Screening of strains producing non-starch polysaccharide enzyme by fermentation of licorice residue and its dynamic changes

doi:10.6048/j.issn.1001-4330.2024.11.014

Abstract

Abstract:

【Objective】 To screen a strain with high yield of non-starch polysaccharide enzyme four fungi in the hope of laying a foundation for the production of non-starch polysaccharide enzyme by solid-state fermentation of licorice residue.【Methods】 Four kinds of fungi (Aspergillus niger, Trichoderma reesei, Phanerochaete chrysosporium and Trichoderma koningii) were primarily screened by Congo red method and filter paper strip disintegration test, and the CMC enzyme, filter paper enzyme and xylanase activities were determined by shake flask fermentation of residue.The strain with the highest enzyme activity was fermented with licorice residue in solid state to verify its enzyme production effect.【Results】 Through the preliminary screening, it was found that all four strains of fungi had the ability to produce cellulase and xylanase.The antagonistic effects between Aspergillus Niger and other three strains was found by plate confrontation test.According to the results of enzyme activity determination, Phanerochaete chrysosporium and Trichoderma koningii produced the highest cellulase than the other two fungi, and the CMC enzyme activity reached 2.10 and 2.31 IU/mL on the fourth day of fermentation.The highest xylanase activity produced by Aspergillus Niger through shake flask re-screening fermentation could reach 97.46 IU/mL. However, the xylanase activity of Phanerochaete chrysosporium and Trichoderma koningii could reach 131.707 IU/mL at the highest.【Conclusion】 The growth cycle of non-starch polysaccharide activity produced by mixed solid-state fermentation of licorice residue with Phanerochaete chrysosporium and Trichoderma koningii is more stable and the yield is higher, therefore, licorice residue, as an agricultural by-product, has the potential to become a low-cost substrate for enzyme production in the future.

Key words: licorice residue; cellulase; xylanase; solid state fermentation

摘要：

【目的】从4株真菌中筛选出产非淀粉多糖酶水平较强的菌株,为甘草渣固态发酵生产非淀粉多糖酶工艺奠定基础。【方法】设计刚果红法及滤纸条崩解试验,对4种真菌(黑曲霉、里氏木霉、黄孢原毛平革菌、康宁木霉)进行初筛,采用甘草渣摇瓶发酵进行复筛,测定纤维素酶(CMC酶、滤纸酶)及木聚糖酶活力。将产酶水平最高的菌株与甘草渣进行固态发酵,验证其产酶效果。【结果】4株真菌均具有产纤维素酶、木聚糖酶的能力。黑曲霉与其他3种菌株间存在拮抗作用。黄孢原毛平革菌和康宁木霉较其他2株真菌产纤维素酶较高,在发酵第4 d时CMC酶活力分别达到2.10、2.31 IU/mL。黑曲霉摇瓶复筛发酵产木聚糖酶活力最高可达97.46 IU/mL,而黄孢原毛平革菌与康宁木霉混菌发酵产木聚糖酶活力最高可达131.707 IU/mL。【结论】甘草渣与黄孢原毛平革菌、康宁木霉混合固态发酵生产出的非淀粉多糖酶活力增长周期更稳定,产量更高。甘草渣作为农业副产品有潜质成为未来生产酶的低成本底物。

关键词: 甘草渣, 纤维素酶, 木聚糖酶, 固态发酵

CLC Number:

S182

YANG Yifan, GAO Yan, LIU Yanan, HUO Xiangdong, LOU Kai, GUAN Bo, CHEN Kaixu, ZENG Jun. Screening of strains producing non-starch polysaccharide enzyme by fermentation of licorice residue and its dynamic changes[J]. Xinjiang Agricultural Sciences, 2024, 61(11): 2733-2741.

杨奕凡, 高雁, 刘亚男, 霍向东, 娄恺, 关波, 陈开旭, 曾军. 甘草渣发酵产非淀粉多糖酶菌株筛选及产酶的动态变化[J]. 新疆农业科学, 2024, 61(11): 2733-2741.

Figures/Tables 13

References 28

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菌株 Strains	D(cm)	d(cm)	D/d	A
黄孢原毛平革 Phanerochaete chrysosporium	3.0±0.063	1.65±0.224	1.818	0.606
康宁木霉 Conning wood mold	3.2±0.141	1.4±0.063	2.286	0.762
黑曲霉 Aspergillus niger	3.0±0.327	1.67±0.122	1.796	0.599
里氏木霉 Trichoderma reeser	2.52±0.043	1.49±0.034	1.691	0.564

菌株 Strains	D(cm)	d(cm)	D/d	A
黄孢原毛平革 Phanerochaete chrysosporium	3.0±0.063	1.65±0.224	1.818	0.606
康宁木霉 Conning wood mold	3.2±0.141	1.4±0.063	2.286	0.762
黑曲霉 Aspergillus niger	3.0±0.327	1.67±0.122	1.796	0.599
里氏木霉 Trichoderma reeser	2.52±0.043	1.49±0.034	1.691	0.564

菌株 Strains	D(cm)	d(cm)	D/d	A
黄孢原毛平革 Phanerochaete chrysosporium	4.37±0.262	1.73±0.205	2.571	0.643
康宁木霉 Conning wood mold	6.1±0.572	1.63±0.125	3.779	0.945
黑曲霉 Aspergillus niger	5.5±0.082	1.47±0.17	4.029	1.007
里氏木霉 Trichoderma reeser	3.97±0.125	1.85±0.15	2.222	0.555

菌株 Strains	D(cm)	d(cm)	D/d	A
黄孢原毛平革 Phanerochaete chrysosporium	4.37±0.262	1.73±0.205	2.571	0.643
康宁木霉 Conning wood mold	6.1±0.572	1.63±0.125	3.779	0.945
黑曲霉 Aspergillus niger	5.5±0.082	1.47±0.17	4.029	1.007
里氏木霉 Trichoderma reeser	3.97±0.125	1.85±0.15	2.222	0.555

培养时间 Incubation time	形态学观察 Morphological observation
第1 d 1st day	培养基底部有少量的白色菌丝
第2 d 2nd day	培养基的表面部分有白色菌丝生成, 黑色孢子逐渐增多
第3 d 3rd day	黑色孢子继续增加, 并且孢子子颜色有所加深
第4 d 4th day	培养基内容物基本变成纯黑色