蛋白质谷氨酰胺酶产生菌的分离筛选和鉴定
Isolation and identification of nine bacteria strains producing Protein- glutaminase
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摘要: 目的筛选出能够产生蛋白质谷氨酰胺酶（Protein-glutaminase,PG）的菌株,并对筛选出的菌株分类和鉴定。方法以carboxybenzoxy（Cbz）-Gln-Gly为唯一氮源,从来自全国各地采集到的726份土样中富集筛选能够产生蛋白质谷氨酰胺酶的菌株,分别从形态学、生理学和分子生物学方面对所筛选的菌株进行分类鉴定,并测定发酵上清的脱酰胺活性。结果共筛选到9株产PG酶的细菌,经鉴定,其中7株为产吲哚金黄杆菌（Chryseobacterium indologenes）,一株为解朊金黄杆菌（Chryseobacterium proteolyticum）,另外一株为粘金黄杆菌（Chryseobacterium.gleum）。结论分别发酵测定9株菌的PG酶活性,产吲哚金黄杆菌ZYF120413-7发酵酶活最高,为0.7168U/m L。粘金黄杆菌D4-1-1的产酶能力最低,其酶活为0.1029U/m L。本课题的研究成果扩大了PG酶产生菌株的来源,为后续研究打下了基础。Abstract: ObjectiveIsolating and screening the strains producing Protein- glutaminase and then making Identification of them.Methods726 soil samples from all over the country were collected,from which the strains producing Protein- glutaminase were enriched with Cbz- Gln- Gly as the sole nitrogen source by preliminary screening in plates.Then the isolates were identified in morphology,physiology and molecular biology.The enzyme activity of deamidation was also been detected.ResultsTotally 9 strains that could produce PG were isolated,of which 7 bacteria were identified as Chryseobacterium indologenes,one was identified as Chryseobacterium proteolyticum and another was identified as Chryseobacterium.gleum.ConclusionsThe 9 strains were fermented in aerobic,and the PG activity in fermentation supernatant was tested. The results shown that Chryseobacterium indologenes ZYF120413- 7 had the highest enzyme activity with 0.7168 U / m L. Chryseobacterium. gleum came the lowest with only 0.1029 U / m L.This study enlarged the source of the PG producing strains,lying a good foundation for the following research.
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