Isolation and Identification of Nicosulfuron Degrading Strain Chryseobacterium sp. LAM-M5 and Study on Its Degradation Pathway

doi:10.13560/j.cnki.biotech.bull.1985.2021-0243

Abstract

Abstract:

In order to obtain more nicosulfuron-degrading bacterial resources,the activated sludge samples from a nicosulfuron manufacturer in Hefei city were used for study object,from which an isolated and screened strain grew well with glucose as sole carbon source and nicosulfuron as sole nitrogen source. Based on its phenotypic characteristics,the 16S rRNA gene sequence similarity,the values of DNA-DNA hybridization and average nucleotide identity,it was identified as Chryseobacterium lacus LAM-M5. This strain degraded 92.39% of 50 mg/L nicosulfuron within 7 d. Liquid chromatography mass spectrometry was used to detect and identify the metabolites after mixing culture of LAM-M5 and nicosulfuron,and 5 major substances were detected. The metabolic pathway of LAM-M5 degrading nicosulfuron was primarily speculated based on the chemical structure and characteristics of intermediates. High performance liquid chromatography was employed to isolate and identify the acids generated during the degradation of nicosulfuron by strain LAM-M5,and the results showed that L-malic acid was predominant. It was deduced that strain LAM-M5 metabolized the glucose into L-malic acid when its growth was stressed by the nicosulfuron,from which the environmental pH decreased and the nicosulfuron hydrolyzed,thus the stress was released. Through the analysis of the whole genome sequence of strain LAM-M5,7 annotated genes that may contribute to the degradation of nicosulfuron were predicted. The results reveal that strain LAM-M5 has excellent ability of degrading nicosulfuron and may have promising application potential in the microbial remediation of nicosulfuron -polluted environment.

Key words: nicosulfuron, Chrysobacterium, microbial degradation, metabolic pathway

MA Qing-yun, JIANG Xu, LI Qing-qing, SONG Jin-long, ZHOU Yi-qing, RUAN Zhi-yong. Isolation and Identification of Nicosulfuron Degrading Strain Chryseobacterium sp. LAM-M5 and Study on Its Degradation Pathway[J]. Biotechnology Bulletin, 2022, 38(2): 113-122.

Figures/Tables 8

Fig.1 Chemical structural formula of nicosulfuron

Fig.2 Colony morphology of strain LAM-M5 on the beef extract peptone medium plate

Fig.3 Neighbour-joining phylogenetic tree based on the 16S rRNA gene sequences

Fig.4 Effects of different pH（A）,temperature（B）and initial inoculum ratio（C）on the degradation of nicosulfuron by strain LAM-M5 The error line in the figurebars refers to standard errors. The same below

Fig.5 Change of pH value during the nicosulfuron degra-dation by strain LAM-M5

Fig.6 Proposed metabolic pathways of nicosulfuron degra-dation by strain LAM-M5

Table 1 Genomic characteristics of strain LAM-M5

基因组组分 Characteristics	菌株LAM-M5 Chryseobacterium lacus LAM-M5
Genome size/bp	6 311 696
Gene number	6 366
Gene length/bp	5 704 104
GC content/%	43.84
genes of genome/%	90.37
Gene average length/bp	896
Gene internal length/bp	607 592
Gene internal GC content/%	36.52
Internal of genome/%	9.63

Table 2 7 genes related to degradation enzymes in the genome of strain LAM-M5

编号 No.	预测的降解酶基因 Predicted degeration genes	大小 Size/bp
1	Putative phosphoesterase	172
2	Acyl-CoA thioesterase YneP	264
3	Carboxylesterase	254
4	Catalase X	535
5	Probable manganese catalase	274
6	Hydrolase YhcX	513
7	Putative sporulation hydrolase CotR	317

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