Chitin binding proteins act synergistically with chitinases in Serratia proteamaculans 568.
Chitin binding proteins act synergistically with chitinases in Serratia proteamaculans 568.
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DOI：
10.1371/journal.pone.0036714
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Podile AR
中科院分区：
文献类型：
--
作者：
Purushotham P;Arun PV;Prakash JS;Podile AR
变形斑沙雷氏菌568的基因组序列显示存在三个家族33几丁质结合蛋白（CBPs）。异源表达并纯化了三种Sp CBP（Sp CBP 21、Sp CBP 28和Sp CBP 50）。Sp CBP 21和Sp CBP 50对β-甲壳素有较强的结合能力，而Sp CBP 28不与甲壳素和纤维素结合。Sp CBP 21和Sp CBP 50均能与四种几丁质酶产生协同作用。Proteamaculans 568（Sp ChiA、Sp ChiB、Sp ChiC和Sp ChiD）对α-和β-几丁质降解的影响，特别是在外部电子供体（还原型谷胱甘肽）存在下。Sp ChiD在α-甲壳素上最受益于Sp CBP 21或Sp CBP 50，而Sp ChiB和Sp ChiD在β-甲壳素上主要受益于这些Sp CBP。剂量反应研究表明，两种Sp CBP均表现出≥0.2 µM的协同作用。将不同比例的Sp CBP 21和Sp CBP 50两者添加到协同混合物中没有显著增加活性。高度保守的极性残基，对S. marcescens（Sm CBP 21），而Sp CBP 28只有一个这样的极性残基。Sp CBP 28不能与供试底物结合可能是由于不存在重要的极性残基。
Genome sequence of Serratia proteamaculans 568 revealed the presence of three family 33 chitin binding proteins (CBPs). The three Sp CBPs (Sp CBP21, Sp CBP28 and Sp CBP50) were heterologously expressed and purified. Sp CBP21 and Sp CBP50 showed binding preference to β-chitin, while Sp CBP28 did not bind to chitin and cellulose substrates. Both Sp CBP21 and Sp CBP50 were synergistic with four chitinases from S. proteamaculans 568 (Sp ChiA, Sp ChiB, Sp ChiC and Sp ChiD) in degradation of α- and β-chitin, especially in the presence of external electron donor (reduced glutathione). Sp ChiD benefited most from Sp CBP21 or Sp CBP50 on α-chitin, while Sp ChiB and Sp ChiD had major advantage with these Sp CBPs on β-chitin. Dose responsive studies indicated that both the Sp CBPs exhibit synergism ≥0.2 µM. The addition of both Sp CBP21 and Sp CBP50 in different ratios to a synergistic mixture did not significantly increase the activity. Highly conserved polar residues, important in binding and activity of CBP21 from S. marcescens (Sm CBP21), were present in Sp CBP21 and Sp CBP50, while Sp CBP28 had only one such polar residue. The inability of Sp CBP28 to bind to the test substrates could be attributed to the absence of important polar residues.
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影响因子：
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