The Key Laboratory of Feed Biotechnology of Ministry of Agriculture, National Engineering Research Center of Biological Feed, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing, China.
Department of Biology, Faculty of Natural Sciences, Mettu University, Mettu, Ethiopia.
PLoS One. 2019 Jan 28;14(1):e0211275. doi: 10.1371/journal.pone.0211275. eCollection 2019.
3-Methylindole (3MI) or Skatole is a volatile lipophilic organic compound produced by anoxic metabolism of L-tryptophan and associated with animal farming and industrial processing wastes. Pure cultures of bacteria capable of utilizing 3MI were isolated from chicken manure using enrichment culture techniques. The bacteria were identified as Acinetobacter toweneri NTA1-2A and Acinetobacter guillouiae TAT1-6A, based on 16S rDNA gene amplicon sequence data. The optimal temperature and pH for degradation of 3MI were established using single factor experiments. Strain tolerance was assessed over a range of initial concentrations of 3MI, and the effects of initial concentration on subsequent microbial 3MI degradation were also measured. During the degradation experiment, concentrations of 3MI were quantified by reverse-phase high-performance liquid chromatography (HPLC). The strains were capable of degrade initial concentrations of 3MI ranging from 65-200 mg/L. The degradation efficiency was >85% in 6 days for both strains when the initial concentration is less than 200 mg/L. The strains were tested for enzymatic activity using 65 mg/L 3MI. The enzyme extracts of NTA1-2A and TAT1-6A from the 3MI medium degraded 71.46% and 60.71% of 3MI respectively, but no appreciable change in 3MI concentration in the control group was witnessed. Our experiment revealed betaine and choline were identified as 3MI degradation metabolites by both strains while nitroso-pyrrolidine and beta-alaninebetaine formed by NTA1-2A and TAT1-6A strains respectively. The NTA1-2A and TAT1-6A strains removed 84.32% and 81.39% 3MI respectively from chicken manure during fermentation in 8 days and showed a statistically significant difference (P < 0.05) compared with the control group. The optimum temperature and pH were 31°C and 6 respectively, for 3MI degradation by A. toweneri NTA1-2A and A. guillouiae TAT1-6A. We concluded that A. toweneri NTA1-2A and A. guillouiae TAT1-6A are potential strains of interest to degrade 3MI and control odorant in poultry and other livestock industries.
3-甲基吲哚(3MI)或粪臭素是一种挥发性亲脂有机化合物,由缺氧代谢 L-色氨酸产生,与动物养殖和工业加工废物有关。使用富集培养技术从鸡粪中分离出能够利用 3MI 的纯细菌培养物。根据 16S rDNA 基因扩增子序列数据,这些细菌被鉴定为不动杆菌toweneri NTA1-2A 和不动杆菌guillouiae TAT1-6A。通过单因素实验确定了 3MI 降解的最佳温度和 pH 值。通过一系列 3MI 初始浓度评估了菌株的耐受性,并测量了初始浓度对随后微生物 3MI 降解的影响。在降解实验中,通过反相高效液相色谱(HPLC)定量 3MI 浓度。两种菌株均能够降解 65-200mg/L 的初始 3MI 浓度。当初始浓度小于 200mg/L 时,两种菌株在 6 天内的降解效率均大于 85%。使用 65mg/L 3MI 测试了菌株的酶活性。从 3MI 培养基中提取的 NTA1-2A 和 TAT1-6A 酶提取物分别降解了 71.46%和 60.71%的 3MI,但对照组中 3MI 浓度没有明显变化。我们的实验表明,两种菌株均将甜菜碱和胆碱鉴定为 3MI 降解代谢物,而 NTA1-2A 和 TAT1-6A 菌株分别形成亚硝基吡咯烷和β-丙氨酸甜菜碱。在 8 天的发酵过程中,NTA1-2A 和 TAT1-6A 菌株分别从鸡粪中去除了 84.32%和 81.39%的 3MI,与对照组相比具有统计学意义(P<0.05)。对于 A. toweneri NTA1-2A 和 A. guillouiae TAT1-6A 降解 3MI 的最佳温度和 pH 值分别为 31°C 和 6。我们得出结论,A. toweneri NTA1-2A 和 A. guillouiae TAT1-6A 是潜在的菌株,可用于降解家禽和其他畜牧业中的 3MI 和控制气味。