Xinjiang Agricultural Sciences ›› 2018, Vol. 55 ›› Issue (11): 2122-2133.DOI: 10.6048/j.issn.1001-4330.2018.11.020

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Study on Optimizing Liquid Fermentation Conditions of Aspergillus niger and Its Degradation Effect on Cotton Stalk with Double Indexes

GUO Kai1,2, HOU Min2, BAO Hui-fang2, WANG Ning2, ZHAN Fa-qiang2, YANG Rong2, YANG Wen-qi2, LONG Xuan-qi2, CUI Wei-dong2   

  1. 1.College of Life Science and Technology, Xinjiang University, Urumqi 830046, China;
    2. Research Institute of Applied Microbiology, Xinjiang Academy of Agricultural Sciences / Xinjiang Laboratory of Special Environmental Microbiology, Urumqi 830091, China
  • Received:2018-09-30 Published:2019-02-22
  • Correspondence author: CUI Wei-dong (1969-),researcher,master's degree,Research direction fermentation engineering. E-mail:850530399@qq.com
  • Supported by:
    Youth Fund Project of Xinjiang Uygur Autonomous Region"Study on the Degradation Effect of Lignocellulose Degrading Strains and Mixed Fermentation in Cotton Stalk Feed"(2015211B019); Basic Scientific R & D Program of Public Welfare Research Institutes of Xinjiang Uygur Autonomous Region of China "Study on Microbial Treatment Technology of Cotton Straw Feeding"(KY2014028)

双指标优化黑曲霉液态发酵条件及降解棉秆效果

郭凯1,2, 侯敏2, 包慧芳2, 王宁2, 詹发强2, 杨蓉2, 杨文绮2, 龙宣杞2, 崔卫东2   

  1. 1.新疆大学生命科学与技术学院,乌鲁木齐 830052;
    2.新疆农业科学院微生物应用研究所/新疆特殊环境微生物实验室,乌鲁木齐 830091
  • 通讯作者: 崔卫东(1969-),男,研究员,硕士,研究方向为发酵工程,E-mail:850530399@qq.com
  • 作者简介:郭凯(1993-),男,硕士研究生,研究方向为微生物发酵工程,E-mail:guokairn@sohu.com
  • 基金资助:
    新疆维吾尔自治区青年基金项目“木质纤维素降解菌株选育及混菌发酵在棉秆饲料中降解效果研究”(2015211B019);自治区公益性科研院所基本科研业务经费资助项目“棉秸秆饲料化微生物处理技术研究”(KY2014028)

Abstract: 【Objective】 This project aims to investigate the degradation effect of Aspergillus niger on cotton straw by using the Carboxymethylcellulose sodium enzyme activity,(CMCase) and the Filter paper enzyme activity (FPAase) as two indexes to optimize the liquid fermentation conditions of Aspergillus niger ZD to produce cellulase by response surface methodology.【Method】The main influencing factors were initially determined by single factor experiment, and through the optimization of the factors and interaction by Plackett-Burman and Box-Behnken, the cellulose content in cotton straw was determined. 【Result】The single factor experiment determined that the carbon source was starch, the nitrogen source was peptone, the inorganic salt was K2HPO4, the culture time was 168 h, the rotation speed was 150 r/min, the temperature was 30℃, and the inoculum volume was 7%. The significant factors affecting enzyme production selected by Plackett-Burman design were: starch mass concentration, peptone mass concentration, rotational speed and inoculation amount. Finally, the optimal fermentation conditions for enzyme production were determined by Box-behnken, the starch mass concentration was 1.21 g/100 mL, the peptone mass concentration was 0.25 g/100 mL, and the K2HPO4 mass concentration was 0.1 g. /100 mL, culture time 168 h, speed 170 r/min, temperature 30℃, inoculum 7.8%, 35.96% higher than the initial fermentation conditions. 【Conclusion】The use of Aspergillus niger to degrade cotton stalks by liquid fermentation is effective, because it can effectively degrade cellulose and hemicellulose in cotton stalks, which shows that it is promising to use fungi to make stalks to forage.

Key words: Aspergillus niger; liquid fermentation; cellulase; cotton stalk; hemicellulose

摘要: 【目的】以羧甲基纤维素钠酶活(Carboxymethylcellulose sodium enzyme activity,CMCase)和滤纸酶活(Filter paper enzyme activity,FPAase)作为双指标,通过响应面法对黑曲霉(Aspergillus niger)ZD产纤维素酶的液体发酵条件进行优化,研究黑曲霉对棉花秸秆的降解效果。【方法】通过单因素试验确定主要影响因素,通过Plackett-Burman和Box-Behnken设计进行因素优化和交互作用的影响,测定棉秆中纤维素含量。【结果】单因素试验确定碳源为淀粉,氮源为蛋白胨,无机盐为K2HPO4,培养时间168 h,转速150 r/min,温度30℃,接种量7%;利用Plackett-Burman设计筛选出影响产酶的显著因素是淀粉质量浓度、蛋白胨质量浓度、转速和接种量;最后通过Box-behnken确定产酶的最佳发酵条件,淀粉质量浓度1.21 g/100 mL,蛋白胨质量浓度0.25 g/100 mL,K2HPO4质量浓度0.1 g/100 mL,培养时间168 h,转速170 r/min,温度30℃,接种量7.8%,比初始发酵条件提高了35.96%。【结论】利用黑曲霉通过液体发酵降解棉秆效果显著,可以有效降解棉秆中纤维素和半纤维素,用真菌实现棉秆饲草化极具前景。

关键词: 黑曲霉, 液体发酵, 纤维素酶, 棉花秸秆, 半纤维素

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