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苏云金芽孢杆菌培养过程中的芽孢吡啶二羧酸检测

Endospore dipicolinic acid detection during Bacillus thuringiensis culture.

作者信息

Navarro A K, Peña A, Pérez-Guevara F

机构信息

Department of Molecular Genetics, Institute for Cellular Physiology, Universidad Nacional Autónoma de México, Mexico City, Mexico.

出版信息

Lett Appl Microbiol. 2008 Feb;46(2):166-70. doi: 10.1111/j.1472-765X.2007.02277.x. Epub 2007 Dec 7.

Abstract

AIMS

The aim of this work was to detect Bacillus thuringiensis endospore production during fermentation under conditions hindering endospore detection, i.e. in a complex undefined industrial medium with a high content of solids in suspension.

METHODS AND RESULTS

Bacterial endospore production was measured using the photoluminescence of dipicolinate (DPA) with Tb3+. The high temperature and pressure of a conventional autoclave was used to release DPA from the endospores. The endospore was obtained from B. thuringiensis var. kurstaki HD-73 fermentations in industrial-type media with 25.1 and 54.1 g l(-1) glucose, 4.4 and 35.3 g l(-1) soybean meal, 5.8 g l(-1) yeast extract, 9.2 g l(-1) corn steep solids and mineral salts.

CONCLUSIONS

In this study, we successfully determined the DPA concentrations during the culture of B. thuringiensis in high-concentration soybean meal media. A good correlation was found between microscope endospore counting and DPA measurements in the cultures.

SIGNIFICANCE AND IMPACT OF THE STUDY

Because of synergy between Cry protein and endospore in B. thuringiensis bioinsecticides formulation, it is important to be able to determine endospore development during B. thuringiensis industrial-type fermentation, in order to ascertain the beginning of sporulation.

摘要

目的

本研究旨在检测苏云金芽孢杆菌在不利于芽孢检测的发酵条件下,即在含有高悬浮固体含量的复杂未定义工业培养基中的芽孢产生情况。

方法与结果

使用Tb3 +与吡啶二羧酸(DPA)的光致发光来测量细菌芽孢的产生。利用传统高压灭菌器的高温高压从芽孢中释放DPA。芽孢取自苏云金芽孢杆菌库斯塔克变种HD - 73在工业型培养基中的发酵产物,该培养基含有25.1和54.1 g l(-1)葡萄糖、4.4和35.3 g l(-1)豆粕、5.8 g l(-1)酵母提取物、9.2 g l(-1)玉米浆干粉和矿物盐。

结论

在本研究中,我们成功测定了苏云金芽孢杆菌在高浓度豆粕培养基培养过程中的DPA浓度。在培养物中显微镜下芽孢计数与DPA测量之间发现了良好的相关性。

研究的意义与影响

由于苏云金芽孢杆菌生物杀虫剂制剂中Cry蛋白与芽孢之间的协同作用,能够在苏云金芽孢杆菌工业型发酵过程中确定芽孢发育情况以确定芽孢形成开始时间非常重要。

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