贵州农业科学2025，Vol.53Issue(7)：68-80,13.DOI:10.3969/j.issn.1001-3601.2025.07.008
中药渣中木质素高效降解菌株的筛选与鉴定及培养条件优化
Filter and Identification of the Strains with Efficient Degradation Ability of Lignin in Chinese Medicine Residue and the Optimization of Culture Conditions
摘要
Abstract
[Objective]The microbial strains with efficient degradation ability of lignin in Chinese herbal medicine residues were screened to clarify the cultivation conditions,thereby providing reference for enhancing the biodegradation efficiency of herbal residues and promoting the resource utilization.[Method]The lignin degrading bacteria was separated and screened from traditional Chinese medicine residue by using aniline blue screening plate combined with degradation test of calcium lignosulfonate,and the lignin degrading bacteria was identified through morphological features,physiological and biochemical characteristics and multi gene molecular systematics.Using the single factor method,media were prepared with varying carbon sources,nitrogen sources,inorganic salts and trace element additives to determine the biomass and sporulation rate of highly efficient lignin-degrading strains.Based on the optimized medium as the basal medium,the viable cell count of the efficient lignin-degrading strain was determined under different cultivation conditions including pH,incubation temperature,shaking speed,medium-to-flask ratio,and inoculum size.[Result]A total of 15 bacterial strains were isolated from Chinese herbal residues using aniline blue lignin-degrading screening medium.During primary screening,strains exhibiting relative enzyme production rate≥6.5 after 24-hour culture were YZ2023-b3,YZ2023-b4,YZ2023-b7,YZ2023-b8,YZ2023-b11 and YZ2023-b13,with YZ2023-b4 showing the highest activity.Strains exhibiting relative enzyme production rate≥7.5 after 48-hour culture were YZ2023-b3,YZ2023-b4,YZ2023-b7,YZ2023-b11 and YZ2023-b13,with YZ2023-b3 demonstrating the highest activity.During the secondary screening phase,strain YZ2023-b4 demonstrated optimal degradation performance with calcium ligno sulfonate of 0.59 mg/mL,followed by YZ2023-b3(0.75 mg/mL),whereas the remaining three strains exhibited residual calcium ligno sulfonate concentration＞1.00 mg/mL.According to morphological features,physiological and biochemical characteristics and multi gene molecular systematics,strain YZ2023-b3 was identified as B.aryabhattai,and strain YZ2023-b4 was identified as B.megaterium.As for the culture media,YZ2023-b3 had the highest biomass content in media with potato starch as the carbon resource,with OD600 absorbance value of 1.46,followed by the one in media with soluble starch as the carbon resource,with OD600 absorbance value of 1.45.YZ2023-b4 had the highest biomass content in media with soluble starch as the carbon resource,with OD600 absorbance value of 1.46.The two strains had both the highest biomass content in media with soybean meal as the nitrogen resource,with OD600 absorbance value of 1.98 and 2.15,respectively.The two strains had both the highest biomass content in media added with MgCl2,with OD600 absorbance value of 1.82 and 1.70,respectively.When the concentration of ZnCl2 was 6 mg/mL,the two strains obtained the highest sporulation rate with 86.63％and 84.00％,respectively.As for culture condition,strains YZ2023-b3 and YZ2023-b4 demonstrated most effective viable cells at specific conditions:pH 7 produced values of 83.40× 108 CFU/mL of YZ2023-b3 and 95.13× 108 CFU/mL of YZ2023-b4,respectively;28℃ produced 82.60× 108 CFU/mL and 94.60× 108 CFU/mL,respectively;220 r/min shaking speed generated 102.40× 108 CFU/mL and 108.70× 108 CFU/mL,respectively;5％inoculation ratio achieved 136.80× 108 CFU/mL and 138.40 × 108 CFU/mL,respectively;200 mL volume(40％flask capacity)resulted in 124.90 × 108 CFU/mL and 135.20× 108 CFU/mL.[Conclusion]The strains B.aryabhattai and B.megaterium have good potential for application in lignin degradation.The optimal culture media includes 5 g/L soluble starch,10 g/L soybean meal,10 g/L MgCl2 and 6 mg/L ZnCl2.And the optimal culture conditions are initial pH at 7,culture temperature at 28 ℃,medium-to-flask ratio at 40％,inoculation rate at 5％and shaking speed at 220 r/min.关键词
中药渣/木质素/微生物降解/培养条件/产酶率Key words
Chinese medicine residue/lignin/microbial degradation/culture condition/production rate of enzyme分类
农业科技引用本文复制引用
宋莲钗,顾小凤,芶久兰,曾祥琼,柳玲玲..中药渣中木质素高效降解菌株的筛选与鉴定及培养条件优化[J].贵州农业科学,2025,53(7):68-80,13.基金项目
贵州省农业科学院博士基金项目"低温纤维素降解菌筛选、酶学特征及产酶条件优化研究"(黔农科青年基金[2023]09号) （黔农科青年基金[2023]09号）
贵州省科技创新平台建设项目"贵州省农业微生物全省重点实验室"(黔科合平台[2025]029) （黔科合平台[2025]029）
贵州省科技支撑计划项目"基于'气流膜'发酵的牛粪+中药渣共堆肥关键技术研究"(黔科合[2023]一般467) （黔科合[2023]一般467）
贵州省农业科学院"揭榜挂帅"项目"农业生物质多元化利用核心技术研究与应用"(黔农科JBGS[2024]05号) （黔农科JBGS[2024]05号）
