分选特殊产物微生物的喇曼光谱法

以微生物油脂和类胡萝卜素为目标产物,通过对比目标产物与普通酵母的喇曼光谱,选择喇曼光谱的1 440 cm－1谱带与1 602 cm－1谱带的信号强度比值I1440/I1602作为油脂的特征标记,选择I1157/I1654或者I1520/I1654作为类胡萝卜素的标记,采用780 nm的激光俘获、收集酵母细胞的喇曼光谱,编写VB计算机程序,实时在线识别,通过光镊的操控,筛选产类胡萝卜素酵母和油脂酵母.经细胞培养和喇曼光谱检测验证了筛选结果.为筛选有经济价值的微生物提供了一种简单快捷的创新方法.

orting Economic Microorganisms by Using Optical Manipulation and Raman Microspectroscopy

A novel techique of sorting yeast cell based on Raman characteritics of target products synthesized by microorganisms was developed by using Raman microspectroscopy coupled with optical manipulatation.To sort oleaginous yeast and carotenoid-producing yeast,the plant oil and β-carotene were chosen as the target products and their Raman characteritics,the intensity ratios of 1 440 cm－1 and 1 602 cm－1 band (i.e.I1440/I1602) for oil,and I1157/I1654 or I1520/I1654 for carotenoid,were extracted and marked as target cells.Single yeast cell was captured and manipulated by using a 780 nm laser,and the Raman spectra of trapping cell was acquired simultaneously.A real-time computer distinguishing program was used to assist the sorting.The distinguished cells were then extracted and validated by following culture and Raman detection.This work provides a new method for screening microorganisms synthesize products with economic value.