL<sup>-1</sup>, 0.0102 ± 0.0003 mg mL<sup>-1</sup> and 125.91 ± 13.309 units mg<sup>-1</sup>, respectively. When the optimum medium composition was applied in 10 L bioreactor, the optimum volumetric activity (2.0687 ± 0.0820 units mL<sup>-1</sup>) and protein concentration (0.0078 ± 0.0008 mg mL<sup>-1</sup>) were achieved 48 h after the start of the cultivation. However, the optimum PAc specific acivity (1260.52  ± 27.5711 units mg protein<sup>-1</sup>) was achieved 18 h after the start of the cultivation.</p> ER -