检测粘质沙雷氏菌 PRC-5 产生的几丁质酶 ChiA，使用抗 PrGV-几丁质酶。
Detection of chitinase ChiA produced by Serratia marcescens PRC-5, using anti-PrGV-chitinase.
机构信息
Division of Applied Bioscience and Biotechnology, Institute of Environmentally Friendly Agriculture (IEFA), Chonnam National University, Gwangju 500-757, Republic of Korea.
出版信息
Carbohydr Polym. 2013 Feb 15;92(2):2276-81. doi: 10.1016/j.carbpol.2012.12.018. Epub 2012 Dec 19.
In this study, a bacterium Serratia marcescens PRC-5 that displayed strong chitinolytic activity on 0.5% colloidal chitin-containing agar medium was isolated from soil. The chitinase activity increased rapidly with a maximum level (6.14 U/mL) on 4 days of incubation with swollen chitin (pH 5.0). Three active bands of chitinase isozymes were observed (53, 44, and 34 kDa) on SDS-PAGE gel and there pI values ranged from pI 5.4 to 5.8 on 2D gels. The chitinase from the PRC-5 strain was also able to produce GlcNAc monomers on TLC plates. The chitinase of PRC-5 inhibited the mycelial growth of Rhizoctonia solani KACC40111, which indicates that it could be used as a biocontrol agent for phytophathogens. The chitinase isozyme N1, which had a molecular weight of 62 kDa, was transferred from a native and SDS-PAGE gel onto an immunoblot and was probed using an anti-PrGV-chitinase.
从土壤中分离到一株能在含 0.5%胶体几丁质琼脂培养基上强烈分解几丁质的粘质沙雷氏菌（Serratia marcescens）PRC-5。当用膨胀几丁质（pH5.0）孵育 4 天时，几丁质酶活性迅速增加，达到最大水平（6.14U/mL）。在 SDS-PAGE 凝胶上观察到三个活性几丁质酶同工酶带（53、44 和 34kDa），在 2D 凝胶上的等电点（pI）值范围为 pI5.4 至 5.8。PRC-5 菌株的几丁质酶也能在 TLC 板上产生 GlcNAc 单体。PRC-5 几丁质酶能抑制立枯丝核菌（Rhizoctonia solani KACC40111）的菌丝生长，这表明它可以用作植物病原菌的生物防治剂。分子量为 62kDa 的几丁质酶同工酶 N1 从天然和 SDS-PAGE 凝胶转移到免疫印迹上，并使用抗-PrGV-几丁质酶进行探测。
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