响应面法优化黑曲霉HDF05产β-葡萄糖苷酶过程参数
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国家高技术研究发展计划 (863计划) (No. 2007AA100702-6),黑龙江省科技攻关重大项目 (No. GA07B401-6),哈尔滨市科技创新人才研究专项资金 (No. RC2010XK002028),黑龙江省教育厅重点项目 (No. 11551z011),国家自然科学基金 (No. 31070446),黑龙江大学高层次人才 (创新团队) 支持计划 (No. hdtd2010-17) 资助。


Fermentation optimization by response surface methodology for enhanced production of β-glucosidase of Aspergillus niger HDF05
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National High Technology Research and Development Program of China (863 Program) (No. 2007AA100702-6), Key Technology Research and Development Program of Heilongjiang Science and Technology Bureau (No. GA07B401-6), Special Fund for Scientific and Technolo

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    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%.

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凌宏志,葛菁萍,平文祥,许修宏. 响应面法优化黑曲霉HDF05产β-葡萄糖苷酶过程参数[J]. 生物工程学报, 2011, 27(3): 419-426

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  • 收稿日期:2010-08-02
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