Laboratory of Immunopathology Keizo Asami (LIKA), Federal University of Pernambuco, Av. Prof. Moraes Lins do Rego, s/n, 50670-901, Recife, PE, Brazil.
Biotechnol Prog. 2011 Sep-Oct;27(5):1470-7. doi: 10.1002/btpr.664. Epub 2011 Jul 19.
Medium composition and culture conditions for maximal collagenase production by Penicillium aurantiogriseum URM4622 were optimized using a response surface approach. A full two-level design on three factors (initial medium pH, soybean flour concentration, and temperature) was employed to identify the most significant fermentation parameters for collagenase production, and a subsequent central composite design (CCD) was used to find the optimal levels of the two most significant factors (initial medium pH and soybean flour concentration). The design results indicated that the initial medium pH and the temperature had significant negative main effects, whereas the substrate concentration had a positive effect on the collagenase production. The maximum collagenolytic activity predicted by the fitted response surface was expected to occur at pH 7.21, 1.645% soybean flour concentration and 24°C. Three replicate experiments were run at these conditions and yielded an activity response of 283.36 ± 1.33 U, which not only is the highest obtained in this study but also represents a 5-fold increase over the lowest response observed in the initial design. Since all experiments were carried out with an inexpensive substrate, the final results point out to a cost-effective medium for collagenase production with potential industrial-scale applications.
采用响应面法优化了橙色青霉 URM4622 产生胶原酶的培养基组成和培养条件。采用三因素（初始培养基 pH 值、豆粉浓度和温度）的完全两水平设计来确定对胶原酶生产最显著的发酵参数，并随后采用中心复合设计（CCD）来确定两个最显著因素（初始培养基 pH 值和豆粉浓度）的最佳水平。设计结果表明，初始培养基 pH 值和温度有显著的负主效应，而基质浓度对胶原酶的产生有积极的影响。拟合响应面预测的最大胶原酶活力预计将在 pH7.21、1.645%豆粉浓度和 24°C 时出现。在这些条件下进行了三次重复实验，得到的活性响应为 283.36±1.33U，不仅是本研究中获得的最高值，而且比初始设计中观察到的最低值提高了 5 倍。由于所有实验都是用廉价的基质进行的，最终结果表明，这种培养基具有成本效益，可用于胶原酶的生产，具有潜在的工业应用前景。
