一株产生物表面活性剂油脂降解菌的分离及其特性研究

doi:10.3969/j.issn.1004-1524.20240410

浙江农业学报 ›› 2025, Vol. 37 ›› Issue (7): 1501-1511.DOI: 10.3969/j.issn.1004-1524.20240410

一株产生物表面活性剂油脂降解菌的分离及其特性研究

施添青(), 赵卓群, 谢晓杰, 郑华宝()

浙江农林大学环境与资源学院,浙江省土壤污染生物修复重点实验室,浙江杭州 311300

收稿日期:2024-05-08 出版日期:2025-07-25 发布日期:2025-08-20
作者简介:施添青(1998—),女,贵州兴仁人,硕士研究生,研究方向为农林废弃物资源化。E-mail:s3557544891@163.com
通讯作者: ^*郑华宝,E-mail:zhenghuabao@zafu.edu.cn
基金资助:
浙江省重点研发计划(2021C03190);浙江省“三农九方”农业科技协作计划(2022SNJF077)

Isolation and characterization of a biosurfactant production and oil-degrading bacteria strain

SHI Tianqing(), ZHAO Zhuoqun, XIE Xiaojie, ZHENG Huabao()

Key Laboratory of Soil Contamination Bioremediation of Zhejiang Province, College of Environment and Resources, Zhejiang A&F University, Hangzhou 311300, China

Received:2024-05-08 Online:2025-07-25 Published:2025-08-20

摘要/Abstract

摘要： 该研究从餐厨垃圾的废弃烹饪油(WCO)中筛选获得一株产生物表面活性剂的油脂降解菌——铜绿假单胞菌(Pseudomonas aeruginosa)STQ-2菌株。对该菌株产生物表面活性剂的发酵条件进行优化,并利用傅里叶变换红外光谱、核磁共振氢谱和液相色谱-电喷雾电离-质谱对其产物的结构进行分析。结果显示,该菌的最佳反应条件是30 ℃、pH值7、WCO质量浓度25 g·L^-1、以氯化铵作为氮源。当以WCO为唯一碳源,在上述条件下培养7 d后,菌株STQ-2的生物表面活性剂的产量达到600 mg·L^-1,WCO降解率达到21.1%。这表明,铜绿假单胞菌STQ-2既能降解WCO,又能以WCO作为唯一碳源生产生物表面活性剂。经鉴定,该菌所产生物表面活性剂的主要成分是鼠李糖脂,在pH值6~12、NaCl质量浓度20~140 g·L^-1、4~121 ℃的条件下,该生物表面活性剂具有化学稳定性。

关键词: 生物表面活性剂, 鼠李糖脂, 铜绿假单胞菌(Pseudomonas aeruginosa), 废弃烹饪油

Abstract:

In this study, a biosurfactant production and oil-degrading bacteria strain, Pseudomonas aeruginosa STQ-2, was isolated from waste cooking oil (WCO) derived from kitchen waste. The fermentation conditions were optimized to enhance the surfactant production by this strain, with the structural analysis of the products conducted by using Fourier transform infrared spectroscopy, nuclear magnetic resonance hydrogen spectroscopy, and liquid chromatography-electrospray ionization-mass spectrometry. It was shown that the optimal reaction conditions for this strain were as follows: temperature of 30 ℃, pH value of 7, WCO mass concentration of 25 g·L^-1, and ammonium chloride as the nitrogen source. When WCO was utilized as the sole carbon source, the yield of biosurfactant produced by strain STQ-2 reached 600 mg·L^-1, and the WCO degradation rate was 21.1% after 7 days of incubation under these conditions. These results demonstrated that Pseudomonas aeruginosa STQ-2 was capable of degrading WCO while simultaneously producing biosurfactant utilizing WCO as its sole carbon source. Furthermore, it was identified that the primary component of the surfactant produced by this strain was rhamnolipid, and the biosurfactant exhibited chemical stability across a pH value range of 6 to 12, NaCl mass concentrations of 20 to 140 g·L^-1, and temperature ranging from 4 to 121 ℃.

Key words: biosurfactant, rhamnolipid, Pseudomonas aeruginosa, waste cooking oil

中图分类号:

X172

施添青, 赵卓群, 谢晓杰, 郑华宝. 一株产生物表面活性剂油脂降解菌的分离及其特性研究[J]. 浙江农业学报, 2025, 37(7): 1501-1511.

SHI Tianqing, ZHAO Zhuoqun, XIE Xiaojie, ZHENG Huabao. Isolation and characterization of a biosurfactant production and oil-degrading bacteria strain[J]. Acta Agriculturae Zhejiangensis, 2025, 37(7): 1501-1511.

图/表 5

图1 STQ-2的菌落形态(A),及在透射电镜(200 nm)(B)、扫描电镜(3.0 kV,×9 000)(C)下的形态和在CTAB蓝平板上的显色结果(D)

Fig.1 Colony morphology (A), observatino result under transmission electron microscopy (200 nm) (B), scanning electron microscopy (3.0 kV,×9 000) (C), and color rendering results on CTAB solid plate (D) of STQ-2 strain

图2 基于STQ-2菌株16S rRNA序列的系统发育树

Fig.2 Phylogenetic tree based on 16S rRNA sequence of STQ-2 strain

图3 菌株STQ-2的培养条件优化及其对废弃烹饪油(WCO)的降解效果

Fig.3 Optimization of culture conditions for STQ-2 strain and its degradation capacity on waste cooking oil (WCO)

图4 生物表面活性剂粗品的表征 A,傅里叶变换红外光谱(FTIR);B,核磁共振氢谱(1H NMR);C~E,液相色谱-电喷雾电离-质谱(LC-ESI-MS)。RT,保留时间。

Fig.4 Structural characterization of crude biosurfactant A, Fourier Transform infrared spectroscopy (FTIR); B,1H nuclear magnetic resonance spectra (1H NMR);C-E, Liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS).RT,Retention time.

图5 表面张力和D600值随培养时间的变化(A),及生物表面活性剂在不同温度、pH值和盐浓度下的稳定性(B~D) E24,乳化活性。

Fig.5 Variation of surface tension and D600 during cultivation (A), and stability of biosurfactants at different temperature, pH value and salt concentrations (B-D) E24, Emulsifying properties.

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一株产生物表面活性剂油脂降解菌的分离及其特性研究

Isolation and characterization of a biosurfactant production and oil-degrading bacteria strain

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