Screening of a lignin degrading bacterium and its degradation pathway

doi:10.16085/j.issn.1000-6613.2020-1541

Abstract

Abstract:

The content of lignin and alkaline salt in the pretreatment solution of corn straw obtained by high temperature alkaline salt treatment was high. In order to reuse the pretreatment solution containing salt, soluble lignin should be removed. In this experiment, a lignin-degrading strain CCZU-WJ6 was screened and isolated from rotten wood and was identified as Achromobacter sp. by morphological observation and 16S rDNA sequence analysis. The activities of enzymes related to lignin degradation in CCZU-WJ6, the growth trend of CCZU-WJ6 in corn straw pretreatment solution, COD removal rate and lignin degradation rate of the pretreatment solution were measured. GC-MS and FTIR were used to detect the changes of components and chemical bonds in the pretreatment solution. Through single-factor experiment, the optimal conditions for degradation of the pretreatment solution by CCZU-WJ6 were determined as pH 7.0, temperature 30℃, dilution factor 10 times, and inoculation amount 0.5g/mL. After 6 days of treatment, the removal rate of COD and the degradation rate of lignin were 40.9% and 32.1%, respectively. The activities of lignin peroxidase and manganese peroxidase secreted by CCZU-WJ6 were 215.5U/L and 200.1U/L, respectively. CCZU-WJ6 can destroy the benzene ring structure, ether bond and C＝O bond in the lignin structure, and the degradation products contained guaiacol and p-coumaric acid, which inferred that the degradation pathways are β-aryl ether metabolic pathway and ferulic acid metabolic pathway. CCZU-WJ6 can be used to degrade industrial wastewater containing lignin and has broad prospects in industrial applications.

Key words: lignin, degradation pathway, lignin peroxidase, manganese peroxidase

摘要：

经高温碱性盐法预处理的玉米秸秆预处理液中木质素和碱性盐含量高，为了反复利用含盐预处理液，需脱除其中的可溶性木质素。本实验从腐木中筛选分离到一株木质素降解菌株CCZU-WJ6，通过形态学观察和16S rDNA序列分析，鉴定该菌株属于无色杆菌属（Achromobacter sp.）。测定了CCZU-WJ6木质素降解相关酶的活性，及其在玉米秸秆预处理液中的生长趋势、预处理液的COD去除率和木质素降解率，通过GC-MS和FTIR检测预处理液中组分及其化学键的变化。通过单因素实验确定了菌株CCZU-WJ6处理预处理液的最佳条件为pH 7.0、温度30℃、预处理液稀释倍数10倍、接菌量0.5g/mL。在处理6天后，预处理液COD脱除率为40.9%，木质素的降解率为32.1%，CCZU-WJ6分泌木质素过氧化物酶和锰过氧化物酶的酶活分别达到215.5U/L、200.1U/L。CCZU-WJ6会破坏木质素结构中的苯环结构、醚键以及C＝O键，降解产物中含有愈创木酚和对香豆酸，推断其降解途径为β-芳基醚代谢途径和阿魏酸代谢途径。CCZU-WJ6可用来处理含木质素的工业废水，在工业方面具有广阔的应用前景。

关键词: 木质素, 降解途径, 木质素过氧化物酶, 锰过氧化物酶

CLC Number:

X703

WANG Jing, NI Jinying, WANG Liqun, QING Qing, YAN Shenghu, ZHANG Yue. Screening of a lignin degrading bacterium and its degradation pathway[J]. Chemical Industry and Engineering Progress, 2021, 40(7): 4021-4026.

王晶, 倪金荧, 王利群, 卿青, 严生虎, 张跃. 一株木质素降解细菌的筛选及其降解途径[J]. 化工进展, 2021, 40(7): 4021-4026.

Figures/Tables 8

References 13

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编号	保留时间/min	名称	结构式	降解时间/d
编号	保留时间/min	名称	结构式	0	3	6
A1	11.589	三苯基磷	C₁₈H₁₆P	+	-	-
A2	13.684	三苯基磷胺	C₁₈H₁₆NP	+	-	-
A3	14.527	三苯基氧化磷	C₁₈H₁₅OP	+	-	-
B1	9.008	愈创木酚	C₇H₈O₂	-	+	+
B2	10.082	对香豆酸	C₉H₈O₃	-	+	+

编号	保留时间/min	名称	结构式	降解时间/d
编号	保留时间/min	名称	结构式	0	3	6
A1	11.589	三苯基磷	C₁₈H₁₆P	+	-	-
A2	13.684	三苯基磷胺	C₁₈H₁₆NP	+	-	-
A3	14.527	三苯基氧化磷	C₁₈H₁₅OP	+	-	-
B1	9.008	愈创木酚	C₇H₈O₂	-	+	+
B2	10.082	对香豆酸	C₉H₈O₃	-	+	+

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