原生质体融合制备三效工程菌HL及其溶藻机理

doi:10.16085/j.issn.1000-6613.2021-2167

摘要/Abstract

摘要：

为从源头上治理蓝藻问题，对本文作者课题组筛选出的高效溶藻菌R1和脱氮聚磷菌B8进行原生质体融合后得到5株融合菌。以蓝藻优势藻种——铜绿微囊藻为受试对象，以溶藻率、脱氮率、除磷率作为评判标准，优选出其中1株兼备溶藻、脱氮、除磷菌株（命名为HL），运用底物抑制Haldane模型构建菌株降解铜绿微囊藻动力学模型。不同方式处理菌液来判定菌株的溶藻作用组分，并用扫描电镜观察其溶藻效果，辅以三维荧光、傅里叶红外初步探寻其溶藻机理。试验结果表明：菌株HL具有高效溶藻、稳定脱氮除磷能力；在菌藻共生环境中，对不同浓度铜绿微囊藻的降解过程符合Haldane方程，其最大比生长速率 $μ m$ 为1.046d^-1，半饱和常数 $K s$ 为896.92mg/m³，抑制常数K_I为1568.95mg/m³，这表明菌株HL对铜绿微囊藻具有较好的耐受与降解能力；菌株HL通过分泌芳香族氨基酸等活性物质，对藻细胞壁破坏，从而导致藻细胞破碎死亡。

关键词: 原生质体融合, 溶藻菌, 铜绿微囊藻, 动力学模型

Abstract:

In order to fundamentally control cyanobacteria, five strains of fusion bacteria were obtained by fusing protoplasm of the highly-efficiency Algicidal bacteria R1 and the denitrification and phosphorus accumulation bacterium B8 screened by research group. Taking Microcystis aeruginosa, the dominant algae species of cyanobacteria, as the test object, one of the strains (named HL) was optimized by the algae dissolution, nitrogen and phosphorus removal, and the kinetic model of degradation of Microcystis aeruginosa was constructed by using the substrate inhibition Haldane model. The algicidal effect was determined by treating the bacterial solution in different ways, and the algicidal effect was observed by scanning electron microscopy (SEM), supplemented by three-dimensional fluorescence and infrared spectroscopy to explore the mechanism of algae dissolution. The experimental results show that strain HL has the efficient algicidal ability, stable denitrification and phosphorus removal, and the degradation process of different concentrations of Microcystis aeruginosa in the symbiotic environment is in accordance with the Haldane equation, with the maximum specific growth rate ( $μ m$ ) of 1.046d^-1, half-saturation constant ( $K s$ ) of 896.92mg/m³and inhibition constant (K_I) of 1568.95mg/m³, which indicates that strain HL has good tolerance and degradation ability to Microcystis aeruginosa and damages algae cells wall by secreting active substances such as aromatic amino acids, leading to the fragmentation and death of algae cells.

Key words: protoplast fusion, Algicidal bacteria, Microcystis aeruginosa, dynamic model

中图分类号:

X522.1

黄金杰, 毛林强, 张文艺. 原生质体融合制备三效工程菌HL及其溶藻机理[J]. 化工进展, 2022, 41(8): 4464-4472.

HUANG Jinjie, MAO Linqiang, ZHANG Wenyi. Protoplast fusion preparation of three-effect engineered bacteria HL and its mechanism of algae dissolution[J]. Chemical Industry and Engineering Progress, 2022, 41(8): 4464-4472.

图/表 9

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荧光区域	代表物质类型	激发波长/nm	发射波长/nm
区域Ⅰ	酪氨酸等芳香族蛋白	220~250	280~330
区域Ⅱ	色氨酸等芳香族蛋白	220~250	330~380
区域Ⅲ	富里酸类物质	220~250	380~550
区域Ⅳ	微生物溶解性降解产物	250~400	280~380
区域Ⅴ	腐殖酸有机物	250~400	380~550

荧光区域	代表物质类型	激发波长/nm	发射波长/nm
区域Ⅰ	酪氨酸等芳香族蛋白	220~250	280~330
区域Ⅱ	色氨酸等芳香族蛋白	220~250	330~380
区域Ⅲ	富里酸类物质	220~250	380~550
区域Ⅳ	微生物溶解性降解产物	250~400	280~380
区域Ⅴ	腐殖酸有机物	250~400	380~550

菌株编号	大小/mm	形状	颜色	溶藻率/%	脱氮率/%	除磷率/%
1^#	3~4	圆形	白	78.15±0.35	41.23±2.97	45.51±2.11
2^#	1~2	圆形	白	67.10±2.33	27.12±1.63	29.95±1.26
3^#	3~4	圆形	乳白	77.26±1.25	38.27±2.64	14.29±2.75
4^#	1~2	圆形	透白	69.34±1.17	30.72±1.15	22.88±3.21
5^#	1~2	圆形	黄	81.61±1.94	58.64±1.07	61.78±1.32

菌株编号	大小/mm	形状	颜色	溶藻率/%	脱氮率/%	除磷率/%
1^#	3~4	圆形	白	78.15±0.35	41.23±2.97	45.51±2.11
2^#	1~2	圆形	白	67.10±2.33	27.12±1.63	29.95±1.26
3^#	3~4	圆形	乳白	77.26±1.25	38.27±2.64	14.29±2.75
4^#	1~2	圆形	透白	69.34±1.17	30.72±1.15	22.88±3.21
5^#	1~2	圆形	黄	81.61±1.94	58.64±1.07	61.78±1.32

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