双叉犀金龟肠道纤维素降解菌复合菌系的构建、产纤维素酶条件优化及菌糠降解能力评价

doi:10.16380/j.kcxb.2025.09.007

昆虫学报 ›› 2025, Vol. 68 ›› Issue (9): 1233-1241.doi: 10.16380/j.kcxb.2025.09.007

双叉犀金龟肠道纤维素降解菌复合菌系的构建、产纤维素酶条件优化及菌糠降解能力评价

朱恋, 贾金姗, 李森多, 吴钧浩, 何奕俊, 张俊杰, 杜文梅^*, 胡莹^*

(吉林农业大学生物防治研究所, 吉林省农业害虫生物防治国际科技合作重点实验室, 长春 130118)

出版日期:2024-09-20 发布日期:2025-10-28

Optimization of cellulase production conditions of composite bacterial consortia constructed with the gut-derived cellulose-degrading bacteria from Allomyrina dichotomus (Coleoptera: Scarabaeidae) and assessment of their degradation efficiency for spent mushroom substrate

ZHU Lian, JIA Jin-Shan, LI Sen-Duo, WU Jun-Hao, HE Yi-Jun, ZHANG Jun-Jie, DU Wen-Mei^*, HU Ying^*

(Jilin Provincial International Cooperation Key Laboratory for Biological Control of Agricultural Pests, Institute of Biological Control, Jilin Agricultural University, Changchun 130118, China)

Online:2024-09-20 Published:2025-10-28

摘要/Abstract

摘要： 【目的】筛选双叉犀金龟Allomyrina dichotomus肠道纤维素降解菌株构建复合菌系，研究其菌糠降解能力，为废弃菌糠的处理提供理论基础和实践依据。【方法】利用双叉犀金龟幼虫肠道分离纯化得到的6株较高酶活力的纤维素降解细菌(贝莱斯芽孢杆菌Bacillus velezensis M24, 枯草芽孢杆菌Bacillus subtilis H12, H11, H4和M33及暹罗芽孢杆菌Bacillus siamensis M32)，以互不拮抗为前提构建复合菌系，测定其4种纤维素酶(滤纸酶、内切葡聚糖苷酶、外切葡聚糖苷酶和β葡萄糖苷酶)活力，依据酶活力谱选定最优复合菌系，对复合菌系的培养时间、接种量和培养温度进行优化，并通过差重法对降解前后菌糠结构变化进行扫描电子显微镜观察。【结果】基于滤纸酶、内切葡聚糖苷酶、外切葡聚糖苷酶和β-葡萄糖苷酶的活力(分别为18.08, 69.37, 19.09和17.95 U/mL)，构建复合菌系M24∶H11； M24∶H11最佳培养时间2 d、接菌量1%和培养温度40 ℃，优化后4种纤维素酶活力显著提升1.98～2.16倍； 25 ℃下30 d内， M24∶H11和单菌株菌糠降解率随时间均呈显著上升趋势， M24∶H11菌糠降解率显著高于单菌株，最高可达38.04%； M24∶H11的降解处理有效地打开了菌糠表面结构，接触面积增大。【结论】本研究构建了双叉犀金龟幼虫肠道来源的复合菌系M24∶H11，对其进行产酶条件优化。M24∶H11可有效降解菌糠，为食用菌废弃物的高效处理提供了新的技术思路和理论依据。

关键词: 双叉犀金龟, 肠道微生物, 复合菌系, 纤维素酶, 酶活力, 菌糠

Abstract: 【Aim】To screen cellulose-degrading bacterial strains from the gut of Allomyrina dichotomus for constructing composite bacterial consortia and to investigate their degradation capacity for spent mushroom substrate (SMS), thereby providing theoretical and practical foundations for SMS waste management.【Methods】Composite bacterial consortia were constructed based on non-antagonistic relationships among six cellulose-degrading bacteria with high enzyme activity (Bacillus velezensis M24, Bacillus subtilis H12, H11, H4 and M33, and Bacillus siamensis M32) isolated and purified from the larval gut of A. dichotomus. The activities of four cellulases (filter paper enzyme, endoglucanase, exoglucanase and β-glucosidase) were determined, the optimal composite bacterial consortia was selected according to its enzymatic activity profile, and the culture time, inoculation amount and incubation temperature for the selected composite bacterial consortia were optimized. The structural changes in the spent mushroom substrate before and after degradation were observed through scanning electron microscope by differential weight method.【Results】The composite bacterial consortia M24∶H11 was constructed based on the enzymatic activities of filter paper enzyme, endoglucanase, exoglucanase and β-glucosidase (18.08, 69.37, 19.09 and 17.95 U/mL, respectively). The optimal culture time, inoculation amount and culture temperature of M24∶H11 were 2 d, 1% and 40 ℃, respectively. Following optimization, the activities of the four cellulases increased significantly by 1.98-2.16-fold. At 25 ℃ within 30 d, M24∶H11 showed a significantly higher SMS degradation rate (up to 38.04%) than individual strains, with the degradation rate increasing over time. The degradation by M24∶H11 effectively disrupted the SMS surface structure, increasing its contact area.【Conclusion】The larval gut-derived composite bacterial consortia M24∶H11 of A. dichotomus was constructed and optimized for enzyme production, with high efficiency in SMS degradation. This study offers novel technical insights and theoretical support for the efficient control of the edible fungi waste.

Key words: Allomyrina dichotomus, gut microorganisms, composite bacterial consortia, cellulase, enzyme activity, spent mushroom substrate

朱恋, 贾金姗, 李森多, 吴钧浩, 何奕俊, 张俊杰, 杜文梅, 胡莹. 双叉犀金龟肠道纤维素降解菌复合菌系的构建、产纤维素酶条件优化及菌糠降解能力评价[J]. 昆虫学报, 2025, 68(9): 1233-1241.

ZHU Lian, JIA Jin-Shan, LI Sen-Duo, WU Jun-Hao, HE Yi-Jun, ZHANG Jun-Jie, DU Wen-Mei, HU Ying. Optimization of cellulase production conditions of composite bacterial consortia constructed with the gut-derived cellulose-degrading bacteria from Allomyrina dichotomus (Coleoptera: Scarabaeidae) and assessment of their degradation efficiency for spent mushroom substrate[J]. Acta Entomologica Sinica, 2025, 68(9): 1233-1241.

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双叉犀金龟肠道纤维素降解菌复合菌系的构建、产纤维素酶条件优化及菌糠降解能力评价

Optimization of cellulase production conditions of composite bacterial consortia constructed with the gut-derived cellulose-degrading bacteria from Allomyrina dichotomus (Coleoptera: Scarabaeidae) and assessment of their degradation efficiency for spent mushroom substrate

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