中药渣中纤维素降解菌的筛选及降解特性

doi:10.13560/j.cnki.biotech.bull.1985.2024-0482

摘要/Abstract

摘要：

目的从中药渣中分离筛选高效纤维素降解菌株，用于中药渣的腐解，提高资源化利用程度。方法采用CMC-Na平板法对纤维素降解菌进行初步筛选，结合刚果红染色法和滤纸条崩解试验进行复筛，通过生理生化及16S rDNA分子生物学进行鉴定，选择不同生长和抗生素培养条件研究菌株纤维素酶活力，并以中药渣为唯一碳源检测菌株的降解能力。结果从中药渣中分离筛选获得3株具有纤维素降解能力的菌株，经鉴定H-1、Z-1为Bacillus sp.，Z-2为Niallia sp.。菌株H-1、Z-1最佳产酶条件为装液量10 mL/50 mL、pH 6、30℃；菌株Z-2最佳产酶条件为装液量10 mL/50 mL、pH 7、37℃，添加土霉素和恩诺沙星对菌株H-1、Z-1、Z-2的酶活力影响不显著。菌株H-1、Z-1和Z-2的中药渣降解率分别为56.37%、54.76%和41.53%，纤维素降解率分别为22.12%、20.01%和9.78%。结论筛选获得的3株菌具有良好的纤维素酶活力，且不受抗生素影响，对中药渣具有较好的降解效果。

关键词: 中药渣, 纤维素降解菌, 降解特性, 产酶条件, 抗生素

Abstract:

Objective To isolate and screen efficient cellulose degrading strains from Chinese herb residues, and they were used to improve resource utilization. Method CMC-Na plate was used to primarily screen cellulose-degrading bacteria, combined Congo red staining and disintegration test of filter paper strips to have re-screening. They were identified by physiology, biochemistry, and 16S rDNA molecular biology. Then the cellulase activities of strains with different growth and antibiotic culture conditions were investigated, and the degrading abilities of strains were determined using Chinese herb residues as the only carbon source. Result Three strains with cellulose-degrading ability were isolated and screened from Chinese herb residues. H-1 and Z-1 were identified as Bacillus sp., and Z-2 as Niallia sp. The optimal enzyme production conditions for strain H-1 and Z-1 were 10 mL/50 mL, pH 6, and 30℃.The optimal enzyme production conditions for strain Z-2 10 mL/50 mL, pH 7, 37℃, and adding oxytetracycline and enrofloxacin did not have a significant effect on the enzyme activities of strain H-1, Z-1 and Z-2. The degradation rates of Chinese herb residues by strain H-1, Z-1 and Z-2 were 56.37%, 54.76% and 41.53%, and the rates of degrading cellulose were 22.12%, 20.01% and 9.78%, respectively. Conclusion The three screened strains have robust cellulase activity, are not affected by antibiotics, and show an effective degradation capability for Chinese herb residues.

Key words: Chinese herb residues, cellulose degrading bacteria, degradation characteristics, enzyme producing conditions, antibiotics

张雨, 周椿富, 彭益萱, 张家豪, 尹心如, 乔策策, 谢越, 汪建飞, 王翔. 中药渣中纤维素降解菌的筛选及降解特性[J]. 生物技术通报, 2025, 41(3): 308-318.

ZHANG Yu, ZHOU Chun-fu, PENG Yi-xuan, ZHANG Jia-hao, YIN Xin-ru, QIAO Ce-ce, XIE Yue, WANG Jian-fei, WANG Xiang. Screening and Degrading Characteristics of Cellulose-degrading Strains in Chinese Herb Residues[J]. Biotechnology Bulletin, 2025, 41(3): 308-318.

图/表 12

图1 菌株产透明圈能力A：CMC-Na平板中菌落情况 B：CMC-Na平板中透明圈情况

Fig. 1 Capacity of strain producing hydrolytic circlesA: Colony condition in CMC-Na plate. B: Hydrolysis circle in CMC-Na plate

表1 菌株的形态结构

Table 1 Morphological structure of strain

菌株编号 Strain No.	形态 Shape	边缘 Edge	颜色 Color	凸起 Hump	湿润 Moist	光滑 Slick	透明 Transparency	光泽 Gloss
H-1	圆形	完整	白色	-	+	+	不透明	+
H-2	不规则	撕裂状	淡黄色	-	+	-	不透明	+
H-3	絮状	不完整	白色	-	-	-	不透明	-
H-4	树枝状	波纹状	白色	-	-	-	不透明	-
H-5	不规则	不完整	乳脂色	+	-	-	不透明	-
Z-1	圆形	完整	白色	-	-	-	不透明	+
Z-2	圆形	完整	白色	-	+	+	半透明	+
Z-3	絮状	不完整	白色	+	-	-	不透明	-
Z-4	圆形	完整	淡粉色	+	+	+	不透明	+
Z-5	不规则	不完整	白色	-	-	-	不透明	-

图2 滤纸崩解情况A：菌株H-1培养3 d滤纸崩解情况；B：菌株Z-1培养3 d滤纸崩解情况；C：菌株Z-2培养3 d滤纸崩解情况；D：菌株H-1培养7 d滤纸崩解情况；E：菌株Z-1培养7 d滤纸崩解情况；F：菌株Z-2培养7 d滤纸崩解情况

Fig. 2 Disintegration of filter paperA: Disintegration of filter paper cultured with strain H-1 for 3 d. B: Disintegration of filter paper cultured with strain Z-1 for 3 d. C: Disintegration of filter paper cultured with strain Z-2 for 3 d. D: Disintegration of filter paper cultured with strain H-1 for 7 d. E: Disintegration of filter paper cultured with strain Z-1 for 7 d. F: Disintegration of filter paper cultured with strain Z-2 for 7 d

图3 菌落形态和革兰氏染色A：菌株H-1在LB上的菌落形态；B：菌株Z-1在LB上的菌落形态；C：菌株Z-2在LB上的菌落形态；D：菌株H-1在光学显微镜中形态（1 000×）；E：菌株Z-1在光学显微镜中形态（1 000×）；F：菌株Z-2在光学显微镜中形态（1 000×）

Fig. 3 Colony morphology and Gram stainingA: Morphology of strain H-1 in LB. B: Morphology of strain Z-1 in LB. C: Morphology of strain Z-2 in LB. D: Morphology of strain H-1 under optical microscope (1 000×). E: Morphology of strain Z-1 under optical microscope (1 000×). F: Morphology of strain Z-2 in light microscope (1 000×)

表2 部分生化特征

Table 2 Biochemical characteristics

项目 Item	结果 Results
项目 Item	H-1	Z-1	Z-2
葡萄糖 Glucose	+	+	+
明胶 Gelatin	+	+	-
运动性 Moveability	+	+	+
硫化氢Hydrothion	-	-	-

图4 菌株基于16S rDNA序列同源性构建的系统发育树

Fig. 4 Phylogenetic tree of strains based on 16S rDNA sequence homology

图5 不同装液量对菌株H-1、Z-1和Z-2酶活力的影响A：不同装液量对菌株滤纸酶活力的影响；B：不同装液量对菌株内切酶活力的影响；C：不同装液量对菌株外切酶活力的影响

Fig. 5 Effects of different liquid loadings on the enzyme activities of strain H-1, Z-1, and Z-2A: Effect of different liquid loading on strain filter paper enzyme activity. B: Effect of different liquid loading on strain incision enzyme activity. C: Effect of different liquid loading on strain excision enzyme activity

图6 不同pH对菌株H-1、Z-1和Z-2酶活力的影响A：pH对菌株滤纸酶活力的影响；B：pH对菌株内切酶活力的影响；C：pH对菌株外切酶活力的影响

Fig. 6 Effects of pH on the enzyme activities of strain H-1, Z-1, and Z-2A: Effect of pH on strain filter paper enzyme activity. B: Effect of pH on strain incision enzyme activity. C: Effect of pH on strain excision enzyme activity

图7 温度对菌株H-1、Z-1和Z-2酶活力的影响A：温度对菌株滤纸酶活力的影响；B：温度对菌株内切酶活力的影响；C：温度对菌株外切酶活力的影响

Fig. 7 Effects of temperatures on the enzyme activities of strain H-1, Z-1, and Z-2A: Effect of different temperature on strain filter paper enzyme activity. B: Effect of temperature on strain incision enzyme activity. C: Effect of temperature on strain excision enzyme activity

图8 不同抗生素对菌株H-1、Z-1和Z-2酶活力的影响A：不同抗生素对菌株滤纸酶活力的影响；B：不同抗生素对菌株内切酶活力的影响；C：不同抗生素对菌株外切酶活力的影响，不同字母代表差异显著（P<0.05）

Fig. 8 Effects of different antibiotics on the enzyme activities of strain H-1, Z-1, and Z-2A: Effect of adding different antibiotics on strain filter paper enzyme activity. B: Effect of adding different antibiotics on strain incision enzyme activity. C: Effect of adding different antibiotics on strain excision enzyme activity, Different letters indicate significant differences (P<0.05)

图9 接种菌株H-1、Z-1和Z-2对中药渣降解率的影响

Fig. 9 Effects of inoculated strain H-1, Z-1 and Z-2 on the degradation rates of Chinese herb residues

图10 接种菌株H-1、Z-1和Z-2对中药渣纤维素组分的影响

Fig. 10 Effects of inoculated strain H-1, Z-1, and Z-2 on the cellulose components of Chinese herb residues

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