玉米秸秆高效降解微生物复合菌系的构建及降解效果评价

doi:10.13560/j.cnki.biotech.bull.1985.2021-0968

摘要/Abstract

摘要：

针对玉米秸秆降解困难、利用率低的问题,本文筛选构建了由枯草芽孢杆菌WF-8（Bacillus subtilis WF-8）、地衣芽孢杆菌WF-11（B. lincheniformis WF-11）、蜡状芽孢杆菌WS-1（B. cereus WS-1）和黑胡桃链霉菌WF-10（Streptomyces nogalater WF-10）组成的高效降解复合菌系HD并进行秸秆降解效果评价,为高效降解玉米秸秆复合菌剂的研制和开发提供理论依据。本研究通过单因素试验结合正交试验和响应面试验,对复合菌系HD产酶培养基及发酵条件进行优化,液态和固态发酵试验评价复合菌系HD对玉米秸秆的降解效果。结果表明,复合菌系HD最佳碳氮源为秸秆粉和（NH₄）₂SO₄,最适添加量分别为15 g/L和4 g/L;复合菌系HD最优发酵条件为：温度34℃、接种量5%、时间6 d和pH7.2。优化后的复合菌系纤维素酶活为164.21 U/mL,是优化前的1.90倍,高于单菌酶活。将构建的复合菌系HD分别接种到液体发酵培养基和固体发酵培养基后,玉米秸秆降解率均显著高于单菌及对照组,降解率分别达到了47%和63.6%。研究结果表明复合菌系HD在降解玉米秸秆方面具有较大潜力,为高效降解玉米秸秆复合菌剂的研制和开发奠定了基础。

关键词: 复合菌系, 玉米秸秆降解, 纤维素酶活, 发酵条件优化

Abstract:

In response to the problems of difficult degradation and low utilization rate of corn straw,the efficient degradation microbial consortium HD composed of Bacillus subtilis WF-8,B. lincheniformis WF-11,B. cereus WS-1 and Streptomyces nogalater WF-10 was constructed,then the degradation effect was evaluated,which provided theoretical basis for the research and development of microbial compound agents of efficiently degrading corn stalk. Single factor experiment,orthogonal experiment and response surface method were combined to optimize the enzyme production medium and fermentation conditions of the microbial consortium HD. And experiments of liquid and solid fermentation were conducted to evaluate the degradation effect of the microbial consortium HD on corn straw. The results showed that the optimal carbon and nitrogen sources of microbial consortium HD were straw powder and（NH₄）₂SO₄,the optimal addition amounts were 15 and 4 g/L. And the optimal fermentation conditions were as follows：temperature 34℃,inoculating amount 5%,fermentation time 6 d and initial pH 7.2. Under this condition,the cellulase activity was 164.21 U/mL,1.9 times higher than that before optimization,which was significantly higher than that of a single strain. After inoculating the microbial consortium into liquid fermentation medium and solid fermentation medium,the degradation rate reached 47% and 63.6%,respectively;and the degradation rate was significantly higher than that of the single strain and control groups. The results indicate that the microbial consortium HD has great potential in the degradation of corn straw,which lays a foundation for the research and development of microbial consortium agents for the efficient degradation of corn straw.

Key words: microbial consortium, degradation of corn straw, activity of cellulase, optimization of fermentation conditions

王新光, 田磊, 王恩泽, 钟成, 田春杰. 玉米秸秆高效降解微生物复合菌系的构建及降解效果评价[J]. 生物技术通报, 2022, 38(4): 217-229.

WANG Xin-guang, TIAN Lei, WANG En-ze, ZHONG Cheng, TIAN Chun-jie. Construction of Microbial Consortium for Efficient Degradation of Corn Straw and Evaluation of Its Degradation Effect[J]. Biotechnology Bulletin, 2022, 38(4): 217-229.

图/表 14

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复合菌系 Microbial consortium	菌种名称 Name of strain
HA	WF-10、WF-11
HB	WF-10、WF-11、WF-8
HC	WF-10、WF-11、WS-1
HD	WF-10、WF-11、WF-8、WS-1

复合菌系 Microbial consortium	菌种名称 Name of strain
HA	WF-10、WF-11
HB	WF-10、WF-11、WF-8
HC	WF-10、WF-11、WS-1
HD	WF-10、WF-11、WF-8、WS-1

水平 Level	因素Factor
水平 Level	A碳源 Carbon source	B氮源 Nitrogen source	C碳源添加量 Content of carbon source/（g·L^-1）	D氮源添加量 Content of nitrogen source/（g·L^-1）
1	CMC-Na	酵母粉	5	1
2	纤维素	蛋白胨	10	3
3	秸秆粉	硫酸铵	12.5	4
4	淀粉	硝酸铵	15	5

水平 Level	因素Factor
水平 Level	A碳源 Carbon source	B氮源 Nitrogen source	C碳源添加量 Content of carbon source/（g·L^-1）	D氮源添加量 Content of nitrogen source/（g·L^-1）
1	CMC-Na	酵母粉	5	1
2	纤维素	蛋白胨	10	3
3	秸秆粉	硫酸铵	12.5	4
4	淀粉	硝酸铵	15	5

因素 Factor	水平 Level
因素 Factor	-1	0	+1
A温度/℃	30	35	40
B接种量/%	4	5	6
C时间/d	5	6	7
D初始pH	6	7	8