Screening and Degrading Characteristics of Cellulose-degrading Strains in Chinese Herb Residues

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

Abstract

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

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.

Figures/Tables 12

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

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	-

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

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×)

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	-	-	-

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

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

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

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

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)

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

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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