Synthetic Biology Journal ›› 2023, Vol. 4 ›› Issue (5): 980-999.DOI: 10.12211/2096-8280.2023-018
• Invited Review • Previous Articles Next Articles
Huan LIU1,2,3, Qiu CUI1,2,3
Received:
2023-03-01
Revised:
2023-07-27
Online:
2023-11-15
Published:
2023-10-31
Contact:
Qiu CUI
刘欢1,2,3, 崔球1,2,3
通讯作者:
崔球
作者简介:
基金资助:
CLC Number:
Huan LIU, Qiu CUI. Advances and applications of ambient ionization mass spectrometry in screening of microbial strains[J]. Synthetic Biology Journal, 2023, 4(5): 980-999.
刘欢, 崔球. 原位电离质谱技术在微生物菌株筛选中的应用进展[J]. 合成生物学, 2023, 4(5): 980-999.
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URL: https://synbioj.cip.com.cn/EN/10.12211/2096-8280.2023-018
电离技术 | 电离机理 | 应用场景 | 应用案例 |
---|---|---|---|
DESI | 电喷雾电离 气动辅助 | 细胞液干燥 | 亚种分化:大肠杆菌,鼠伤寒沙门氏菌菌株[ |
菌落 | 菌株表征:枯草芽孢杆菌[ | ||
微生物相互作用:大肠杆菌和黄色黏球菌[ | |||
表型分析:大肠杆菌[ | |||
nano-DESI | 电喷雾电离 | 菌落 | 代谢分析:希瓦氏菌、枯草芽孢杆菌、链霉菌[ |
分子网络:枯草芽孢杆菌,链霉菌,分枝杆菌和铜绿假单胞菌[ | |||
PSI | 电喷雾电离 纸基 | 菌落置于纸基 | 细菌区分:革兰氏阳性菌和革兰氏阴性菌的8个属16个物种[ |
藻液置于纸基 | 微藻表征:小球藻和微拟球藻[ | ||
MISI | 电喷雾电离 多级纸基 | 菌置于纸基 | 细菌鉴别:枯草芽孢杆菌、大肠杆菌和恶臭假单胞菌[ |
TD-ESI | 热解吸-电喷雾电离 | 菌落 | 细菌区分:5种革兰氏阳性菌和5种革兰氏阴性菌[ |
EASI | 声波电离 | 细胞液 | 膜脂质分析:蓝藻[ |
ITSME | 电喷雾电离 | 细胞脂滴 | 油脂检测:HepG2细胞[ |
Sciex Echo-MS | 电喷雾电离 | 细胞上清液或粗细胞提取液 | 酶筛选:二酰基甘油酰基转移酶2抑制剂[ |
微流体-ESI 微流体-MALDI | 电喷雾电离 激光电离 | 纳升液滴 纳升液滴 | 酶促反应:转氨酶ATA-117[ 酶分泌分析:酵母细胞中植酸酶[ |
LA-ESI | 激光烧蚀电喷雾电离 | 菌落 | 生长、代谢和抗生素抑制研究的分子成像:枯草芽孢杆菌和大肠杆菌[ |
表征:蓝藻[ | |||
微生物周转率:莱茵衣藻[ | |||
LA-REI | 激光蒸发电离 | 菌落 | 菌株分类:细菌和酵母[ |
MALDESI | 激光解吸电喷雾电离 | 细胞分散于载玻片上 | 脂质组学分析:HeLa细胞[ |
AP-LSI | 激光解吸电离 | 细胞液与基质共结晶 | 细菌区分:12个属的21种食源性细菌[ |
AP-MALDI | 激光解吸电离 | 菌落 | 成像:球状芽孢杆菌[ |
DART | 等离子体 | 全细胞悬浮液 | 细菌鉴定:化脓链球菌、大肠杆菌、γ-射线贝氏考克斯菌[ |
DBDI | 等离子体 | 细胞 | 化合物检测:洋葱细胞和PANC-1细胞[ |
LTP | 等离子体 | 细胞液沉积载玻片上 | 细菌区分:16种细菌的脂肪酸乙酯检测[ |
Table 1 AI techniques and their application examples in microbial detection
电离技术 | 电离机理 | 应用场景 | 应用案例 |
---|---|---|---|
DESI | 电喷雾电离 气动辅助 | 细胞液干燥 | 亚种分化:大肠杆菌,鼠伤寒沙门氏菌菌株[ |
菌落 | 菌株表征:枯草芽孢杆菌[ | ||
微生物相互作用:大肠杆菌和黄色黏球菌[ | |||
表型分析:大肠杆菌[ | |||
nano-DESI | 电喷雾电离 | 菌落 | 代谢分析:希瓦氏菌、枯草芽孢杆菌、链霉菌[ |
分子网络:枯草芽孢杆菌,链霉菌,分枝杆菌和铜绿假单胞菌[ | |||
PSI | 电喷雾电离 纸基 | 菌落置于纸基 | 细菌区分:革兰氏阳性菌和革兰氏阴性菌的8个属16个物种[ |
藻液置于纸基 | 微藻表征:小球藻和微拟球藻[ | ||
MISI | 电喷雾电离 多级纸基 | 菌置于纸基 | 细菌鉴别:枯草芽孢杆菌、大肠杆菌和恶臭假单胞菌[ |
TD-ESI | 热解吸-电喷雾电离 | 菌落 | 细菌区分:5种革兰氏阳性菌和5种革兰氏阴性菌[ |
EASI | 声波电离 | 细胞液 | 膜脂质分析:蓝藻[ |
ITSME | 电喷雾电离 | 细胞脂滴 | 油脂检测:HepG2细胞[ |
Sciex Echo-MS | 电喷雾电离 | 细胞上清液或粗细胞提取液 | 酶筛选:二酰基甘油酰基转移酶2抑制剂[ |
微流体-ESI 微流体-MALDI | 电喷雾电离 激光电离 | 纳升液滴 纳升液滴 | 酶促反应:转氨酶ATA-117[ 酶分泌分析:酵母细胞中植酸酶[ |
LA-ESI | 激光烧蚀电喷雾电离 | 菌落 | 生长、代谢和抗生素抑制研究的分子成像:枯草芽孢杆菌和大肠杆菌[ |
表征:蓝藻[ | |||
微生物周转率:莱茵衣藻[ | |||
LA-REI | 激光蒸发电离 | 菌落 | 菌株分类:细菌和酵母[ |
MALDESI | 激光解吸电喷雾电离 | 细胞分散于载玻片上 | 脂质组学分析:HeLa细胞[ |
AP-LSI | 激光解吸电离 | 细胞液与基质共结晶 | 细菌区分:12个属的21种食源性细菌[ |
AP-MALDI | 激光解吸电离 | 菌落 | 成像:球状芽孢杆菌[ |
DART | 等离子体 | 全细胞悬浮液 | 细菌鉴定:化脓链球菌、大肠杆菌、γ-射线贝氏考克斯菌[ |
DBDI | 等离子体 | 细胞 | 化合物检测:洋葱细胞和PANC-1细胞[ |
LTP | 等离子体 | 细胞液沉积载玻片上 | 细菌区分:16种细菌的脂肪酸乙酯检测[ |
电离技术 | 优劣势及适用场景 | 单个菌样 分析时间 | 空间分辨率 | 应用案例 |
---|---|---|---|---|
MALDI | 优势:方法成熟、样品制备简单、高通量 劣势:真空取样,限制不稳定或挥发性小分子的检测 适用场景:菌落 | 约2 s | 中链脂肪酸合成酶突变体库筛选:酿酒酵母 (m/z 730/758峰比值>1)[ 脂肪酸合成酶突变文库筛选:酿酒酵母(脂肪酸酰基片段C16:1/C18:1比值为18.1±1.1)、大肠杆菌(脂肪酸酰基片段C16:1/C16比值为1.11±0.04)和荧光假单胞菌(C16:1/C16比值为0.45±0.06)[ | |
1~2.5 s | 多步酶促反应:产肽类抗生素类似物的大肠杆菌和产鼠李糖脂的大肠杆菌[ | |||
5 s | 环二肽合酶定向进化:产二酮哌嗪的大肠杆菌[ | |||
约1 s | 120 μm× 120 μm | 菌落成像:铜绿假单胞菌、枯草芽孢杆菌和金黄色葡萄球菌[ | ||
约1 s | 250 μm | 菌落生物膜成像:枯草芽孢杆菌[ | ||
DESI | 优势:常压敞开条件下取样,原位、实时、高通量,适于细胞外泌代谢物和细胞膜相关分子 劣势:难以直接提供细胞内代谢物 适用场景:菌落、完整微生物细胞 | 10 s | 100 μm | 生物催化剂筛选:表达氨裂解酶和P450单加氧酶的大肠杆菌(L-苯丙氨酸检测限为>50 nmol/mm2)[ |
约7 s | 50 μm× 250 μm | 菌落成像:生产过量游离脂肪酸的大肠杆菌[ | ||
Nano-DESI | 同DESI | 1 mm × 1 mm | 质谱成像:希瓦氏菌、枯草芽孢杆菌和链霉菌[ | |
20 s | 约200 μm | 菌落质谱分子网络:枯草芽孢杆菌、链霉菌、分枝杆菌和铜绿假单胞菌[ | ||
LA-REI | 优势:常压敞开条件下取样,原位、实时、高通量、高分辨 劣势:限制细胞内代谢物直接分析 适用场景:菌落 | 8 s | 约500 μm | 微生物菌种分类:大肠杆菌等25种微生物[ |
10 s | 0.8 cm2 | 突变文库筛选:产紫色素和白桦酸的酿酒酵母(白桦酸线性范围2~2×10-5 g/L)[ | ||
LAESI | 优势:常压敞开条件下取样,原位、实时、高通量、高分辨 劣势:限制低极性化合物的分析 适用场景:菌落 | 150 μm | 菌落成像:枯草芽孢杆菌和大肠杆菌[ |
Table 2 Main AI techniques and their application examples in microbial strain screening and imaging
电离技术 | 优劣势及适用场景 | 单个菌样 分析时间 | 空间分辨率 | 应用案例 |
---|---|---|---|---|
MALDI | 优势:方法成熟、样品制备简单、高通量 劣势:真空取样,限制不稳定或挥发性小分子的检测 适用场景:菌落 | 约2 s | 中链脂肪酸合成酶突变体库筛选:酿酒酵母 (m/z 730/758峰比值>1)[ 脂肪酸合成酶突变文库筛选:酿酒酵母(脂肪酸酰基片段C16:1/C18:1比值为18.1±1.1)、大肠杆菌(脂肪酸酰基片段C16:1/C16比值为1.11±0.04)和荧光假单胞菌(C16:1/C16比值为0.45±0.06)[ | |
1~2.5 s | 多步酶促反应:产肽类抗生素类似物的大肠杆菌和产鼠李糖脂的大肠杆菌[ | |||
5 s | 环二肽合酶定向进化:产二酮哌嗪的大肠杆菌[ | |||
约1 s | 120 μm× 120 μm | 菌落成像:铜绿假单胞菌、枯草芽孢杆菌和金黄色葡萄球菌[ | ||
约1 s | 250 μm | 菌落生物膜成像:枯草芽孢杆菌[ | ||
DESI | 优势:常压敞开条件下取样,原位、实时、高通量,适于细胞外泌代谢物和细胞膜相关分子 劣势:难以直接提供细胞内代谢物 适用场景:菌落、完整微生物细胞 | 10 s | 100 μm | 生物催化剂筛选:表达氨裂解酶和P450单加氧酶的大肠杆菌(L-苯丙氨酸检测限为>50 nmol/mm2)[ |
约7 s | 50 μm× 250 μm | 菌落成像:生产过量游离脂肪酸的大肠杆菌[ | ||
Nano-DESI | 同DESI | 1 mm × 1 mm | 质谱成像:希瓦氏菌、枯草芽孢杆菌和链霉菌[ | |
20 s | 约200 μm | 菌落质谱分子网络:枯草芽孢杆菌、链霉菌、分枝杆菌和铜绿假单胞菌[ | ||
LA-REI | 优势:常压敞开条件下取样,原位、实时、高通量、高分辨 劣势:限制细胞内代谢物直接分析 适用场景:菌落 | 8 s | 约500 μm | 微生物菌种分类:大肠杆菌等25种微生物[ |
10 s | 0.8 cm2 | 突变文库筛选:产紫色素和白桦酸的酿酒酵母(白桦酸线性范围2~2×10-5 g/L)[ | ||
LAESI | 优势:常压敞开条件下取样,原位、实时、高通量、高分辨 劣势:限制低极性化合物的分析 适用场景:菌落 | 150 μm | 菌落成像:枯草芽孢杆菌和大肠杆菌[ |
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