海洋源β-内酰胺类抗生素降解菌的分离鉴定及全基因组测序分析

doi:10.16431/j.cnki.1671-7236.2026.02.042

摘要/Abstract

摘要：

目的分离并筛选海洋源β-内酰胺类抗生素降解细菌，分析其降解活性和全基因组，为修复环境中β-内酰胺类抗生素污染提供新的思路。方法采集湛江地区的海水，使用含有青霉素钾、阿莫西林、头孢曲松钠的选择培养基分离和筛选海水中的细菌，通过快速生物降解试验测定分离菌的降解率，对具有降解活性的菌株进行分子生物学鉴定并绘制时间-降解曲线，选择在120 h内降解超过90%抗生素的降解菌，分析其在更高初始抗生素浓度下的降解效果，筛选降解效果最好的1株细菌进行全基因组测序分析。结果从湛江地区的海水中筛选出18株β-内酰胺类抗生素耐药菌，其中5株细菌能快速降解β-内酰胺类抗生素。菌株DA02和DA03能在120 h内降解99.12%和91.21%初始浓度为100 mg/L的头孢曲松钠，且DA02能快速降解初始浓度为300 mg/L的头孢曲松钠。16S rRNA基因序列比对分析结果显示，菌株DA02为嗜麦芽窄食单胞菌。全基因组测序鉴定菌株DA02基因组大小为4 786 356 bp，GC含量为66.58%，共预测到4 253个编码基因，73个tRNAs和7个rRNAs，发现多个编码β-内酰胺酶的基因，包括L1_BLA、LRA-18、NmcR、PSV-1、SRT-1，且该菌含有多种抗生素耐药基因，如AAC（6'）、APH（3'）、cmlv和arr-2，以及丰富的碳水化合物代谢途径基因，参与碳代谢、硫代谢和氮代谢的基因分别有91、12和13个。结论海水中存在高效的β-内酰胺类抗生素降解菌，本研究全基因组分析揭示了头孢曲松钠降解菌DA02的可能降解机制和生物修复潜力，为修复环境中β-内酰胺类抗生素污染提供了潜在的有效方法。

关键词: 生物降解; 海水; β-内酰胺类抗生素; 全基因组

Abstract:

Objective This study aimed to isolate and screen β-lactam antibiotic-degrading bacteria，analyze their degradation activity and whole genome， and provide new ideas for repairing β-lactam antibiotic pollution in the environment. Method In this study， selective media containing penicillin potassium， amoxicillin， and ceftriaxone sodium were used to screen bacteria from seawater in Zhanjiang city in Guangdong province. The degradation rates were determined through rapid biodegradation tests. Molecular biological identification was performed on strains with degradation activity， and time-degradation curves were drawn. Degrading bacteria that degraded more than 90% of antibiotics within 120 h were selected to analyze the degradation effect at higher initial antibiotic concentrations， and the strain with the best degradation effect was screened for whole genome sequencing analysis. Result 18 strains of β-lactam antibiotic-resistant bacteria were screened from seawater in Zhanjiang， among which 5 strains could rapidly degrade β-lactam antibiotics. By drawing time-degradation curves， it was found that strains DA02 and DA03 could degrade 99.12% and 91.21% of ceftriaxone sodium with an initial concentration of 100 mg/L within 120 h， and DA02 could rapidly degrade ceftriaxone sodium with an initial concentration of 300 mg/L. The 16S rRNA gene sequencing analysis results showed that strain DA02 was Stenotrophomonas maltophilia. Whole genome sequencing identified strain DA02 with a genome size of 4 786 356 bp， a GC content of 66.58%， and a total of 4 253 predicted coding genes， 73 tRNAs， and 7 rRNAs. Multiple genes encoding β-lactamases were found， including L1_BLA， LRA-18， NmcR， PSV-1 and SRT-1. The bacterium also contained various antibiotic resistance genes， such as AAC（6'）， APH（3'）， cmlv and arr-2. It also possessed abundant carbohydrate metabolic pathways， with 91， 12 and 13 genes involved in carbon metabolism， sulfur metabolism， and nitrogen metabolism， respectively. Conclusion Highly efficient β-lactam antibiotic-degrading bacteria existed in seawater. Whole genome analysis revealed the possible degradation mechanism and bioremediation potential of ceftriaxone sodium-degrading bacterium DA02， providing a potential effective method for repairing β-lactam antibiotic pollution in the environment.

Key words: biodegradation; seawater; β-lactam antibiotics; whole genome

中图分类号:

S963.21^＋1

赵一宁, 潘瑞雪, 龙赟而, 林倩倩, 伍诗韵, 彭金菊, 马驿. 海洋源β-内酰胺类抗生素降解菌的分离鉴定及全基因组测序分析[J]. 中国畜牧兽医, 2026, 53(2): 984-994.

ZHAO Yining, PAN Ruixue, LONG Yuner, LIN Qianqian, WU Shiyun, PENG Jinju, MA Yi. Isolation， Identification and Whole Genome Sequencing Analysis of Marine-derived β-lactam Antibiotic-degrading Bacteria[J]. China Animal Husbandry & Veterinary Medicine, 2026, 53(2): 984-994.

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菌株 Strains	青霉素钾 Penicillin potassium	阿莫西林 Amoxicillin	头孢曲松钠 Ceftriaxone sodium
海水细菌Seawater bacteria	29.77±5.70	21.32±1.53	56.56±5.86
DP01	15.90±1.27	1.40±0.66	1.02±1.23
DP02	43.21±4.97	23.36±0.64	3.39±0.71
DP03	7.51±1.12	2.00±0.67	100.00±0.00
DP04	10.73±1.01	0.53±0.02	1.27±0.74
DP05	32.22±1.14	0.43±0.60	1.86±1.45
DP06	4.83±1.32	0.68±0.45	2.50±1.26
DP07	56.37±4.81	25.54±0.67	0.20±0.25
DA01	32.35±0.57	6.24±0.61	53.17±2.79
DA02	3.13±1.26	11.35±2.13	100.00±0.00
DA03	90.89±2.05	21.78±1.17	98.83±2.02
DA04	10.97±0.97	4.59±0.40	1.11±0.31
DA05	7.12±0.96	2.00±1.15	2.57±2.47
DA06	99.17±0.73	13.53±0.61	12.47±0.82
DT01	77.90±4.38	20.56±2.61	52.76±3.01
DT02	57.63±5.15	20.84±2.92	49.93±4.09
DT03	50.32±6.13	15.98±0.63	48.83±3.73
DT04	10.05±2.56	18.55±1.05	51.33±2.26
DT05	5.01±1.11	5.08±0.34	13.63±0.64