Optimization of Indole-degrading Conditions in Pig Manure Waste Water by Enteroccus hirae IDO5 and Analysis of Its Corresponding Degradation Pathway

doi:10.13560/j.cnki.biotech.bull.1985.2021-0071

Abstract

Abstract:

A highly efficient indole-degrading bacterium strain IDO5 was identified in the present study. Herein,the indole-degrading ability of IDO5 in pig manure waste water was mainly investigated. Firstly,IDO5 was identified by morphology and 16S rRNA sequence analysis,and the degrading effect of IDO5 on indole and the types of substrates were preliminary analyzed. Secondly,the effects of rotational speed,temperature,pH and carbon source on the degradation of indole in pig manure waste water by IDO5 were compared. Finally,the structure and degradation pathway of indole degraded by IDO5 were analyzed by GC-MS. The results showed that IDO5 could not only use indole as the carbon source for growth,but also degrade 3-methylindole,phenol,cresol and other pollutants,thus IDO5 had a broad substrate spectrum. The optimal conditions for IDO5 degrading indole in the pig manure were as:temperature 37^oC,pH 9 and rotary speed 170 r/min. Under these conditions,98.2% of indole in the pig manure wastewater was degraded within 24 h,i.e.,the removal rate was quite high. IDO5 removed 93.7% indole even when an additional 100 mg/L indole was added. IDO5 formed a red product during the degradation of indole. The degradation of indole by strain IDO5 may be through the pathway of indole → isatin → anthranilic acid → 2-methylbenzalde after analyzed by GC-MS. Strain IDO5 not only has strong indole-degrading ability in a wide pH range,but also can act on a variety of indole derivatives and other compounds. Therefore,strain IDO5 can effectively degrade indole in livestock and poultry breeding waste water under a wide range of conditions,and has certain application value.

Key words: Enterococcus hirae, indole, pH stability, aquaculture waste water

YU Qin, MA Xian-yong, DENG Dun, WANG Yong-fei. Optimization of Indole-degrading Conditions in Pig Manure Waste Water by Enteroccus hirae IDO5 and Analysis of Its Corresponding Degradation Pathway[J]. Biotechnology Bulletin, 2021, 37(12): 113-123.

Figures/Tables 13

Fig. 1 Morphological characteristics of IDO5 A:Plate colony of IDO5 strain;B:scanning electron microscopy result of strain IDO5

Fig. 2 Phylogenetic tree based on 16S rRNA sequence of IDO5 Phylogenetic neighbor joining tree of IDO5 is constructed based on 16S rRNA gene sequences. The phylogenetic tree shows the relationships between individual isolates and species within the respective genus. Bootstrap values（expressed as percentages of 1 000 replications）given at nodes are > 50%. Bar 0.05% sequence variation. GenBank accession numbers are given in parentheses

Fig. 3 HPLC results of treated indole by IDO5 bacterial solution and control

Table 1 Substrate spectrum of strain IDO5

Fig. 4 Effects of temperature on the degradation of indole in pig anure wastewater by IDO5 Error bars indicate the standard deviation and different letters reflect significant difference（P< 0.05）between treatments. The same below

Fig. 5 Effect of rotating speed on the degradation of indole in pig manure wastewater by strain IDO5

Fig. 6 Effect of pH on the degradation of indole in pig manure wastewater by strain IDO5

Fig. 7 Effect of exogenous carbon source on the degrada-tion of indole in pig manure wastewater by strain IDO5

Fig. 8 Variation of indole degradation in pig manure water by strain IDO5 during 24 h

Fig.9 Variation of indole degradation in different concentrations by strain IDO5 during 24 h

Table 2 Target component spectrum of indole degrading GC-MS by strain IDO5

Table 3 Resources of indole degrading bacteria

菌株名称 Strain name	降解特性 Degradation characteristics	参考文献 Reference
Pseudomonas aeruginosaGs	36 d时降解58.5-175 mg/L吲哚 58.5-175 mg/L indole was degraded after 36 d	[25]
Comamonas sp. IDO1	至少需要160 h降解100 mg/L吲哚 100 mg/L indole was degraded after at least 160 h	[26]
Xenophilus sp. IDO4	至少需要160 h降解100 mg/L吲哚 100 mg/L indole was degraded after at least 160 h	[26]
Arthrobacter sp. B1	40 h降解117 mg/L吲哚 117 mg/L indole was degraded after 40 h	[27]
Arthrobacter sp. SPG	36 h降解58.5 mg/L吲哚 58.5 mg/L indole was degraded after 46 h	[28]
Providencia sp.	28 h降解100 mg/L吲哚 100 mg/L indole was degraded after 28 h	[29]
Cupriavidus sp. SHE	24 h降解100 mg/L吲哚 100 mg/L indole was degraded after 24 h	[23]

Fig.10 Pathway of indole degradation by strain IDO5

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