Medium Optimization for Penicillin Acylase (PAc) Production by Recombinant Containing pac Gene from Using Response Surface Methodology Bacillus megaterium MS941 Bacillus thuringiensis BGSC BD1 Using Response Surface Methodology  

Oleh:

Putri, Fentri Paramitha ; Nurhasanah, Astutiati ; Nurhayati, Niknik ; Helianti, Is ; Syamsu, Khaswar

Jenis: Article from Journal - ilmiah nasional - terakreditasi DIKTI
Dalam koleksi: Microbiology Indonesia vol. 9 no. 2 (Jun. 2015), page 65-72.
Topik: Bacillus megaterium btpacBD1; CCD; CaCl ; Penicillin acylase; xylose
Fulltext: 324-934_her.pdf (1.24MB)

Isi artikel

Penicillin G acylase (PAc) hydrolyses of the amide bond of benzylpenicillin (Pen-G) releasing PAA and 6APA, key intermediate in the production of various semisynthetic penicillins. In this study, we optimised the PAc production medium by RSM using two variables (xylose as inducer and CaCl as divalent cations) to obtain the optimum PAc specific activity from Bacillus megaterium btpacBD1. 2 the quadratic model was in good agreement with the experimental results (R = 0.86 (p-value < 0.0001)). 130.669 ± 50.241 units mg protein ) respectively, and medium pH was around 7. Under such conditions, the activity of PAc and protein concentration achieved were -1 1.318 ± 0.406 units mL and 0.01 ± 0.01 mg mL . The shake flask validation experiments demonstrated that with such medium composition the volumetric activity, protein concentration and specific activity achieved were -1 -1 1.294 ± 0.171 units mL , 0.0102 ± 0.0003 mg mL and 125.91 ± 13.31 units mg , respectively . When the optimum medium composition was applied in 10 L bioreactor, the optimum volumetric activity (2.0687 ± 0.0820 -1 -1 -1 units mL ) and protein concentration (0.0078 ± 0.0008 mg mL ) were achieved 48 h after the start of the -1 -1 cultivation. However, the optimum PAc specific acivity (1260.52 ± 27.57 units mg protein ) was achieved 18 h after the start of the cultivation. -1 For this purpose, combinations of five different xylose concentrations (0.13 – 0.87 %) and five different CaCl concentrations (0.64 – 4.36 mM) were 2 analysed, in a total of 22 experiments. CCD used for the analysis showed that in shake flask cultivations, xylose and CaCl gave significant effects on PAc volumetric activity and 2 The maximum specific activity ( was reached when xylose and CaCl concentrations were 0.49% and 2.4 mM,

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