响应面法优化黑曲霉HDF05产β-葡萄糖苷酶过程参数

Fermentation optimization by response surface methodology for enhanced production of β-glucosidase of Aspergillus niger HDF05

摘要:

为获得黑曲霉Aspergillus niger HDF05菌株较高的β-葡萄糖苷酶酶活，对其发酵条件进行了优化。采用Plackett-Burman实验设计考察关键发酵操作参数对产酶的影响。继而采用最陡爬坡路径逼近最大响应区域，并结合中心组合实验和响应面对4个显著性因素进行分析。Plackett-Burman实验结果表明，发酵温度、装液量、麦麸和 (NH4)2SO4浓度对β-葡萄糖苷酶合成影响显著。通过响应面分析得到一元二阶方程，对方程求解得到优化的发酵过程参数：发酵温度为28 ℃，装液量为71.4 mL/250 mL，麸皮浓度为36 g/L，(NH4)2SO4浓度为5.5 g/L。采用该优化的过程参数，菌株的最大产β-葡萄糖苷酶活力可达60.06 U/mL，较优化前提高了23.9％。将黑曲霉HDF05产生的β-葡萄糖苷酶用于酸解玉米芯纤维残渣的酶解实验中，可明显降低纤维二糖的积累，48 h内可使玉米芯纤维素残渣酶解得率达到80.4％。

Abstract:

In order to obtain high β-glucosidase productivity, we optimized the fermentation parameters for β-glucosidase production by Aspergillus niger HDF05. First, we screened the important parameters by Plackeet-burman design. Second, we used the path of steepest ascent to approach to the biggest response region of parameters affecting β-glucosidase production. Then, we obtained the optimal parameters by central composite design and response surface analysis. We developed a quadratic polynomial equation for predicting β-glucosidase production level. The results showed that the important parameters were temperature, packing volume, concentrations of wheat bran and (NH4)2SO4. The optimal fermentation parameters were as follows: temperature 28 °C, packing volume 71.4 mL/250 mL, wheat bran 36 g/L and (NH4)2SO4 5.5 g/L. Under the optimal conditions, we obtained the maximum enzyme activity of 60.06 U/mL, with an increase of 23.9% compared to the original fermentation parameters. During enzymatic hydrolysis of acid-pretreated corncob, addition of β-glucosidase from Aspergillus niger HDF05 greatly reduced the inhibition caused by cellobiose, and the hydrolysis yield was improved from 66.7% to 80.4%.