木质素氧化酶系高产复合真菌培养体系构建

doi:10.11707/j.1001-7488.LYKX20230074

Abstract

Abstract:

Objective: This study aims to obtain the optimal ligninolytic enzymes production system through the preliminary screening of fungal strains, determination of the optimal culture conditions and construction of the compound fungal culture system, which provides basic and scientific basis for pretreatment of bio-pulping. Method: The fungal fruiting bodies on the trees of the Summer Palace were collected as the source of strains. The white rot fungi strains with fast growth rate and high laccase activity were screened by growth rate and guaiacol culture medium. Then, the secretion of laccase (Lac), manganese- peroxidase (MnP) and lignin-peroxidase (LiP) were measured. The effects of different carbon sources, nitrogen sources, pH, additives, metal ions, temperature on the secretion of ligninolytic enzyme system were studied by single factor experiment. According to the enzyme production, different strains were selected to construct the 1∶1 compound fungal culture system, and the enzyme production and enzyme production efficiency were studied. Result: A strain with rapid growth and high laccase activity was obtained and identified as white rot fungus Trametes hirsute. The results of single factor test showed that glucose and yeast extract were the optimal carbon and nitrogen sources for T. hirsute ligninolytic enzymes production, respectively. The ligninolytic enzymes production efficiency of adding Cu^{2 +} alone was better than that of adding both Cu²⁺ and Mn²⁺. The ligninolytic enzymes production increased first and then decreased with the increase of metal ion concentration, and 0.25 mmol·L^?1 Cu^{2 +} was the best. At 38 ℃, pH=6.5 and 0.5 mmol·L^?1 veratrol and 0.5 mmol·L^?1 p-phenylenediamine, the lignin oxidase production of T. hirsute was promoted. Based on the optimization of culture conditions, a composite fungal culture system of T. hirsute and Gloeophyllum trabeum was constructed, and its enzyme production ability was further improved. The maximum total lignin oxidase production of the cultures reached 1 505.81 U·L^?1, which was about 63.61% higher than that of the control group. The laccase activity was 84.37% higher than that of the control group, and total ligninolytic enzymes production peak was reached one day in advance. Conclusion: High laccase activity fungus T. hirsute was obtained by preliminary screening. Its ligninolytic enzyme system was significantly increased by optimizing the enzyme production conditions. Based on the optimized medium, the T. hirsute and G. trabeum compound fungal culture system further improved the total lignin oxidase production and promoted laccase to reach the peak enzyme production one day in advance. The research provides a basis for the efficient utilization of lignocellulose with biological pretreatment.

Key words: Trametes hirsute, ligninolytic enzyme system, compound fungal culture system, biological pretreatment, laccase

CLC Number:

S782.33

Yixin Xiong,Shuang Wang,Xingxia Ma,Zhiqin Sun. Construction of Compound Fungal Culture System for High-Yield Ligninolytic Enzymes[J]. Scientia Silvae Sinicae, 2024, 60(10): 133-142.

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试验组 Test	Cu²⁺ 0.25 mmol·L^?1	VA 0.5 mmol·L^?1	对苯二胺 P-phenylenediamine 0.5 mmol·L^?1	复合培养 Compound culture	Lac/ (U·L^?1)	MnP/ (U·L^?1)	LiP/ (U·L^?1)	总酶活性 Total enzyme activity/ (U·L^?1)
对照组Control	—	—	—	—	608.21	122.92	189.22	920.35
1	+	+	—	—	788.29	131.49	227.65	1 147.43
2	+	—	+	—	897.85	126.30	216.76	1 240.91
3	+	+	+	—	966.50	114.58	234.65	1 315.73
4	+	+	—	+	1 043.08	134.65	246.59	1 424.32
5	+	—	+	+	1 114.71	129.25	241.84	1 485.80
6	+	+	+	+	1 121.32	124.81	259.68	1 505.81

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Construction of Compound Fungal Culture System for High-Yield Ligninolytic Enzymes

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