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Chitinolytic proteins secreted by Cellulosimicrobium sp. NTK2.
FEMS Microbiology Letters ( IF 2.1 ) Pub Date : 2020-04-01 , DOI: 10.1093/femsle/fnaa055
Daisuke Niki 1 , Akari Higashitani 1 , Haruki Osada 2 , Tomohiro Bito 2 , Katsuhiko Shimizu 3 , Jiro Arima 2
Affiliation  

Cellulosimicrobium sp. NTK2 (NTK2 strain) was isolated as a chitinolytic bacterium from mature compost derived from chitinous waste. The growth of the NTK2 strain was enhanced by supplementation of the culture medium with 2% crystalline chitin. Approximately 70% of the supplemented crystalline chitin was degraded during cultivation. Whole genome analysis of the NTK2 strain identified eight chitinases and two chitin-binding proteins. The NTK2 strain secreted two bacterial extracellular solute-binding proteins, three family 18 glycosyl hydrolases and one lytic polysaccharide monooxygenase specifically in the presence of crystalline chitin. A chitinolytic enzyme with a molecular mass of 29 kDa on SDS-PAGE under native conditions was also secreted. This chitinolytic enzyme exhibited the largest band upon zymography but could not be identified. In an attempt to identify all the chitinases secreted by the NTK2 strain, we expressed recombinant versions of the proteins exhibiting chitinolytic activity in Escherichia coli. Our results suggest that the 29 kDa protein belonging to family 19 glycosyl hydrolase was expressed specifically in the presence of 2% crystalline chitin.

中文翻译:

Cellulosimicrobium sp. 分泌的几丁质分解蛋白。NTK2。

Cellulosimicrobium sp。NTK2(NTK2 菌株)作为几丁质分解细菌从来自几丁质废物的成熟堆肥中分离出来。NTK2 菌株的生长通过向培养基中添加 2% 结晶几丁质来增强。大约 70% 的补充结晶甲壳质在培养过程中被降解。NTK2 菌株的全基因组分析鉴定了八种几丁质酶和两种几丁质结合蛋白。NTK2 菌株在结晶几丁质存在下特异性分泌两种细菌细胞外溶质结合蛋白、三种家族 18 糖基水解酶和一种裂解多糖单加氧酶。在天然条件下,SDS-PAGE 上分子量为 29 kDa 的几丁质分解酶也被分泌。这种几丁质分解酶在酶谱中表现出最大的条带,但无法识别。为了鉴定 NTK2 菌株分泌的所有几丁质酶,我们表达了在大肠杆菌中表现出几丁质分解活性的蛋白质的重组版本。我们的结果表明,属于家族 19 糖基水解酶的 29 kDa 蛋白在 2% 结晶几丁质存在下特异性表达。
更新日期:2020-04-02
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