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粘质沙雷氏菌几丁质酶高产突变体。

Chitinase-overproducing mutant of Serratia marcescens.

作者信息

Reid J D, Ogrydziak D M

出版信息

Appl Environ Microbiol. 1981 Mar;41(3):664-9. doi: 10.1128/aem.41.3.664-669.1981.

Abstract

Genetic modification of Serratia marcescens QMB1466 was undertaken to isolated mutants which produce increased levels of chitinolytic activity. After mutagenesis with ultraviolet light, ethyl methane sulfonate or N-methyl-N'-nitro-N-nitrosoguanidine, 19,940 colonies were screened for production of enlarged zones of clearing (indicative of chitinase activity) on chitin-containing agar plates. Forty-four chitinase high producers were tested further in shake flask cultures. Mutant IMR-1E1 was isolated which, depending on medium composition, produced two to three times more than the wild type of the other components of the chitinolytic enzyme system--a factor involved in the hydrolysis of crystalline chitin and chitobiase. After induction by chitin, endochitinase and chitobiase activity appeared at similar times for both IMR-1E1 and QMB1466, suggesting possible coordinate control of these enzymes. The results are consistent with IMR-1E1 containing a regulatory mutation which increased production of the components of the chitinolytic enzyme system and/or with IMR-1E1 containing a tandem duplication of the chitinase genes. The high rate of reversion of IMR-1E1 to decreased levels of chitinase production suggests that the overproduction of chitinase by IMR-1E1 is due to a tandem gene duplication.

摘要

对粘质沙雷氏菌QMB1466进行基因改造,以分离出几丁质酶活性水平提高的突变体。在用紫外线、甲基磺酸乙酯或N-甲基-N'-硝基-N-亚硝基胍诱变后,在含几丁质的琼脂平板上筛选了19940个菌落,以检测其是否产生扩大的透明圈(表明具有几丁质酶活性)。对44个几丁质酶高产菌株在摇瓶培养中进行了进一步测试。分离出了突变体IMR-1E1,根据培养基组成,其产生的几丁质酶解酶系统的其他成分比野生型多两到三倍,该系统是一种参与结晶几丁质水解的因子,还有几丁二糖酶。用几丁质诱导后,IMR-1E1和QMB1466的内切几丁质酶和几丁二糖酶活性在相似时间出现,这表明这些酶可能受到协同调控。结果与IMR-1E1含有一个调控突变相符,该突变增加了几丁质酶解酶系统成分的产量,和/或与IMR-1E1含有几丁质酶基因的串联重复相符。IMR-1E1回复到几丁质酶产量降低水平的高频率表明,IMR-1E1几丁质酶的过量产生是由于串联基因重复。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/75db/243756/4498b929fcf9/aem00196-0119-a.jpg

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