Mery Sameh Waheeb, W. Elkhatib, M. Yassien, Nadia A. Hassouna
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引用次数: 2
摘要
纤维素酶因其应用的多样性而被认为是有效的生物催化剂。通过在羧甲基纤维素琼脂培养基上培养,从埃及土壤样品中回收了165株产纤维素酶链霉菌。刚果红法筛选结果表明,各分离株纤维素酶活性不同。采用二硝基水杨酸法对分离株纤维素酶产率进行定量评价,结果表明分离株纤维素酶产率较高(透明区≤25 mm, n= 15)。所选菌株的纤维素酶产量为146.9 ~ 650.5 U/L。通过16S rRNA基因测序,在分子水平上鉴定出产纤维素酶最高的分离物(S11-6)。序列同源性分析证实该菌株与coelicoflavus链霉菌NBRC 15399具有亲缘关系。利用4千格(KGy)的伽马辐射进行基因操作,提高纤维素酶的产量。与野生型菌株相比,M1突变体纤维素酶产量增加了1.31倍。
Production of Cellulase by Soil Isolated Streptomyces sp.
Cellulases have been considered effective biocatalysts because of their diversity of applications. A total of 165 cellulase-producing Streptomyces isolates were recovered from Egyptian soil samples via cultivation on carboxymethyl cellulose agar medium. The results of screening by the Congo red method showed that the tested isolates revealed different levels of cellulase activity. The isolates showed a high level of cellulase production (clear zones ˃ 25 mm, n= 15) were submitted for quantitative evaluation of their cellulase productivities by using dinitrosalicylic acid assay. Cellulase production of these selected isolates ranged from 146.9 to 650.5 U/L. The highest cellulase-producing isolate (S11-6) was identified at the molecular level using 16S rRNA gene sequencing. The sequence homology analysis proved its proximity and relatedness to Streptomyces coelicoflavus strain NBRC 15399. Production improvement of cellulase was carried out by genetic manipulation using a dose of 4 Kilogray (KGy) of gamma radiation. M1 mutant showed 1.31 fold increments in cellulase production as compared to the wild-type strain of S. coelicoflavus.